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Sample records for plasma ttr binding

  1. Levels and distributions of organic pollutants in subtidal sediments from the Loire estuary: Are there any relationships with TTR-binding activity?

    Science.gov (United States)

    Couderc, M.; Gandolfi, F.; Zalouk-Vergnoux, A.; Beyeler, N.; Malleret, L.; Ambidina, I.; Kamari, A.; Blanchet-Letrouvé, I.; Mouneyrac, C.; Hamers, T.; Poirier, L.

    2016-12-01

    The Loire estuary runs through important urban sites with shipping, industrial and agricultural activities, being the receptacle of diffusive pollutants comprising, a mixture of contaminants such as persistent organic pollutants (POPs). This work was set out to evaluate the occurrence of thyroid endocrine disruptors in sediments of this estuary. Sediments were collected in September 2012 and April 2013, in subtidal zones along the estuary. Targeted chemical analyses of five classes of pollutants, i.e. polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), polycyclic aromatic hydrocarbons (PAHs), alkylphenols (APs), and bisphenol A (BPA) were performed in sediment extracts. Extracts were further tested for their thyroid hormone (TH) disrupting potency to compete with TH for binding to its transporter protein transthyretin (TTR). The Haute-Indre site was characterized by a significant PAH contamination whereas Saint-Nazaire, Bellevue and Rezé would be particularly contaminated by PCBs. These observations could be linked to the different type of anthropogenic activities taking place close to these sites. Donges, Mindin and Paimboeuf were the sampling sites displaying the lowest contamination in PAHs, APs, PCBs and PBDEs. No inter-site difference could be observed for TTR-binding activity, which should be attributed to different compounds than the chemically analyzed compounds, as confirmed by PCA analyses. Furthermore, the TTR-binding potencies of the extracts were relatively low compared to data from literature. More investigations on the quantification of PCB and PBDE hydroxylated metabolites and other known endocrine disruptors such as pesticides or perfluorinated compounds could be considered, as well as bioassays highlighting other endocrine disrupting effects.

  2. Production TTR modeling and dynamic buckling analysis

    Institute of Scientific and Technical Information of China (English)

    Hugh Liu; John Wei; Edward Huang

    2013-01-01

    In a typical tension leg platform (TLP) design,the top tension factor (TTF),measuring the top tension of a top tensioned riser (TTR) relative to its submerged weight in water,is one of the most important design parameters that has to be specified properly.While a very small TTF may lead to excessive vortex induced vibration (ⅤⅣ),clashing issues and possible compression close to seafloor,an unnecessarily high TTF may translate into excessive riser cost and vessel payload,and even has impacts on the TLP sizing and design in general.In the process of a production TTR design,it is found that its outer casing can be subjected to compression in a worst-case scenario with some extreme metocean and hardware conditions.The present paper shows how finite element analysis (FEA) models using beam elements and two different software packages (Flexcom and ABAQUS) are constructed to simulate the TTR properly,and especially the pipe-in-pipe effects.An ABAQUS model with hybrid elements (beam elements globally + shell elements locally) can be used to investigate how the outer casing behaves under compression.It is shown for the specified TTR design,even with its outer casing being under some local compression in the worst-case scenario,dynamic buckling would not occur; therefore the TTR design is adequate.

  3. Stereoselective binding of chiral drugs to plasma proteins

    Institute of Scientific and Technical Information of China (English)

    Qi SHEN; Lu WANG; Hui ZHOU; Hui-di JIANG; Lu-shan YU; Su ZENG

    2013-01-01

    Chiral drugs show distinct biochemical and pharmacological behaviors in the human body.The binding of chiral drugs to plasma proteins usually exhibits stereoselectivity,which has a far-reaching influence on their pharmacological activities and pharmacokinetic profiles.In this review,the stereoselective binding of chiral drugs to human serum albumin (HSA),α1-acid glycoprotein (AGP)and lipoprotein,three most important proteins in human plasma,are detailed.Furthermore,the application of AGP variants and recombinant fragments of HSA for studying enantiomer binding properties is also discussed.Apart from the stereoselectivity of enantiomer-protein binding,enantiomer-enantiomer interactions that may induce allosteric effects are also described.Additionally,the techniques and methods used to determine drug-protein binding parameters are briefly reviewed.

  4. [Determination of plasma protein binding rate of arctiin and arctigenin with ultrafiltration].

    Science.gov (United States)

    Han, Xue-Ying; Wang, Wei; Tan, Ri-Qiu; Dou, De-Qiang

    2013-02-01

    To determine the plasma protein binding rate of arctiin and arctigenin. The ultrafiltration combined with HPLC was employed to determine the plasma protein binding rate of arctiin and arctigenin as well as rat plasma and healthy human plasma proteins. The plasma protein binding rate of arctiin with rat plasma at the concentrations of 64. 29, 32.14, 16.07 mg x L(-1) were (71.2 +/- 2.0)%, (73.4 +/- 0.61)%, (78.2 +/- 1.9)%, respectively; while the plasma protein binding rate of arctiin with healthy human plasma at the above concentrations were (64.8 +/- 3.1)%, (64.5 +/- 2.5)%, (77.5 +/- 1.7)%, respectively. The plasma protein binding rate of arctigenin with rat plasma at the concentrations of 77.42, 38.71, 19.36 mg x L(-1) were (96.7 +/- 0.41)%, (96.8 +/- 1.6)%, (97.3 +/- 0.46)%, respectively; while the plasma protein binding rate of arctigenin with normal human plasma at the above concentrations were (94.7 +/- 3.1)%, (96.8 +/- 1.6)%, (97.9 +/- 1.3)%, respectively. The binding rate of arctiin with rat plasma protein was moderate, which is slightly higher than the binding rate of arctiin with healthy human plasma protein. The plasma protein binding rates of arctigenin with both rat plasma and healthy human plasma are very high.

  5. Differential plasma protein binding to metal oxide nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Deng, Zhou J; Mortimer, Gysell; Minchin, Rodney F [School of Biomedical Sciences, University of Queensland, Brisbane, QLD 4072 (Australia); Schiller, Tara; Musumeci, Anthony; Martin, Darren, E-mail: r.minchin@uq.edu.a [Australian Institute for Bioengineering and Nanotechnology, University of Queensland, Brisbane, QLD 4072 (Australia)

    2009-11-11

    Nanoparticles rapidly interact with the proteins present in biological fluids, such as blood. The proteins that are adsorbed onto the surface potentially dictate the biokinetics of the nanomaterials and their fate in vivo. Using nanoparticles with different sizes and surface characteristics, studies have reported the effects of physicochemical properties on the composition of adsorbed plasma proteins. However, to date, few studies have been conducted focusing on the nanoparticles that are commonly exposed to the general public, such as the metal oxides. Using previously established ultracentrifugation approaches, two-dimensional gel electrophoresis and mass spectrometry, the current study investigated the binding of human plasma proteins to commercially available titanium dioxide, silicon dioxide and zinc oxide nanoparticles. We found that, despite these particles having similar surface charges in buffer, they bound different plasma proteins. For TiO{sub 2}, the shape of the nanoparticles was also an important determinant of protein binding. Agglomeration in water was observed for all of the nanoparticles and both TiO{sub 2} and ZnO further agglomerated in biological media. This led to an increase in the amount and number of different proteins bound to these nanoparticles. Proteins with important biological functions were identified, including immunoglobulins, lipoproteins, acute-phase proteins and proteins involved in complement pathways and coagulation. These results provide important insights into which human plasma proteins bind to particular metal oxide nanoparticles. Because protein absorption to nanoparticles may determine their interaction with cells and tissues in vivo, understanding how and why plasma proteins are adsorbed to these particles may be important for understanding their biological responses.

  6. QSAR Models for the Prediction of Plasma Protein Binding

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    Zeshan Amin

    2013-02-01

    Full Text Available Introduction: The prediction of plasma protein binding (ppb is of paramount importance in the pharmacokinetics characterization of drugs, as it causes significant changes in volume of distribution, clearance and drug half life. This study utilized Quantitative Structure – Activity Relationships (QSAR for the prediction of plasma protein binding. Methods: Protein binding values for 794 compounds were collated from literature. The data was partitioned into a training set of 662 compounds and an external validation set of 132 compounds. Physicochemical and molecular descriptors were calculated for each compound using ACD labs/logD, MOE (Chemical Computing Group and Symyx QSAR software packages. Several data mining tools were employed for the construction of models. These included stepwise regression analysis, Classification and Regression Trees (CART, Boosted trees and Random Forest. Results: Several predictive models were identified; however, one model in particular produced significantly superior prediction accuracy for the external validation set as measured using mean absolute error and correlation coefficient. The selected model was a boosted regression tree model which had the mean absolute error for training set of 13.25 and for validation set of 14.96. Conclusion: Plasma protein binding can be modeled using simple regression trees or multiple linear regressions with reasonable model accuracies. These interpretable models were able to identify the governing molecular factors for a high ppb that included hydrophobicity, van der Waals surface area parameters, and aromaticity. On the other hand, the more complicated ensemble method of boosted regression trees produced the most accurate ppb estimations for the external validation set.

  7. Nasal mucosa: a new site for tissue biopsy to diagnose hereditary TTR amyloidosis.

    Science.gov (United States)

    Munar-Qués, Miguel; Solé, Manel; Martínez-Nadal, Jacinto; Murcia-Sáiz, Antonio; Mas-Degano, José Manuel

    2014-11-07

    We report 2 carriers of the TTRV30M mutation and its plasmatic biochemical marker with clinical symptoms compatible with hereditary TTR amyloidosis. Based on our previously reported casual finding of amyloid TTR in nasal mucosa (2008), we requested biopsy of this tissue to search for amyloid with Congo red staining and TTR immunohistochemical analysis. The histological diagnosis was achieved by retrospective analysis of surgical sinonasal biopsy in the first patient and prospective biopsy of inferior nasal concha in the second. Large interstitial deposits of ATTR were observed in both cases. We suggest nasal mucosa as a suitable site for tissue biopsy in patients with suspected hereditary TTR amyloidosis. Copyright © 2014 Elsevier España, S.L.U. All rights reserved.

  8. Genotypic and phenotypic presentation of transthyretin-related familial amyloid polyneuropathy (TTR-FAP) in Turkey.

    Science.gov (United States)

    Durmuş-Tekçe, Hacer; Matur, Zeliha; Mert Atmaca, Murat; Poda, Mehves; Çakar, Arman; Hıdır Ulaş, Ümit; Oflazer-Serdaroğlu, Piraye; Deymeer, Feza; Parman, Yesim G

    2016-07-01

    Transthyretin-related familial amyloid polyneuropathy (TTR-FAP) is an autosomal dominant disorder caused by mutations of the transthyretin (TTR) gene. The mutant amyloidogenic transthyretin protein causes the systemic accumulation of amyloid fibrils that result in organ dysfunction. TTR-associated FAP is a progressive and fatal disease, if left untreated, and should be considered in the differential diagnosis of any person presenting with a progressive polyneuropathy, particularly with accompanying autonomic involvement. The clinical, electrophysiological, histopathological, and genetic characteristics of 17 patients from Turkey (5 female, 13 male) from nine families with polyneuropathy and mutations in TTR were evaluated. Sequence analysis of the TTR gene revealed five mutations (Val30Met, Glu89Gln, Gly53Glu, Glu54Gly and Gly47Glu). Mean age at disease onset was 40.4 ± 13.9 years (range 21-66 years). The most commonly reported initial complaint was paresthesia in the feet (asymmetric in three patients). Three patients (2 male) with the Glu89Gln mutation presented with carpal tunnel syndrome. Two patients with the Gly53Glu mutation showed episodes of dysarthria and hemiparesis, consistent with this genotype. Seven patients died during the period of follow-up as a result of systemic involvement. Our study suggests that a cohort of patients from Turkey with TTR-FAP exhibits clinical and genetic heterogeneity.

  9. Differential dissociation micromethod for the investigation of binding of metandrostenolone (Nerobol) to plasma proteins

    Energy Technology Data Exchange (ETDEWEB)

    Bojadzsieva, M.; Kocsar, L. (Orszagos Frederic Joliot-Curie Sugarbiologiai es Sugaregeszseguegyi Kutato Intezet, Budapest (Hungary)); Kremmer, T. (Orszagos Onkologiai Intezet, Budapest (Hungary))

    1985-01-01

    A micromethod was developed to determine the binding of anabolic steroids to plasma proteins. The new procedure combines precipitation with ammonium sulphate and differential dissociation. The binding parameters (association constant, specific binding capacity) are calculated on the basis of dissociation curves of sup(3)H-metandrostenolone from the precipitated sexual binding globuline.

  10. ENDEAVOUR: Phase 3 Multicenter Study of Revusiran (ALN-TTRSC) in Patients With Transthyretin (TTR) Mediated Familial Amyloidotic Cardiomyopathy (FAC)

    Science.gov (United States)

    2016-10-11

    Transthyretin (TTR) Mediated Familial Amyloidotic Cardiomyopathy (FAC); Amyloidosis, Hereditary; Amyloid Neuropathies, Familial; Amyloid Neuropathies; Amyloidosis, Hereditary, Transthyretin-Related; Familial Transthyretin Cardiac Amyloidosis

  11. Characterization of auxin-binding proteins from zucchini plasma membrane

    Science.gov (United States)

    Hicks, G. R.; Rice, M. S.; Lomax, T. L.

    1993-01-01

    We have previously identified two auxin-binding polypeptides in plasma membrane (PM) preparations from zucchini (Cucurbita pepo L.) (Hicks et al. 1989, Proc. Natl. Acad. Sci. USA 86, 4948-4952). These polypeptides have molecular weights of 40 kDa and 42 kDa and label specifically with the photoaffinity auxin analog 5-N3-7-3H-IAA (azido-IAA). Azido-IAA permits both the covalent and radioactive tagging of auxin-binding proteins and has allowed us to characterize further the 40-kDa and 42-kDa polypeptides, including the nature of their attachment to the PM, their relationship to each other, and their potential function. The azido-IAA-labeled polypeptides remain in the pelleted membrane fraction following high-salt and detergent washes, which indicates a tight and possibly integral association with the PM. Two-dimensional electrophoresis of partially purified azido-IAA-labeled protein demonstrates that, in addition to the major isoforms of the 40-kDa and 42-kDa polypeptides, which possess isoelectric points (pIs) of 8.2 and 7.2, respectively, several less abundant isoforms that display unique pIs are apparent at both molecular masses. Tryptic and chymotryptic digestion of the auxin-binding proteins indicates that the 40-kDa and 42-kDa polypeptides are closely related or are modifications of the same polypeptide. Phase extraction with the nonionic detergent Triton X-114 results in partitioning of the azido-IAA-labeled polypeptides into the aqueous (hydrophilic) phase. This apparently paradoxical behavior is also exhibited by certain integral membrane proteins that aggregate to form channels. The results of gel filtration indicate that the auxin-binding proteins do indeed aggregate strongly and that the polypeptides associate to form a dimer or multimeric complex in vivo. These characteristics are consistent with the hypothesis that the 40-kDa and 42-kDa polypeptides are subunits of a multimeric integral membrane protein which has an auxin-binding site, and which may

  12. MLKL Compromises Plasma Membrane Integrity by Binding to Phosphatidylinositol Phosphates

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    Yves Dondelinger

    2014-05-01

    Full Text Available Although mixed lineage kinase domain-like (MLKL protein has emerged as a specific and crucial protein for necroptosis induction, how MLKL transduces the death signal remains poorly understood. Here, we demonstrate that the full four-helical bundle domain (4HBD in the N-terminal region of MLKL is required and sufficient to induce its oligomerization and trigger cell death. Moreover, we found that a patch of positively charged amino acids on the surface of the 4HBD binds to phosphatidylinositol phosphates (PIPs and allows recruitment of MLKL to the plasma membrane. Importantly, we found that recombinant MLKL, but not a mutant lacking these positive charges, induces leakage of PIP-containing liposomes as potently as BAX, supporting a model in which MLKL induces necroptosis by directly permeabilizing the plasma membrane. Accordingly, we found that inhibiting the formation of PI(5P and PI(4,5P2 specifically inhibits tumor necrosis factor (TNF-mediated necroptosis but not apoptosis.

  13. Association of TTR polymorphisms with hippocampal atrophy in Alzheimer disease families.

    Science.gov (United States)

    Cuenco, Karen T; Friedland, Robert; Baldwin, Clinton T; Guo, Jianping; Vardarajan, Badri; Lunetta, Kathryn L; Cupples, L Adrienne; Green, Robert C; DeCarli, Charles; Farrer, Lindsay A

    2011-02-01

    In vitro and animal model studies suggest that transthyretin (TTR) inhibits the production of the amyloid β protein, a major contributor to Alzheimer disease (AD) pathogenesis. We evaluated the association of 16 TTR single nucleotide polymorphisms (SNPs) with AD risk in 158 African American and 469 Caucasian discordant sibships from the MIRAGE Study. There was no evidence for association of TTR with AD in either population sample. To examine the possibility that TTR SNPs affect specific components of the AD process, we tested association of these SNPs with four measures of neurodegeneration and cerebrovascular disease defined by magnetic resonance imaging (MRI) in a subset of 48 African American and 265 Caucasian sibships. Five of seven common SNPs and several haplotypes were significantly associated with hippocampal atrophy in the Caucasian sample. Two of these SNPs also showed marginal evidence for association in the African American sample. Results for the other MRI traits were unremarkable. This study highlights the potential value of neuroimaging endophenotypes as a tool for finding genes influencing AD pathogenesis.

  14. Inhibition of platelet (/sup 3/H)- imipramine binding by human plasma protein fractions

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    Strijewski, A.; Chudzik, J.; Tang, S.W.

    1988-01-01

    Inhibition of high-affinity (/sup 3/H)-imipramine binding to platelet membranes by human plasma fractions and isolated plasma proteins was investigated. Several plasma proteins were found to contribute to the observed apparent inhibition and this contribution was assessed in terms of inhibitor units. Alpha/sub 1/ acid glycoprotein, high density and low density lipoprotein, IgG and ..cap alpha../sub 1/-antitrypsin were identified as effective non-specific inhibitors. Alpha-1-acid glycoprotein was confirmed to be the most potent plasma protein inhibitor. Cohn fractions were evaluated for the presence of the postulated endocoid of (/sup 3/H)-imipramine binding site.

  15. Binding patterns of seminal plasma plasma proteins on bovine epididymal and ejaculated sperm membrane

    Directory of Open Access Journals (Sweden)

    C.E.A. Souza

    2011-06-01

    Full Text Available The present study was designed to investigate the topographical distribution of seminal plasma (SP proteins on epididymal and ejaculated bovine sperm. Using immunocytochemistry and confocal microscopy the binding patterns of bovine SP proteins BSP-A3, albumin, transferrin, prostaglandin D-synthase (PGDS and nucleobindin in ejaculated and cauda epididymal sperm from adult bulls were evaluated. Experiments were performed using sperm from 5 males. Data showed a positive signal, only detected for anti-PGDS, in the acrosomal cap of epididymal and ejaculated sperm. In ejaculated sperm, a very weak signal for nucleobindin 2 in the midpiece and equatorial regions was detected, using the anti-rat nucleobindin. BSP-A3 was detected on all sperm regions studied, with a more evidenced signal in acrosome and midpiece. However, no binding was detected for albumin or transferrin in neither epididymal nor ejaculated sperm. In conclusion, PGDS, BSP-A3 and nucleobindin interact directly with bovine sperm, with specific topographic distribution. These findings may add to the knowledge of how these proteins modulate sperm functions, thus providing fundamental support for studies designed to evaluate how they influence sperm functions.

  16. Automation of plasma protein binding assay using rapid equilibrium dialysis device and Tecan workstation.

    Science.gov (United States)

    Ye, Zhengqi; Zetterberg, Craig; Gao, Hong

    2017-03-14

    Binding of drug molecules to plasma proteins is an important parameter in assessing drug ADME properties. Plasma protein binding (PPB) assays are routinely performed during drug discovery and development. A fully automated PPB assay was developed using rapid equilibrium dialysis (RED) device and Tecan workstation coupled to an automated incubator. The PPB assay was carried out in unsealed RED plates which allowed the assay to be fully automated. The plasma pH was maintained at 7.4 during the 6-h dialysis under 2% CO2 condition. The samples were extracted with acetonitrile and analyzed by liquid chromatography tandem mass spectrometry. The percent bound results of 10 commercial drugs in plasma protein binding were very similar between the automated and manual assays, and were comparable to literature values. The automated assay increases laboratory productivity and is applicable to high-throughput screening of drug protein binding in drug discovery.

  17. Valproic acid: in vitro plasma protein binding and interaction with phenytoin.

    Science.gov (United States)

    Cramer, J A; Mattson, R H

    1979-01-01

    Because valproic acid (VPA) is highly bound to plasma protein, several variables affecting binding will significantly alter the quantity of free drug which is pharmacologically active. Therefore, total VPA plasma concentrations do not reflect the therapeutic strength of the drug in tissue. We have performed equilibrium dialysis and ultrafiltration studies of VPA binding to plasma protein. The converging data in these in vitro studies indicate a clinically significant alteration in the percent of free VPA when total drug concentration exceeds 80 micrograms/ml. Saturation of drug binding sites probably occurs in this range. At 20--60 micrograms/ml VPA there is 5% free drug, with a significant increase to 8% free at 80 micrograms/ml; free drug increases to over 20% at 145 micrograms/ml total VPA. Human plasma, which is low in albumin, has twice the quantity of free VPA as normal plasma (10 versus 5% free). The clinical evidence of interaction between VPA and phenytoin is confirmed in vitro by the increase in the free fraction of both drugs. VPA binding decreases by 3--6%, while phenytoin binding decreases 5--6% as both drugs reach high plasma concentrations. When appropriate, laboratory reports should be available defining concentration of free drug in plasma for optimal interpretation of drug concetrations relative to clinical effects.

  18. Early Diagnosis of Intestinal Ischemia Using Urinary and Plasma Fatty Acid Binding Proteins

    NARCIS (Netherlands)

    Thuijls, Geertje; van Wijck, Kim; Grootjans, Joep; Derikx, Joep P. M.; van Bijnen, Annemarie A.; Heineman, Erik; Dejong, Cornelis H. C.; Buurman, Wim A.; Poeze, Martijn

    2011-01-01

    Objective: This study aims at improving diagnosis of intestinal ischemia, by measuring plasma and urinary fatty acid binding protein (FABP) levels. Methods: Fifty consecutive patients suspected of intestinal ischemia were included and blood and urine were sampled at time of suspicion. Plasma and uri

  19. Inhibitors of serotonin reuptake and specific imipramine binding in human blood plasma

    Energy Technology Data Exchange (ETDEWEB)

    Brusov, O.S.; Fomenko, A.M.; Katasonov, A.B.; Lidemann, R.R.

    1985-12-01

    This paper describes a method of extraction of endogenous inhibitors of specific IMI binding and of 5-HT reuptake, from human blood plasma and the heterogeneity of these compounds is demonstrated. Specific binding was determined as the difference between binding of /sup 3/H-IMI in the absence and in the presence of 50 microM IMI. Under these conditions, specific binding amounted to 70-80% of total binding of /sup 3/H-IMI. It is shown that extract obtained from human blood contains a material which inhibits dose-dependently both 5-HT reuptake and specific binding of /sup 3/H-IMI. Gel-chromatography of extracts of human blood plasma on Biogel P-2 is also shown.

  20. Effect of anticonvulsants on plasma testosterone and sex hormone binding globulin levels.

    Science.gov (United States)

    Barragry, J M; Makin, H L; Trafford, D J; Scott, D F

    1978-01-01

    Plasma sex hormone binding globulin (SHBG) and testosterone levels were measured in 29 patients with epilepsy (16 men and 13 women), most of them on chronic therapy with anticonvulsant drugs. Sex hormone binding globulin concentrations were increased in both sexes and testosterone levels in male patients. It is postulated that anticonvulsants may induce hepatic synthesis of SHBG. PMID:569688

  1. A rapid and simple assay for growth hormone-binding protein activity in human plasma.

    Science.gov (United States)

    Baumann, G; Shaw, M A; Amburn, K

    1988-12-01

    The newly discovered circulating growth hormone binding proteins dictate a re-evaluation of the state of GH in plasma in health and disease as the binding proteins are known to affect GH metabolism and action. We describe a rapid and simple GH-binding assay that allows determination of free and complexed plasma GH, as well as GH-binding protein activity as an index of GH-binding protein levels, with relative ease. The method is based on incubation of plasma with 125I-GH and separation of bound from free GH on small DEAE-cellulose columns; it can be used on a large scale for routine determinations. The results obtained by this method are comparable to those obtained with the previously used slow and more cumbersome gel filtration technique. Initial data obtained in normal subjects and certain disease states show that the bound fraction of plasma GH is similar in men, women and children, is unaffected by pregnancy or acute infection, but is marginally decreased in liver cirrhosis. In acromegaly, binding protein activity also appears normal when allowance is made for partial saturation of the binding proteins by the high prevailing GH levels. The technique we describe should facilitate investigations of normal and abnormal regulation of the GH binding proteins.

  2. Relationship between brain serotonin transporter binding, plasma concentration and behavioural effect of selective serotonin reuptake inhibitors

    OpenAIRE

    2005-01-01

    The present study was undertaken to characterise the relationship between in vivo brain serotonin transporter (SERT) binding, plasma concentration and pharmacological effect of selective serotonin reuptake inhibitors (SSRIs) in mice. Oral administration of fluvoxamine, fluoxetine, paroxetine and sertraline at pharmacologically relevant doses exerted dose- and time-dependent binding activity of brain SERT as revealed by significant increases in KD for specific [3H]paroxetine binding, and the i...

  3. QSARs for Plasma Protein Binding: Source Data and Predictions

    Data.gov (United States)

    U.S. Environmental Protection Agency — The dataset has all of the information used to create and evaluate 3 independent QSAR models for the fraction of a chemical unbound by plasma protein (Fub) for...

  4. Rational use of plasma protein and tissue binding data in drug design.

    Science.gov (United States)

    Liu, Xingrong; Wright, Matthew; Hop, Cornelis E C A

    2014-10-23

    It is a commonly accepted assumption that only unbound drug molecules are available to interact with their targets. Therefore, one of the objectives in drug design is to optimize the compound structure to increase in vivo unbound drug concentration. In this review, theoretical analyses and experimental observations are presented to illustrate that low plasma protein binding does not necessarily lead to high in vivo unbound plasma concentration. Similarly, low brain tissue binding does not lead to high in vivo unbound brain tissue concentration. Instead, low intrinsic clearance leads to high in vivo unbound plasma concentration, and low efflux transport activity at the blood-brain barrier leads to high unbound brain concentration. Plasma protein and brain tissue binding are very important parameters in understanding pharmacokinetics, pharmacodynamics, and toxicities of drugs, but these parameters should not be targeted for optimization in drug design.

  5. Multiple sup 3 H-oxytocin binding sites in rat myometrial plasma membranes

    Energy Technology Data Exchange (ETDEWEB)

    Crankshaw, D.; Gaspar, V.; Pliska, V. (McMaster Univ., Hamilton, Ontario, (Canada))

    1990-01-01

    The affinity spectrum method has been used to analyse binding isotherms for {sup 3}H-oxytocin to rat myometrial plasma membranes. Three populations of binding sites with dissociation constants (Kd) of 0.6-1.5 x 10(-9), 0.4-1.0 x 10(-7) and 7 x 10(-6) mol/l were identified and their existence verified by cluster analysis based on similarities between Kd, binding capacity and Hill coefficient. When experimental values were compared to theoretical curves constructed using the estimated binding parameters, good fits were obtained. Binding parameters obtained by this method were not influenced by the presence of GTP gamma S (guanosine-5'-O-3-thiotriphosphate) in the incubation medium. The binding parameters agree reasonably well with those found in uterine cells, they support the existence of a medium affinity site and may allow for an explanation of some of the discrepancies between binding and response in this system.

  6. Sex steroid binding proteins in the plasma of hatchling Chelonia mydas.

    Science.gov (United States)

    Ikonomopoulou, M P; Ibrahim, K; Bradley, A J

    2008-09-01

    Sex steroid binding proteins were identified in hatchling female and male Chelonia mydas by dialysis and steady-state gel electrophoresis when examined at 4 degrees C. A testosterone binding protein with high binding affinity (K (a) = 0.98 +/- 0.5 x 10(8) M(-1)) and low to moderate binding capacity (B (max) = 7.58 +/- 4.2 x 10(-5) M) was observed in male hatchlings. An oestradiol binding protein with high affinity (K (a) = 0.35 +/- 1.8 x 10(8) M(-1)) and low to moderate binding capacity (B (max) = 0.16 +/- 0.5 x 10(-4) M) was identified in female hatchlings. This study confirmed that sex steroid binding proteins (SSBPs) become inactivate in both sexes at 36 degrees C, the maximum body temperature of sea turtle hatchlings at emergence. The inactivation of SSBPs at this temperature indicates that sex steroid hormones circulate freely in the body of the green turtles and are biologically available in the blood plasma. This observation is consistent with female and male hatchling C. mydas having different physiological (hormonal) and developmental requirements around the time of emergence. Moreover, concurrently conducted competition studies showed that sex steroids including testosterone and oestradiol do compete for binding sites in both male and female C. mydas hatchling plasma. Competition also occurred between testosterone and dihydrotestosterone for binding sites in the male C. mydas plasma. However, competition studies in the plasma of female hatchling C. mydas demonstrate that oestrone does not compete with oestradiol for binding sites.

  7. Effect of heparin administration on plasma binding of benzodiazepines.

    OpenAIRE

    1980-01-01

    1 The effect of intravenous administration of 100 units of heparin on plasma of diazepam, chlordiazepoxide, oxazepam and lorazepam was examined in fourteen normal subjects and five patients with cirrhosis. 2 In normal non-fasted subjects heparin caused a rapid 150--250% rise in the free fraction of diazepam, chlordiazepoxide and oxazepam but no change of lorazepam. The changes in free fraction were slightly smaller, but still significant, when the subjects were fasted. 3 These change in free ...

  8. Prostasomes of canine seminal plasma - zinc-binding ability and effects on motility characteristics and plasma membrane integrity of spermatozoa.

    Science.gov (United States)

    Mogielnicka-Brzozowska, M; Strzeżek, R; Wasilewska, K; Kordan, W

    2015-06-01

    Prostasomes are small lipid membrane-confined vesicles that are involved in various fertilization-related processes. The aim of this study was to demonstrate canine seminal plasma prostasomes' ability to bind zinc ions, as well as examining their effects on sperm motility characteristics and plasma membrane integrity during cold storage. Ejaculates, collected from five cross-bred dogs (n = 50), were subjected to ultracentrifugation followed by gel filtration (GF) on a Superose 6 column. Prostasomes appeared as a single fraction in the elution profile. Transmission electron microscopy (TEM) analysis of canine prostasomes revealed the presence of membrane vesicles with diameters ranging from 20.3 to 301 nm. The zinc-affinity chromatography on a Chelating Sepharose Fast Flow - Zn(2 +) showed that from 93 to 100% of the prostasome proteins bind zinc ions (P(+) Zn). SDS-PAGE revealed that canine P(+) Zn comprised four protein bands, with low molecular weights (10.2-12 kDa). We have also shown a positive effect of prostasomes (p spermatozoa with intact plasma membrane (SYBR/PI dual staining) and intact acrosome (Giemsa stained), after 2 h storage at 5°C, was showed, in variant A (1.5% of total seminal plasma protein) and B, when compared with Control and variant C (2.5% of total seminal plasma protein). The prostasomes' effect on motility and plasma membrane integrity of canine cold-stored spermatozoa may be related to their ability to bind zinc ions and regulate their availability to the sperm.

  9. Plasma levels of osteocalcin and retinol binding protein-4 in patients with medullary thyroid carcinoma

    Directory of Open Access Journals (Sweden)

    Jabar Lotfi

    2014-04-01

    Conclusion: According to difference between plasma levels of osteocalcin and retinol binding protein-4 in patients suffered of medullary thyroid carcinoma comparison with normal subjects, it can be said that, probably medullary thyroid carcinoma has effect on bone and adipose tissue metabolism, so osteocalcin and retinol binding protein-4 hormones have potential to be used for confirmation of diagnosis or following treatment of medullary thyroid carcinoma.

  10. Thyrotropin binding to porcine thyroid plasma membranes: kinetic and thermodynamic analyses.

    Science.gov (United States)

    Saltiel, A R; Thomas, C G; Nayfeh, S N

    1982-01-01

    Evaluation of TSH binding to plasma membranes of porcine thyroid revealed unique sensitivity to pH and temperature. Analysis of apparent equilibrium binding yielded a linear Scatchard plot at the optimal pH of 6.0, indicating one class of binding sites. At physiological pH 7.4 a curvilinear Scatchard plot was obtained, resolved by computer analysis into two classes of binding sites of different affinities and capacities. Treatment of membranes with phospholipase C resulted in a 20% decrease in the number of high affinity sites, but no change occurred in binding affinity. In contrast, low affinity sites were not altered. To evaluate the significance of the curvilinear Scatchard plot, the kinetics of association were examined. The intrinsic Kd (kd/ka) was 0.20 nM, a value essentially equivalent to that of the high affinity binding component. The 'negative cooperativity' model of hormone binding was evaluated by examining the effect of excess unlabeled TSH on dissociation rate. Dissociation of bound 125I-labeled TSH was biphasic, and was enhanced by unlabeled hormone, regardless of whether the membranes were prelabeled at pH 6.0 or 7.4. This effect was not correlated with curvilinear Scatchard plots, and therefore not proof of negative cooperativity. Binding sites for TSH were further distinguished by their sensitivity to temperature. A van't Hoff plot of temperature dependence of the apparent Kd of the high affinity site was linear from 4 to 37 degrees C. In contrast, the apparent Kd of low affinity binding did not vary with respect to temperature. These results demonstrate that there are at least two independent binding sites for TSH on porcine thyroid plasma membranes, distinguishable by their equilibrium binding properties.

  11. Haptoglobin-related protein is a high-affinity hemoglobin-binding plasma protein

    DEFF Research Database (Denmark)

    Nielsen, Marianne Jensby; Petersen, Steen Vang; Jacobsen, Christian

    2006-01-01

    Haptoglobin-related protein (Hpr) is a primate-specific plasma protein associated with apolipoprotein L-I (apoL-I)-containing high-density lipoprotein (HDL) particles shown to be a part of the innate immune defense. Despite the assumption hitherto that Hpr does not bind to hemoglobin, the present...

  12. Binding of von Willebrand factor and plasma proteins to the eggshell of Schistosoma mansoni

    NARCIS (Netherlands)

    Dewalick, Saskia; Hensbergen, Paul J; Bexkens, Michiel L; Grosserichter-Wagener, Christina; Hokke, Cornelis H; Deelder, André M; de Groot, Philip G; Tielens, Aloysius G M; van Hellemond, Jaap J

    2014-01-01

    Schistosoma mansoni eggs have to cross the endothelium and intestinal wall to leave the host and continue the life cycle. Mechanisms involved in this essential step are largely unknown. Here we describe direct binding to the S. mansoni eggshell of von Willebrand factor and other plasma proteins invo

  13. Binding of von Willebrand factor and plasma proteins to the eggshell of Schistosoma mansoni

    NARCIS (Netherlands)

    Dewalick, Saskia; Hensbergen, Paul J; Bexkens, Michiel L; Grosserichter-Wagener, Christina; Hokke, Cornelis H; Deelder, André M; de Groot, Philip G; Tielens, Aloysius G M; van Hellemond, Jaap J

    Schistosoma mansoni eggs have to cross the endothelium and intestinal wall to leave the host and continue the life cycle. Mechanisms involved in this essential step are largely unknown. Here we describe direct binding to the S. mansoni eggshell of von Willebrand factor and other plasma proteins

  14. MCP-1 binds to oxidized LDL and is carried by lipoprotein(a) in human plasma

    Science.gov (United States)

    Wiesner, Philipp; Tafelmeier, Maria; Chittka, Dominik; Choi, Soo-Ho; Zhang, Li; Byun, Young Sup; Almazan, Felicidad; Yang, Xiaohong; Iqbal, Navaid; Chowdhury, Punam; Maisel, Alan; Witztum, Joseph L.; Handel, Tracy M.; Tsimikas, Sotirios; Miller, Yury I.

    2013-01-01

    Lipoprotein oxidation plays an important role in pathogenesis of atherosclerosis. Oxidized low density lipoprotein (OxLDL) induces profound inflammatory responses in vascular cells, such as production of monocyte chemoattractant protein-1 (MCP-1) [chemokine (C-C motif) ligand 2], a key chemokine in the initiation and progression of vascular inflammation. Here we demonstrate that OxLDL also binds MCP-1 and that the OxLDL-bound MCP-1 retains its ability to recruit monocytes. A human MCP-1 mutant in which basic amino acids Arg-18 and Lys-19 were replaced with Ala did not bind to OxLDL. The MCP-1 binding to OxLDL was inhibited by the monoclonal antibody E06, which binds oxidized phospholipids (OxPLs) in OxLDL. Because OxPLs are carried by lipoprotein(a) [Lp(a)] in human plasma, we tested to determine whether Lp(a) binds MCP-1. Recombinant wild-type but not mutant MCP-1 added to human plasma bound to Lp(a), and its binding was inhibited by E06. Lp(a) captured from human plasma contained MCP-1 and the Lp(a)-associated endogenous MCP-1 induced monocyte migration. These results demonstrate that OxLDL and Lp(a) bind MCP-1 in vitro and in vivo and that OxPLs are major determinants of the MCP-1 binding. The association of MCP-1 with OxLDL and Lp(a) may play a role in modulating monocyte trafficking during atherogenesis. PMID:23667177

  15. Binding and Fusion of Extracellular Vesicles to the Plasma Membrane of Their Cell Targets.

    Science.gov (United States)

    Prada, Ilaria; Meldolesi, Jacopo

    2016-08-09

    Exosomes and ectosomes, extracellular vesicles of two types generated by all cells at multivesicular bodies and the plasma membrane, respectively, play critical roles in physiology and pathology. A key mechanism of their function, analogous for both types of vesicles, is the fusion of their membrane to the plasma membrane of specific target cells, followed by discharge to the cytoplasm of their luminal cargo containing proteins, RNAs, and DNA. Here we summarize the present knowledge about the interactions, binding and fusions of vesicles with the cell plasma membrane. The sequence initiates with dynamic interactions, during which vesicles roll over the plasma membrane, followed by the binding of specific membrane proteins to their cell receptors. Membrane binding is then converted rapidly into fusion by mechanisms analogous to those of retroviruses. Specifically, proteins of the extracellular vesicle membranes are structurally rearranged, and their hydrophobic sequences insert into the target cell plasma membrane which undergoes lipid reorganization, protein restructuring and membrane dimpling. Single fusions are not the only process of vesicle/cell interactions. Upon intracellular reassembly of their luminal cargoes, vesicles can be regenerated, released and fused horizontally to other target cells. Fusions of extracellular vesicles are relevant also for specific therapy processes, now intensely investigated.

  16. In vitro plasma protein binding and aqueous aggregation behavior of astaxanthin dilysinate tetrahydrochloride.

    Science.gov (United States)

    Zsila, Ferenc; Fitos, Ilona; Bikádi, Zsolt; Simonyi, Miklós; Jackson, Henry L; Lockwood, Samuel F

    2004-11-01

    The tetrahydrochloride salt of astaxanthin di-L-lysinate (lys(2)AST) is a highly water-dispersible astaxanthin-amino acid conjugate, with an aqueous dispersibility of > or = 181.6 mg/mL. The statistical mixture of stereoisomers has been well characterized as an aqueous-phase superoxide anion scavenger, effective at micromolar (microM) concentrations. In the current study, the aqueous aggregation behavior and in vitro plasma protein binding [with fatty-acid-free human serum albumin (HSA) and alpha(1)-acid glycoprotein (AGP)] were investigated with a suite of techniques, including circular dichroism (CD) and UV-vis spectroscopy, ultrafiltration, competitive ligand displacement, and fluorescence quenching. Induced CD bands obtained in Ringer buffer solution of HSA demonstrated high affinity monomeric binding of the compound at low ligand per protein (L/P) ratios (in aqueous solution alone the carotenoid molecules formed card-pack aggregates). The binding constant ( approximately 10(6)M(-1)) and the binding stoichiometry (approximately 0.2 per albumin molecule) were calculated from CD titration data. CD displacement and ultrafiltration experiments performed with marker ligands of HSA indicated that the ligand binding occurred at a site distinct from the main drug binding sites of HSA (i.e., Sites I and II). At intermediate L/P ratios, both monomeric and aggregated ("chirally complexed") binding occurred simultaneously at distinct sites of the protein. At high L/P ratios, chiral complexation predominantly occurred on the asymmetric protein template. The tentative location of the chirally-complexed aggregation on the HSA template was identified as the large interdomain cleft of HSA, where carotenoid derivatives have been found to bind previously. Only weak binding to AGP was observed. These results suggest that parenteral use of this highly potent, water-dispersible astaxanthin-amino acid conjugate will result in plasma protein association, and plasma protein binding at

  17. Autolytic Activity and Plasma Binding Study of Aap, a Novel Minor Autolysin of Streptococcus pneumoniae

    Directory of Open Access Journals (Sweden)

    Ramina Mahboobi

    2016-04-01

    Full Text Available Pneumococcal autolysins are enzymes involved in cell wall turnover and cellular division physiologically. They have been found to be involved in the pneumococcus pathogenesis. The aim of this study was to identify the autolytic activity of Spr1754 as a novel protein of Streptococcus pneumoniae. Moreover, the binding of the recombinant protein to plasma proteins was also determined. The spr1754 gene was amplified by PCR and cloned into the pET21a(+ prokaryotic expression vector. The constructed pET21a(+/spr1754 recombinant plasmid was transformed into E. coli Origami (DE3 and induced using IPTG. The recombinant protein of Spr1754 was purified by Ni-NTA affinity chromatography and confirmed by SDS-PAGE and Western blot analysis using anti-His tag monoclonal antibody. Autolytic activity and the ability of the recombinant protein in binding to plasma proteins were performed using zymogram analysis and western blot, respectively. The spr1754 with expected size was cloned and overexpressed in Escherichia coli Origami (DE3, successfully. After purification of the Spr1754 recombinant protein, the autolytic activity was observed by zymography. Of the four plasma proteins used in this study, binding of lactoferrin to Spr1754 recombinant protein was shown. The Spr1754 recombinant protein has a bifunctional activity, i.e., as being autolysin and lactoferrin binding and designated as Aap (autolytic/ adhesion/ pneumococcus. Nevertheless, characterization of the Aap needs to be followed using gene inactivation and cell wall localization.

  18. IgG Conformer's Binding to Amyloidogenic Aggregates.

    Directory of Open Access Journals (Sweden)

    Monichan Phay

    Full Text Available Amyloid-reactive IgGs isolated from pooled blood of normal individuals (pAbs have demonstrated clinical utility for amyloid diseases by in vivo targeting and clearing amyloidogenic proteins and peptides. We now report the following three novel findings on pAb conformer's binding to amyloidogenic aggregates: 1 pAb aggregates have greater activity than monomers (HMW species > dimers > monomers, 2 pAbs interactions with amyloidogenic aggregates at least partially involves unconventional (non-CDR interactions of F(ab regions, and 3 pAb's activity can be easily modulated by trace aggregates generated during sample processing. Specifically, we show that HMW aggregates and dimeric pAbs present in commercial preparations of pAbs, intravenous immunoglobulin (IVIg, had up to ~200- and ~7-fold stronger binding to aggregates of Aβ and transthyretin (TTR than the monomeric antibody. Notably, HMW aggregates were primarily responsible for the enhanced anti-amyloid activities of Aβ- and Cibacron blue-isolated IVIg IgGs. Human pAb conformer's binding to amyloidogenic aggregates was retained in normal human sera, and mimicked by murine pAbs isolated from normal pooled plasmas. An unconventional (non-CDR component to pAb's activity was indicated from control human mAbs, generated against non-amyloid targets, binding to aggregated Aβ and TTR. Similar to pAbs, HMW and dimeric mAb conformers bound stronger than their monomeric forms to amyloidogenic aggregates. However, mAbs had lower maximum binding signals, indicating that pAbs were required to saturate a diverse collection of binding sites. Taken together, our findings strongly support further investigations on the physiological function and clinical utility of the inherent anti-amyloid activities of monomeric but not aggregated IgGs.

  19. IgG Conformer's Binding to Amyloidogenic Aggregates

    Science.gov (United States)

    Phay, Monichan; Welzel, Alfred T.; Williams, Angela D.; McWilliams-Koeppen, Helen P.; Blinder, Veronika; O'Malley, Tiernan T.; Solomon, Alan; Walsh, Dominic M.; O'Nuallain, Brian

    2015-01-01

    Amyloid-reactive IgGs isolated from pooled blood of normal individuals (pAbs) have demonstrated clinical utility for amyloid diseases by in vivo targeting and clearing amyloidogenic proteins and peptides. We now report the following three novel findings on pAb conformer's binding to amyloidogenic aggregates: 1) pAb aggregates have greater activity than monomers (HMW species > dimers > monomers), 2) pAbs interactions with amyloidogenic aggregates at least partially involves unconventional (non-CDR) interactions of F(ab) regions, and 3) pAb's activity can be easily modulated by trace aggregates generated during sample processing. Specifically, we show that HMW aggregates and dimeric pAbs present in commercial preparations of pAbs, intravenous immunoglobulin (IVIg), had up to ~200- and ~7-fold stronger binding to aggregates of Aβ and transthyretin (TTR) than the monomeric antibody. Notably, HMW aggregates were primarily responsible for the enhanced anti-amyloid activities of Aβ- and Cibacron blue-isolated IVIg IgGs. Human pAb conformer's binding to amyloidogenic aggregates was retained in normal human sera, and mimicked by murine pAbs isolated from normal pooled plasmas. An unconventional (non-CDR) component to pAb's activity was indicated from control human mAbs, generated against non-amyloid targets, binding to aggregated Aβ and TTR. Similar to pAbs, HMW and dimeric mAb conformers bound stronger than their monomeric forms to amyloidogenic aggregates. However, mAbs had lower maximum binding signals, indicating that pAbs were required to saturate a diverse collection of binding sites. Taken together, our findings strongly support further investigations on the physiological function and clinical utility of the inherent anti-amyloid activities of monomeric but not aggregated IgGs. PMID:26367058

  20. Investigation of the selectivity of thrombin-binding aptamers for thrombin titration in murine plasma.

    Science.gov (United States)

    Trapaidze, Ana; Hérault, Jean-Pascal; Herbert, Jean-Marc; Bancaud, Aurélien; Gué, Anne-Marie

    2016-04-15

    Detection of thrombin in plasma raises timely challenges to enable therapeutic management of thrombosis in patients under vital threat. Thrombin binding aptamers represent promising candidates as sensing elements for the development of real-time thrombin biosensors; however implementation of such biosensor requires the clear understanding of thrombin-aptamer interaction properties in real-like environment. In this study, we used Surface Plasmon Resonance technique to answer the questions of specificity and sensitivity of thrombin detection by the thrombin-binding aptamers HD1, NU172 and HD22. We systematically characterized their properties in the presence of thrombin, as well as interfering molecular species such as the thrombin precursor prothrombin, thrombin in complex with some of its natural inhibitors, nonspecific serum proteins, and diluted plasma. Kinetic experiments show the multiple binding modes of HD1 and NU172, which both interact with multiple sites of thrombin with low nanomolar affinities and show little specificity of interaction for prothrombin vs. thrombin. HD22, on the other hand, binds specifically to thrombin exosite II and has no affinity to prothrombin at all. While thrombin in complex with some of its inhibitors could not be recognized by any aptamer, the binding of HD1 and NU172 properties is compromised by thrombin inhibitors alone, as well as with serum albumin. Finally, the complex nature of plasma was overwhelming for HD1, but we define conditions for the thrombin detection at 10nM range in 100-fold diluted plasma by HD22. Consequently HD22 showed key advantage over HD1 and NU172, and appears as the only alternative to design an aptasensor.

  1. Aluminium fluoride and magnesium, activators of heterotrimeric GTP-binding proteins, affect high-affinity binding of the fungal toxin fusicoccin to the fusicoccin-binding protein in oat root plasma membranes.

    NARCIS (Netherlands)

    de Boer, A.H.; Van der Molen, G.W.; Prins, H.B.A.; Korthout, H.A.A.J.; van der Hoeven, P.C.J.

    1994-01-01

    The fusicoccin-binding protein was solubilised from purified oat root plasma membranes. The solubilised protein retained full binding activity, provided that protease inhibitors were included. Sodium fluoride reduced the high-affinity [H-3]fusicoccin binding to almost zero in a concentration-depende

  2. Low serum levels of prohepcidin, but not hepcidin-25, are related to anemia in familial amyloidosis TTR V30M.

    NARCIS (Netherlands)

    Beirao, I.; Almeida, S.; Swinkels, D.W.; Costa, P.M.; Moreira, L.; Fonseca, I.; Freitas, C.; Cabrita, A.; Porto, G.

    2008-01-01

    Familial amyloidosis TTR V30M (FAP-I) usually presents as a sensorimotor and autonomic neuropathy. Anemia was first described in this disease more than 20 years ago and classified as an anemia of chronic disease. However, so far no studies have addressed the role of inflammatory proteins in this dis

  3. Proteomic Characterization of Zinc-Binding Proteins of Canine Seminal Plasma.

    Science.gov (United States)

    Mogielnicka-Brzozowska, M; Kowalska, N; Fraser, L; Kordan, W

    2015-12-01

    The zinc-binding proteins (ZnBPs) of the seminal plasma are implicated in different processes related to sperm-egg fusion. The aim of this study was to characterize the ZnBPs of canine seminal plasma using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and mass spectrometry. The ZnBPs were isolated from the ejaculates of five dogs by affinity chromatography and subjected to 2D-PAGE analysis. The acquired spots, detected across the gels, were analysed by mass spectrometry. Using 2D-PAGE analysis, it was shown that canine seminal plasma comprised about 46-57 zinc-binding polypeptides, with molecular mass ranging from 9.3 to 138.7 kDa and pI at pH 5.2-10.0. It was found that zinc-binding polypeptides of low molecular masses (9.3-19.0 kDa and pI at pH 6.1-10.0) were predominant in the seminal plasma, and seven polypeptides, with molecular masses ranging from 11.7 to 15.4 kDa and pI at pH 6.8-8.7, were characterized by high optical density values. In addition, analysis with mass spectrometry (LC-MS-MS/MS) revealed that the identified seven polypeptides are canine prostate-specific esterase (CPSE), which is the main proteolytic enzyme of the seminal plasma. The findings of this study indicate an important regulatory role of seminal plasma zinc ions in the functional activity of CPSE, which is of great significance for maintaining the normal function of canine prostate and the spermatozoa functions.

  4. Quartz crystal microbalance for the cardiac markers/antibodies binding kinetic measurements in the plasma samples

    Science.gov (United States)

    Agafonova, L. E.; Shumyantseva, V. V.; Archakov, A. I.

    2014-06-01

    The quartz crystal microbalance (QCM) was exploited for cardiac markers detection and kinetic studies of immunochemical reaction of cardiac troponin I (cTnI) and human heart fatty acid binding protein (H-FABP) with the corresponding monoclonal antibodies in undiluted plasma (serum) and standard solutions. The QCM technique allowed to dynamically monitor the kinetic differences in specific interactions and nonspecific sorption, without multiple labeling procedures and separation steps. The affinity binding process was characterized by the association (ka) and the dissociation (kd) kinetic constants and the equilibrium association (K) constant, all of which were obtained from experimental data.

  5. Why mammals more susceptible to the hepatotoxic microcystins than fish: evidences from plasma and albumin protein binding through equilibrium dialysis.

    Science.gov (United States)

    Zhang, Wei; Liang, Gaodao; Wu, Laiyan; Tuo, Xun; Wang, Wenjing; Chen, Jun; Xie, Ping

    2013-08-01

    To elucidate the interspecies variation of susceptibility to microcystins (MCs), fresh plasma and purified albumin from six kinds of mammals and fish were used in toxins-substances binding test. Protein contents in the test plasma were analyzed and the binding characteristics to MCs were compared. Two kinds of widely observed MCs, microcystin-LR (MC-LR) and microcystin-RR (MC-RR) were tested and data were collected through the method of equilibrium dialysis. It was found that total plasma protein and albumin content in mammals were nearly two times and four times higher than that in fish, respectively. In the test range of 0-100 μg/mL, binding rates of fish plasma to MCs were considered significant lower (p mammals. And human plasma demonstrated the highest binding rate in mammals. In all the test species, plasma protein binding rates of MC-RR were significantly higher than MC-LR (p 0.05). From the view of protein binding, it is concluded that both the variation of plasma protein composition and albumin binding characteristic could influence the existing form of MCs in circulation, change MCs utilization, alter MCs half-life and further contribute to the difference of susceptibility between mammals and fish.

  6. Ophthalmic manifestations in a Chinese family with familial amyloid polyneuropathy due to a TTR Gly83Arg mutation.

    Science.gov (United States)

    Liu, T; Zhang, B; Jin, X; Wang, W; Lee, J; Li, J; Yuan, H; Cheng, X

    2014-01-01

    To describe the characteristic ophthalmic phenotypes of a large Chinese family with familial amyloid polyneuropathy due to a missense mutation in transthyretin (TTR) (c.307 C>G). Twenty-seven individuals (12 affected, 15 unaffected) from a five-generation Chinese family underwent general medical examination and comprehensive ophthalmic examination, including best correct visual acuity, intraocular pressure measurements, Schirmer test, slitlamp examination, fundoscopy, and ocular ultrasonography. Histological examination of vitreous biopsies using Congo red staining and immunohistochemistry was performed. Cardiovascular magnetic resonance (CMR), electrocardiogram, and echocardiogram were used to evaluate cardiac amyloidosis. Electromyography was used to evaluate nerve function. All four exons of TTR were amplified by PCR, sequenced using a Bigdye terminator v3.1 cycle sequencing kit and analyzed on an ABI 3700XL Genetic Analyzer. All 12 affected individuals in the family had ocular manifestations, including severe vitreous opacities, secondary glaucoma, xerophthalmia, dyscoria, and attenuated retinal arteries. Congo red staining demonstrated amyloid deposits in the vitreous, and immunohistochemical staining confirmed the deposition of TTR proteins in the vitreous. Twelve individuals had polyneuropathy, and electromyography detected functional damage in peripheral nerves. One individual was diagnosed with cardiac amyloidosis by CMR. Direct sequencing revealed the heterozygous missense mutation in TTR (c.307 C>G p.Gly83Arg) in all 12 affected individuals. The mutation co-segregated with the disease phenotype and was absent in 100 normal controls. Vitreous opacity is very common in patients with the TTR Gly83Arg mutation; other clinical characteristics associated with the mutation include polyneuropathy and cardiac amyloidosis.

  7. IN VITRO IRREVERSIBLE BINDING AND DEGRADATION OF (R- AND (S-KETOPROFEN GLUCURONIDES TO PLASMA PROTEINS

    Directory of Open Access Journals (Sweden)

    PETER J. HAYBALL

    2007-01-01

    Full Text Available This study describes the in vitro degradation studies of the diastereomeric ketoprofen glucuronides, under physiological conditions (pH 7.4, 37°C, (R-ketoprofen glucuronide t½ = 30 min, (S-ketoprofen glucuronide t½ = 70 min and the irreversible binding of diastereomeric ketoprofen glucuronides (15 μg/ml to human serum albumin (HSA (289 μM and human plasma under physiological conditions (pH 7.4, 37ºC. The (R-ketoprofen glucuronide irreversibly bound to a greater extent in both human plasma and human serum albumin. This is the reverse to that found in previous studies. These findings further support the hypothesis that faster degradation of 1-O-acyl glucuronide (in this case the (R-diastereomer is associated with a greater extent of irreversible binding.

  8. Isolation and characterization of the plasma hyaluronan-binding protein (PHBP) gene (HABP2).

    Science.gov (United States)

    Sumiya, J; Asakawa, S; Tobe, T; Hashimoto, K; Saguchi, K; Choi-Miura, N H; Shimizu, Y; Minoshima, S; Shimizu, N; Tomita, M

    1997-11-01

    PHBP is a novel human plasma hyaluronan-binding protein that shows significant homology in amino acid sequence to hepatocyte growth factor activator. Two overlapping clones that encode the human plasma hyaluronan-binding protein (PHBP) gene (HABP2) were isolated and characterized. The PHBP gene spans 35 kb and is composed of 13 exons from 37 to 1,394 bp in size with consensus splice sites. The gene's regulatory sequences contain putative promoter elements, but no typical TATA box. Some exons of this gene showed significant similarities to those of coagulation factor XII, tissue-type plasminogen activator, and urokinase genes in nucleotide length and in intron phasing. We also report the chromosome mapping of this gene by fluorescence in situ hybridization (FISH) using a genomic DNA fragment as a probe. The PHBP gene (HABP2) was located on chromosome 10q25-q26.

  9. Extracellular Matrix Markers for Disease Progression and Follow-Up of Therapies in Familial Amyloid Polyneuropathy V30M TTR-Related

    Directory of Open Access Journals (Sweden)

    I. Cardoso

    2008-01-01

    Full Text Available Familial Amyloidotic Polyneuropathy (FAP is a disorder characterized by the extracellular deposition of fibrillar Transthyretin (TTR amyloid, with a special involvement of the peripheral nerve. Several extracellular matrix proteins have been found elevated in tissues from FAP patients, namely metalloproteinase-9 (MMP-9, neutrophil gelatinase associated lipocalin (NGAL and biglycan. In this work we assessed the levels of MMP-9, tissue inhibitor of metalloproteinase 1 (TIMP-1, NGAL, biglycan and chondroitin sulphate (CSPG in an FAP V30M TTR-related transgenic mouse model at different stages of TTR deposition and after two different treatment approaches to remove fibrillar deposits. Immunohistochemistry or RT-PCR analysis showed that biglycan was already increased in animals presenting TTR deposited in a non-fibrillar state, whereas MMP-9, TIMP-1, NGAL and CSPG were elevated only in mice with TTR amyloid deposits. Mice treated with doxycycline, a TTR fibril disrupter, presented lower levels of MMP-9, TIMP-1 and NGAL, suggestive of matrix recovery. Mice immunized with TTR Y78F to remove TTR deposition showed significantly lower levels of all the five tested markers, suggesting removal of fibrillar and non-fibrillar deposits. Cellular studies using oligomeric TTR showed induction of MMP-9 when compared to soluble TTR, large aggregates or fibrils. Furthermore, this induction was neutralized by an anti-receptor for advanced glycation end products (RAGE antibody, indicating RAGE engagement in this process. Further studies in a larger number of tissue samples will indicate the application of these ECM markers in parallel with Congo Red staining in tissue characterization of pre-clinical and clinical stages in FAP and other amyloidoses.

  10. Comparative binding mechanism of lupeol compounds with plasma proteins and its pharmacological importance.

    Science.gov (United States)

    Kallubai, Monika; Rachamallu, Aparna; Yeggoni, Daniel Pushparaju; Subramanyam, Rajagopal

    2015-04-01

    Lupeol, a triterpene, possesses beneficial effects like anti-inflammatory and anti-cancer properties. Binding of lupeol and its derivative (phytochemicals) to plasma proteins such as human serum albumin (HSA) and α-1-acid glycoprotein (AGP) is a major determinant in the disposition of drugs. Cytotoxic studies with mouse macrophages (RAW 246.7) and HeLa cell lines revealed anti-inflammatory and anti-cancer properties for both lupeol and lupeol derivative. Both molecules reduced the expression of pro-inflammatory cytokines in LPS induced macrophages. Further, apoptosis was observed in HeLa cell lines when they were incubated with these molecules for 24 h. The fluorescence quenching of HSA was observed upon titration with different concentrations of lupeol and lupeol derivative; their binding constants were found to be 3 ± 0.01 × 10(4) M(-1) and 6.2 ± 0.02 × 10(4) M(-1), with binding free energies of -6.59 kcal M(-1) and -7.2 kcal M(-1). With AGP, however, the lupeol and lupeol derivative showed binding constants of 0.9 ± 0.02 × 10(3) M(-1) and 2.7 ± 0.01 × 10(3) M(-1), with free energies of -4.6 kcal M(-1) and -5.1 kcal M(-1) respectively. Molecular displacement studies based on competition with site I-binding phenylbutazone (which binds site I of HSA) and ibuprofen (which binds site II) suggest that lupeol binds site II and the lupeol derivative site I. Molecular docking studies also confirmed that lupeol binds to the IIIA and the lupeol derivative to the IIA domain of HSA. Secondary structure changes were observed upon formation of HSA-lupeol/lupeol derivative complexes by circular dichroism spectroscopy. Molecular dynamics simulations support greater stability of HSA-lupeol and HSA-lupeol derivative complexes compared to that of HSA alone.

  11. Diclofenac plasma protein binding: PK-PD modelling in cardiac patients submitted to cardiopulmonary bypass

    Directory of Open Access Journals (Sweden)

    Auler Jr. J.O.

    1997-01-01

    Full Text Available Twenty-four surgical patients of both sexes without cardiac, hepatic, renal or endocrine dysfunctions were divided into two groups: 10 cardiac surgical patients submitted to myocardial revascularization and cardiopulmonary bypass (CPB, 3 females and 7 males aged 65 ± 11 years, 74 ± 16 kg body weight, 166 ± 9 cm height and 1.80 ± 0.21 m2 body surface area (BSA, and control, 14 surgical patients not submitted to CPB, 11 female and 3 males aged 41 ± 14 years, 66 ± 14 kg body weight, 159 ± 9 cm height and 1.65 ± 0.16 m2 BSA (mean ± SD. Sodium diclofenac (1 mg/kg, im Voltaren 75® twice a day was administered to patients in the Recovery Unit 48 h after surgery. Venous blood samples were collected during a period of 0-12 h and analgesia was measured by the visual analogue scale (VAS during the same period. Plasma diclofenac levels were measured by high performance liquid chromatography. A two-compartment open model was applied to obtain the plasma decay curve and to estimate kinetic parameters. Plasma diclofenac protein binding decreased whereas free plasma diclofenac levels were increased five-fold in CPB patients. Data obtained for analgesia reported as the maximum effect (EMAX were: 25% VAS (CPB vs 10% VAS (control, P<0.05, median measured by the visual analogue scale where 100% is equivalent to the highest level of pain. To correlate the effect versus plasma diclofenac levels, the EMAX sigmoid model was applied. A prolongation of the mean residence time for maximum effect (MRTEMAX was observed without any change in lag-time in CPB in spite of the reduced analgesia reported for these patients, during the time-dose interval. In conclusion, the extent of plasma diclofenac protein binding was influenced by CPB with clinically relevant kinetic-dynamic consequences

  12. Assessment of coronary reperfusion in patients with myocardial infarction using fatty acid binding protein concentrations in plasma

    NARCIS (Netherlands)

    M.J.M. de Groot; A.M.M. Muijtjens; M.L. Simoons (Maarten); W.T. Hermens (Wim); J.F.C. Glatz

    2001-01-01

    textabstractOBJECTIVE: To examine whether successful coronary reperfusion after thrombolytic treatment in patients with confirmed acute myocardial infarction can be diagnosed from the plasma marker fatty acid binding protein (FABP), for either acute clinical decision making or retrospective purposes

  13. Curcumin could reduce the monomer of TTR with Tyr114Cys mutation via autophagy in cell model of familial amyloid polyneuropathy

    Directory of Open Access Journals (Sweden)

    Li H

    2014-10-01

    Full Text Available Hui Li,1,* Yu Zhang,1,* Li Cao,1 Ran Xiong,1 Bei Zhang,1 Li Wu,1 Zongbo Zhao,1 Sheng-Di Chen1,2 1Department of Neurology and Institute of Neurology, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, 2Key Laboratory of Stem Cell Biology and Laboratory of Neurodegenerative Diseases, Institute of Health Science, Shanghai Institutes of Biological Sciences, Chinese Academy of Science, and Shanghai Jiao Tong University School of Medicine, Shanghai, People’s Republic of China *These authors contributed equally to this work Abstract: Transthyretin (TTR familial amyloid polyneuropathy (FAP is an autosomal ­dominant inherited neurodegenerative disorder caused by various mutations in the transthyretin gene. We aimed to identify the mechanisms underlying TTR FAP with Tyr114Cys (Y114C mutation. Our study showed that TTR Y114C mutation led to an increase in monomeric TTR and impaired autophagy. Treatment with curcumin resulted in a significant decrease of monomeric TTR by recovering autophagy. Our research suggests that impairment of autophagy might be involved in the pathogenesis of TTR FAP with Y114C mutation, and curcumin might be a potential therapeutic approach for TTR FAP. Keywords: curcumin, familial amyloid polyneuropathy, transthyretin, autophagy

  14. Binding of plasma proteins to titanium dioxide nanotubes with different diameters.

    Science.gov (United States)

    Kulkarni, Mukta; Flašker, Ajda; Lokar, Maruša; Mrak-Poljšak, Katjuša; Mazare, Anca; Artenjak, Andrej; Čučnik, Saša; Kralj, Slavko; Velikonja, Aljaž; Schmuki, Patrik; Kralj-Iglič, Veronika; Sodin-Semrl, Snezna; Iglič, Aleš

    2015-01-01

    Titanium and titanium alloys are considered to be one of the most applicable materials in medical devices because of their suitable properties, most importantly high corrosion resistance and the specific combination of strength with biocompatibility. In order to improve the biocompatibility of titanium surfaces, the current report initially focuses on specifying the topography of titanium dioxide (TiO2) nanotubes (NTs) by electrochemical anodization. The zeta potential (ζ-potential) of NTs showed a negative value and confirmed the agreement between the measured and theoretically predicted dependence of ζ-potential on salt concentration, whereby the absolute value of ζ-potential diminished with increasing salt concentrations. We investigated binding of various plasma proteins with different sizes and charges using the bicinchoninic acid assay and immunofluorescence microscopy. Results showed effective and comparatively higher protein binding to NTs with 100 nm diameters (compared to 50 or 15 nm). We also showed a dose-dependent effect of serum amyloid A protein binding to NTs. These results and theoretical calculations of total available surface area for binding of proteins indicate that the largest surface area (also considering the NT lengths) is available for 100 nm NTs, with decreasing surface area for 50 and 15 nm NTs. These current investigations will have an impact on increasing the binding ability of biomedical devices in the body leading to increased durability of biomedical devices.

  15. Characterization of GTP binding and hydrolysis in plasma membranes of zucchini

    Science.gov (United States)

    Perdue, D. O.; Lomax, T. L.

    1992-01-01

    We have investigated the possibility that G-protein-like entities may be present in the plasma membrane (PM) of zucchini (Cucurbita pepo L.) hypocotyls by examining a number of criteria common to animal and yeast G-proteins. The GTP binding and hydrolysis characteristics of purified zucchini PM are similar to the characteristics of a number of known G-proteins. Our results demonstrate GTP binding to a single PM site having a Kd value between 16-31 nM. This binding has a high specificity for guanine nucleotides, and is stimulated by Mg2+, detergents, and fluoride or aluminium ions. The GTPase activity (Km = 0.49 micromole) of zucchini PM shows a sensitivity to NaF similar to that seen for other G-proteins. Localization of GTP mu 35S binding to nitrocellulose blots of proteins separated by SDS-PAGE indicates a 30-kDa protein as the predominant GTP-binding species in zucchini PM. Taken together, these data indicate that plant PM contains proteins which are biochemically similar to previously characterized G-proteins.

  16. SDS-binding assay based on tyrosine fluorescence as a tool to determine binding properties of human serum albumin in blood plasma

    Science.gov (United States)

    Zhdanova, Nadezda; Shirshin, Evgeny; Fadeev, Victor; Priezzhev, Alexander

    2016-04-01

    Among all plasma proteins human serum albumin (HSA) is the most studied one as it is the main transport protein and can bind a wide variety of ligands especially fatty acids (FAs). The concentration of FAs bound to HSA in human blood plasma differs by three times under abnormal conditions (fasting, physical exercises or in case of social important diseases). In the present study a surfactant sodium dodecyl sulfate (SDS) was used to simulate FAs binding to HSA. It was shown that the increase of Tyr fluorescence of human blood plasma due to SDS addition can be completely explained by HSA-SDS complex formation. Binding parameters of SDS-HSA complex (average number of sites and apparent constant of complex formation) were determined from titration curves based on tyrosine (Tyr) fluorescence.

  17. Piperine Decreases Binding of Drugs to Human Plasma and Increases Uptake by Brain Microvascular Endothelial Cells.

    Science.gov (United States)

    Dubey, Raghvendra K; Leeners, Brigitte; Imthurn, Bruno; Merki-Feld, Gabriele Susanne; Rosselli, Marinella

    2017-09-26

    We previously reported that piperine, an active alkaloidal principal of black and long peppers, enhances drug bioavailability by inhibiting drug metabolism. Another mechanism influencing drug availability/uptake is its free fraction. Since piperine is highly lipophilic, we hypothesize that it could also interact with drugs through binding displacement and influence their bioavailability. Accordingly, using equilibrium dialysis, we investigated whether piperine alters the binding of model drug ligands, that is flunitrazepam, diazepam, warfarin, salicylic acid, propranolol, lidocaine, and disopyramide to human plasma (n = 4). Since alterations in binding influence drug disposition, we also studied the effects of piperine on the uptake of plasma bound (3) H-propranolol and (14) C-warfarin by cultured bovine brain microvascular endothelial cells (BMECs). Piperine (1-1000 μM) increased the free fraction (fu) of both albumin and alpha-acid glycoprotein bound drugs in a concentration-dependent manner (p < 0.01). Moreover, piperine (10 μM) increased the uptake of (3) H-propranolol and (14) C-warfarin by BMECs (p < 0.01). In conclusion, our findings provide the first evidence that piperine displaces plasma bound drugs from both albumin and alpha-acid glycoprotein and facilitates drug uptake across biological membranes (e.g. BMEC). Moreover, it is feasible that piperine may similarly facilitate the transport of drugs into tissues, in vivo, and alter both pharmacokinetics and pharmacodynamics of administered drugs. Copyright © 2017 John Wiley & Sons, Ltd. Copyright © 2017 John Wiley & Sons, Ltd.

  18. Irreversible binding of an anticancer compound (BI-94) to plasma proteins.

    Science.gov (United States)

    Gautam, Nagsen; Thakare, Rhishikesh; Rana, Sandeep; Natarajan, Amarnath; Alnouti, Yazen

    2015-01-01

    1. We investigated the mechanisms responsible for the in vivo instability of a benzofurazan compound BI-94 (NSC228148) with potent anti-cancer activity. 2. BI-94 was stable in MeOH, water, and in various buffers at pHs 2.5-5, regardless of the buffer composition. In contrast, BI-94 was unstable in NaOH and at pHs 7-9, regardless of the buffer composition. BI-94 disappeared immediately after spiking into mice, rat, monkey, and human plasma. BI-94 stability in plasma can be only partially restored by acidifying it, which indicated other mechanisms in addition to pH for BI-94 instability in plasma. 3. BI-94 formed adducts with the trapping agents, glutathione (GSH) and N-acetylcysteine (NAC), in vivo and in vitro via nucleophilic aromatic substitution reaction. The kinetics of adduct formation showed that neutral or physiological pHs enhanced and accelerated GSH and NAC adduct formation with BI-94, whereas acidic pHs prevented it. Therefore, physiological pHs not only altered BI-94 chemical stability but also enhanced adduct formation with endogenous nucleophiles. In addition, adduct formation with human serum albumin-peptide 3 (HSA-T3) at the Cys34 position was demonstrated. 4. In conclusion, BI-94 was unstable at physiological conditions due to chemical instability and irreversible binding to plasma proteins.

  19. Reliability of plasma lipopolysaccharide-binding protein (LBP) from repeated measures in healthy adults.

    Science.gov (United States)

    Citronberg, Jessica S; Wilkens, Lynne R; Lim, Unhee; Hullar, Meredith A J; White, Emily; Newcomb, Polly A; Le Marchand, Loïc; Lampe, Johanna W

    2016-09-01

    Plasma lipopolysaccharide-binding protein (LBP), a measure of internal exposure to bacterial lipopolysaccharide, has been associated with several chronic conditions and may be a marker of chronic inflammation; however, no studies have examined the reliability of this biomarker in a healthy population. We examined the temporal reliability of LBP measured in archived samples from participants in two studies. In Study one, 60 healthy participants had blood drawn at two time points: baseline and follow-up (either three, six, or nine months). In Study two, 24 individuals had blood drawn three to four times over a seven-month period. We measured LBP in archived plasma by ELISA. Test-retest reliability was estimated by calculating the intraclass correlation coefficient (ICC). Plasma LBP concentrations showed moderate reliability in Study one (ICC 0.60, 95 % CI 0.43-0.75) and Study two (ICC 0.46, 95 % CI 0.26-0.69). Restricting the follow-up period improved reliability. In Study one, the reliability of LBP over a three-month period was 0.68 (95 % CI: 0.41-0.87). In Study two, the ICC of samples taken ≤seven days apart was 0.61 (95 % CI 0.29-0.86). Plasma LBP concentrations demonstrated moderate test-retest reliability in healthy individuals with reliability improving over a shorter follow-up period.

  20. Fluorescence Enhancement of Fluorescein Isothiocyanate-Labeled Protein A Caused by Affinity Binding with Immunoglobulin G in Bovine Plasma

    Directory of Open Access Journals (Sweden)

    Kiyotaka Sakai

    2009-10-01

    Full Text Available Fluorescence enhancement of fluorescein isothiocyanate-labeled protein A (FITC-protein A caused by the binding with immunoglobulin G (IgG in bovine plasma was studied. FITC-protein A was immobilized onto a glass surface by covalent bonds. An increase in fluorescence intensity was dependent on IgG concentration ranging from 20 to 78 μg/mL in both phosphate buffer saline and bovine plasma. This method requires no separation procedure, and the reaction time is less than 15 min. A fluorescence enhancement assay by the affinity binding of fluorescence-labeled reagent is thus available for the rapid determination of biomolecules in plasma.

  1. Plasma protein-binding parameters of prednisolone in immune disease patients receiving long-term prednisone therapy.

    Science.gov (United States)

    Wagner, J G; Wexler, D; Ağabeyoğlu, I T; Bergstrom, R F; Sakmar, E; Kay, D R

    1981-04-01

    Prednisone and prednisolone bind in plasma to albumin and transcortin. In am attempt to determine whether prednisone side effects and/or type of disease correlated with prednisolone plasma protein binding, multiple plasma samples from 17 patients (three asthma, eight SLE, three RA, two PSS, one PAN) receiving long-term prednisone therapy were monitored during an interval between two prednisone doses. Prednisolone plasma protein binding was nonlinear and exhibited large intrapatient and interpatient variability. For the group, mean association constants of the prednisolone-albumin complex and the prednisolone-transcortin complex were 2.3 X 10(3) M-1 and 2.9 X 10(7) M-1, with coefficients of variation of 82% and 127%, respectively. SLE patients tended to have lower mean prednisolone association constants for albumin and transcortin than did other patients. The presence of corticosteroid side effects did not correlate with prednisolone plasma protein-binding parameters. The wide range of prednisolone free fraction noted in plasma from patients who achieved comparable total prednisolone plasma concentrations implies that administration of a uniform prednisone dose will not lead to a predictable clinical response.

  2. Maternal Plasma Retinol Binding Protein 4 in Acute Pyelonephritis during Pregnancy

    Science.gov (United States)

    Vaisbuch, Edi; Romero, Roberto; Mazaki-Tovi, Shali; Kusanovic, Juan Pedro; Chaiworapongsa, Tinnakorn; Dong, Zhong; Kim, Sun Kwon; Ogge, Giovanna; Gervasi, Maria Teresa; Hassan, Sonia S.

    2010-01-01

    Objective Adipokines have been implicated in metabolic regulation and the immune response thus providing a molecular mechanism for the interaction between these two systems. Retinol binding protein 4 (RBP4) is a novel adipokine that plays a role in the pathophysiology of obesity-induced insulin resistance, as well as in the modulation of inflammation. The aim of this study was to determine whether there are changes in maternal plasma concentrations of RBP4 in pregnant women with acute pyelonephritis. Study design This cross-sectional study included pregnant women in the following groups: 1) normal pregnancy (n=80); 2) pyelonephritis (n=39). Maternal plasma RBP4 concentrations were determined by enzyme-linked immunoassays. Non-parametric statistics were used for analyses. Results 1) The median maternal plasma RBP4 concentration was lower in patients with acute pyelonephritis than in those with a normal pregnancy (3709.6 ng/mL, IQR 2917.7-5484.2 vs. 9167.6 ng/mL, IQR 7496.1-10384.1, ppyelonephritis who had a positive blood culture and those with a negative culture (3285.3 ng/mL, IQR 2274.1-4741.1 vs. 3922.6 ng/mL, IQR 3126.8-5547.1, respectively, p=0.2); and 3) lower maternal plasma RBP4 concentrations were independently associated with pyelonephritis after adjustment for confounding factors. Conclusions In contrast to what has been reported in preeclampsia, acute pyelonephritis during pregnancy is associated with lower maternal plasma RBP4 concentrations than in normal pregnancy. This finding suggests that the acute maternal inflammatory process associated with pyelonephritis is fundamentally different from that of the chronic systemic inflammatory process suggested in preeclampsia, in which RBP4 concentrations were found to be elevated. PMID:20163326

  3. New locus (ttr) in Escherichia coli K-12 affecting sensitivity to bacteriophage T2 and growth on oleate as the sole carbon source.

    Science.gov (United States)

    Morona, R; Henning, U

    1986-11-01

    The nature of resistance to phage T2 in Escherichia coli K-12 was investigated by analyzing a known phage T2-resistant mutant and by isolating new T2-resistant mutants. It was found that mutational alterations at two loci, ompF (encoding the outer membrane protein OmpF) and ttr (T-two resistance), are needed to give full resistance to phage T2. A ttr::Tn10 mutation was isolated and was mapped between aroC and dsdA, where the fadL gene (required for long-chain fatty acid transport) is located. The receptor affected by ttr was the major receptor used by phage T2 and was located in the outer membrane. Phage T2 was thus able to use two outer membrane proteins as receptors. All strains having a ttr::Tn10 allele and most of the independently isolated phage T2-resistant mutants were unable to grow on oleate as the sole carbon and energy source, i.e., they had the phenotype of fadL mutants. The gene fadL is known to encode an inner membrane protein. The most likely explanation is that fadL and ttr are in an operon and that ttr encodes an outer membrane protein which functions in translocating long-chain fatty acids across the outer membrane and also as a receptor for phage T2.

  4. Partitioning, diffusion, and ligand binding of raft lipid analogs in model and cellular plasma membranes.

    Science.gov (United States)

    Sezgin, Erdinc; Levental, Ilya; Grzybek, Michal; Schwarzmann, Günter; Mueller, Veronika; Honigmann, Alf; Belov, Vladimir N; Eggeling, Christian; Coskun, Unal; Simons, Kai; Schwille, Petra

    2012-07-01

    Several simplified membrane models featuring coexisting liquid disordered (Ld) and ordered (Lo) lipid phases have been developed to mimic the heterogeneous organization of cellular membranes, and thus, aid our understanding of the nature and functional role of ordered lipid-protein nanodomains, termed "rafts". In spite of their greatly reduced complexity, quantitative characterization of local lipid environments using model membranes is not trivial, and the parallels that can be drawn to cellular membranes are not always evident. Similarly, various fluorescently labeled lipid analogs have been used to study membrane organization and function in vitro, although the biological activity of these probes in relation to their native counterparts often remains uncharacterized. This is particularly true for raft-preferring lipids ("raft lipids", e.g. sphingolipids and sterols), whose domain preference is a strict function of their molecular architecture, and is thus susceptible to disruption by fluorescence labeling. Here, we analyze the phase partitioning of a multitude of fluorescent raft lipid analogs in synthetic Giant Unilamellar Vesicles (GUVs) and cell-derived Giant Plasma Membrane Vesicles (GPMVs). We observe complex partitioning behavior dependent on label size, polarity, charge and position, lipid headgroup, and membrane composition. Several of the raft lipid analogs partitioned into the ordered phase in GPMVs, in contrast to fully synthetic GUVs, in which most raft lipid analogs mis-partitioned to the disordered phase. This behavior correlates with the greatly enhanced order difference between coexisting phases in the synthetic system. In addition, not only partitioning, but also ligand binding of the lipids is perturbed upon labeling: while cholera toxin B binds unlabeled GM1 in the Lo phase, it binds fluorescently labeled GMI exclusively in the Ld phase. Fluorescence correlation spectroscopy (FCS) by stimulated emission depletion (STED) nanoscopy on intact

  5. Platelets and erythrocyte-bound platelets bind infectious HIV-1 in plasma of chronically infected patients.

    Science.gov (United States)

    Beck, Zoltan; Jagodzinski, Linda L; Eller, Michael A; Thelian, Doris; Matyas, Gary R; Kunz, Anjali N; Alving, Carl R

    2013-01-01

    Chronic HIV-1 infection is associated with persistent viremia in most patients, but it remains unclear how free virus may survive the potential hostile effects of plasma. We investigated whether sites might exist on the surfaces of circulating blood cells for protection of infectious HIV-1 particles. Red blood cells (RBC) either from blood of uninfected normal individuals, or from blood obtained without EDTA from chronically infected HIV-1 patients, invariably contained a small number of RBC having attached platelets as determined by flow cytometry, light microscopy, and immunofluorescence microscopy. After mixing normal RBC with platelet-rich plasma, discrete populations of RBC, platelets, and complexes of platelets attached to RBC were purified by fluorescence-activated cell sorting. Upon incubation of purified cells or platelets with HIV-1 followed by washing and co-incubation with CD4-positive peripheral blood mononuclear cells (PBMC), platelets, and platelet-RBC complexes, but not platelet-free RBC, caused infection of PBMC. Infection was prevented by pre-treating the platelet-RBC complexes with EDTA. Plasma and RBC (comprising a RBC/platelet-RBC mixture) from chronically infected patients with low viral loads were also co-incubated with PBMC ex vivo to determine the presence of infectious HIV-1. All freshly isolated plasmas from the HIV-1-infected donors, obtained in the absence of anticoagulant, were noninfectious. Interestingly, the RBC from most of the patients caused cell-cell infection of PBMC that was prevented by stripping the RBC with EDTA. A monoclonal antibody to DC-SIGN partially inhibited cell-cell HIV-1 infection of PBMC by normal RBC pre-incubated with platelets and HIV-1. We conclude: (a) platelet-free EDTA-free plasma from chronically infected HIV-1 patients, although containing viral RNA, is an environment that lacks detectable infectious HIV-1; (b) platelets and platelet-RBC complexes, but not purified RBC, bind infectious HIV-1; (c) DC

  6. Platelets and erythrocyte-bound platelets bind infectious HIV-1 in plasma of chronically infected patients.

    Directory of Open Access Journals (Sweden)

    Zoltan Beck

    Full Text Available Chronic HIV-1 infection is associated with persistent viremia in most patients, but it remains unclear how free virus may survive the potential hostile effects of plasma. We investigated whether sites might exist on the surfaces of circulating blood cells for protection of infectious HIV-1 particles. Red blood cells (RBC either from blood of uninfected normal individuals, or from blood obtained without EDTA from chronically infected HIV-1 patients, invariably contained a small number of RBC having attached platelets as determined by flow cytometry, light microscopy, and immunofluorescence microscopy. After mixing normal RBC with platelet-rich plasma, discrete populations of RBC, platelets, and complexes of platelets attached to RBC were purified by fluorescence-activated cell sorting. Upon incubation of purified cells or platelets with HIV-1 followed by washing and co-incubation with CD4-positive peripheral blood mononuclear cells (PBMC, platelets, and platelet-RBC complexes, but not platelet-free RBC, caused infection of PBMC. Infection was prevented by pre-treating the platelet-RBC complexes with EDTA. Plasma and RBC (comprising a RBC/platelet-RBC mixture from chronically infected patients with low viral loads were also co-incubated with PBMC ex vivo to determine the presence of infectious HIV-1. All freshly isolated plasmas from the HIV-1-infected donors, obtained in the absence of anticoagulant, were noninfectious. Interestingly, the RBC from most of the patients caused cell-cell infection of PBMC that was prevented by stripping the RBC with EDTA. A monoclonal antibody to DC-SIGN partially inhibited cell-cell HIV-1 infection of PBMC by normal RBC pre-incubated with platelets and HIV-1. We conclude: (a platelet-free EDTA-free plasma from chronically infected HIV-1 patients, although containing viral RNA, is an environment that lacks detectable infectious HIV-1; (b platelets and platelet-RBC complexes, but not purified RBC, bind infectious HIV

  7. Purification and identification of the fusicoccin binding protein from oat root plasma membrane

    Science.gov (United States)

    de Boer, A. H.; Watson, B. A.; Cleland, R. E.

    1989-01-01

    Fusicoccin (FC), a fungal phytotoxin, stimulates the H(+) -ATPase located in the plasma membrane (PM) of higher plants. The first event in the reaction chain leading to enhanced H(+) -efflux seems to be the binding of FC to a FC-binding protein (FCBP) in the PM. We solubilized 90% of the FCBP from oat (Avena sativa L. cv Victory) root PM in an active form with 1% octyl-glucoside. The FCBP was stabilized by the presence of protease inhibitors. The FCBP was purified by affinity chromatography using FC-linked adipic acid dihydrazide agarose (FC-AADA). Upon elution with 8 molar urea, two major protein bands on sodium dodecyl sulfate-polyaerylamide gel electrophoresis with molecular weights of 29,700 and 31,000 were obtained. Successive chromatography on BBAB Bio-Gel A, hexyl agarose, and FC-AADA resulted in the same two bands when the FC-AADA was eluted with sodium dodecyl sulfate. A direct correlation was made between 3H-FC-binding activity and the presence of the two protein bands. The stoichiometry of the 29,700 and 31,000 molecular weight bands was 1:2. This suggests that the FCBP occurs in the native form as a heterotrimer with an apparent molecular weight of approximately 92,000.

  8. Increased plasma retinol binding protein 4 levels in patients with inflammatory cardiomyopathy.

    Science.gov (United States)

    Bobbert, Peter; Weithäuser, Alice; Andres, Janin; Bobbert, Thomas; Kühl, Uwe; Schultheiss, Heinz Peter; Rauch, Ursula; Skurk, Carsten

    2009-12-01

    Chronic heart failure (CHF) is associated with a higher risk for diabetes mellitus. Retinol binding protein 4 (RBP 4) is an adipose tissue-derived protein with pro-diabetogenic effects. A complete understanding of the association of CHF and insulin resistance remains elusive. The purpose of this study was to examine the relationship between CHF and diabetes mellitus. Plasma levels of RBP 4, insulin, and interleukins (IL) 2, 8, and 10, were assessed in patients with dilated cardiomyopathy (DCM, n = 53), dilated inflammatory cardiomyopathy (DCMi, n = 54), and controls (n = 20). In addition, a possible mechanism of RBP 4 regulation was examined in adipocytes in vitro. Plasma levels of RBP 4 and insulin were measured by a specific ELISA. Interleukin concentrations were obtained by multiplex ELISA. Cell culture with 3T3-L1 adipocytes was performed to measure RBP 4 mRNA expression after stimulation with IL-8. RBP 4 levels were significantly increased in patients with DCMi (52.95 +/- 20.42 microg/mL) compared with DCM (35.54 +/- 23.08 microg/mL) and the control group (27.3 +/- 18.51 microg/mL). RBP 4 was positively correlated with IL-8 (r=0.416, P < 0.05) in human plasma in patients with DCMi. Moreover, increased insulin resistance was observed in patients with DCMi compared with the control and DCM groups. In vitro, IL-8 induced a significant upregulation of RBP 4 mRNA expression in adipocytes. Elevated RBP 4 plasma concentrations, induced by IL-8, might be one mechanism leading to a higher incidence of diabetes in patients with DCMi.

  9. Informing the Human Plasma Protein Binding of Environmental Chemicals by Machine Learning in the Pharmaceutical Space: Applicability Domain and Limits of Predictability

    Science.gov (United States)

    The free fraction of a xenobiotic in plasma (Fub) is an important determinant of chemical adsorption, distribution, metabolism, elimination, and toxicity, yet experimental plasma protein binding data is scarce for environmentally relevant chemicals. The presented work explores th...

  10. Informing the Human Plasma Protein Binding of Environmental Chemicals by Machine Learning in the Pharmaceutical Space: Applicability Domain and Limits of Predictability

    Science.gov (United States)

    The free fraction of a xenobiotic in plasma (Fub) is an important determinant of chemical adsorption, distribution, metabolism, elimination, and toxicity, yet experimental plasma protein binding data is scarce for environmentally relevant chemicals. The presented work explores th...

  11. Isolation and characterization of gelatin-binding proteins from goat seminal plasma

    Directory of Open Access Journals (Sweden)

    Lazure Claude

    2003-04-01

    Full Text Available Abstract A family of proteins designated BSP-A1, BSP-A2, BSP-A3 and BSP-30 kDa (collectively called BSP proteins for Bovine Seminal Plasma proteins constitute the major protein fraction in the bull seminal plasma. These proteins interact with choline phospholipids on the sperm surface and play a role in the membrane stabilization (decapacitation and destabilization (capacitation process. Homologous proteins have been isolated from boar and stallion seminal plasma. In the current study we report the isolation and preliminary characterization of homologous proteins from goat seminal plasma. Frozen semen (-80°C was thawed and centrifuged to remove sperm. The proteins in the supernatant were precipitated by the addition of cold ethanol. The precipitates were dissolved in ammonium bicarbonate and lyophilised. The lyophilised proteins were dissolved in phosphate buffer and loaded onto a gelatin-agarose column, which was previously equilibrated with the same buffer. The column was successively washed with phosphate buffer, with phosphate buffer saline and with 0.5 M urea in phosphate buffer saline to remove unadsorbed proteins, and the adsorbed proteins were eluted with 5 M urea in phosphate buffer saline. Analysis of pooled, dialysed and lyophilised gelatin-agarose adsorbed protein fraction by SDS-PAGE indicated the presence of four protein bands that were designated GSP-14 kDa, GSP-15 kDa, GSP-20 kDa and GSP-22 kDa (GSP, Goat Seminal Plasma proteins. Heparin-affinity chromatography was then used for the separation of GSP-20 and -22 kDa from GSP-14 and -15 kDa. Finally, HPLC separation permitted further isolation of each one from the other. Amino acid sequence analysis of these proteins indicated that they are homologous to BSP proteins. In addition, these BSP homologs bind to hen's egg-yolk low-density lipoproteins. These results together with our previous data indicate that BSP family proteins are ubiquitous in mammalian seminal plasma, exist in

  12. Binding of plasma proteins to titanium dioxide nanotubes with different diameters

    Directory of Open Access Journals (Sweden)

    Kulkarni M

    2015-02-01

    Full Text Available Mukta Kulkarni,1,* Ajda Flašker,1,* Maruša Lokar,1 Katjuša Mrak-Poljšak,2 Anca Mazare,3 Andrej Artenjak,4 Saša Čučnik,2 Slavko Kralj,5 Aljaž Velikonja,1 Patrik Schmuki,3 Veronika Kralj-Iglič,6 Snezna Sodin-Semrl,2,7 Aleš Iglič11Laboratory of Biophysics, Faculty of Electrical Engineering, University of Ljubljana, Ljubljana, Slovenia; 2Department of Rheumatology, University Medical Centre Ljubljana, Ljubljana, Slovenia; 3Department of Materials Science and Engineering, University of Erlangen Nuremberg, Erlangen, Germany; 4Sandoz Biopharmaceuticals Mengeš, Lek Pharmaceuticals dd, Menges, Slovenia; 5Department for Materials Synthesis, Institute Jožef Stefan (IJS, Ljubljana, Slovenia; 6Faculty of Health Studies, University of Ljubljana, Ljubljana, Slovenia; 7Faculty of Mathematics, Natural Science and Information Technology, University of Primorska, Koper, Slovenia *These authors contributed equally to this workAbstract: Titanium and titanium alloys are considered to be one of the most applicable materials in medical devices because of their suitable properties, most importantly high corrosion resistance and the specific combination of strength with biocompatibility. In order to improve the biocompatibility of titanium surfaces, the current report initially focuses on specifying the topography of titanium dioxide (TiO2 nanotubes (NTs by electrochemical anodization. The zeta potential (ζ-potential of NTs showed a negative value and confirmed the agreement between the measured and theoretically predicted dependence of ζ-potential on salt concentration, whereby the absolute value of ζ-potential diminished with increasing salt concentrations. We investigated binding of various plasma proteins with different sizes and charges using the bicinchoninic acid assay and immunofluorescence microscopy. Results showed effective and comparatively higher protein binding to NTs with 100 nm diameters (compared to 50 or 15 nm. We also showed a dose

  13. The Association of Myosin IB with Actin Waves in Dictyostelium Requires Both the Plasma Membrane-Binding Site and Actin-Binding Region in the Myosin Tail

    Science.gov (United States)

    Brzeska, Hanna; Pridham, Kevin; Chery, Godefroy; Titus, Margaret A.; Korn, Edward D.

    2014-01-01

    F-actin structures and their distribution are important determinants of the dynamic shapes and functions of eukaryotic cells. Actin waves are F-actin formations that move along the ventral cell membrane driven by actin polymerization. Dictyostelium myosin IB is associated with actin waves but its role in the wave is unknown. Myosin IB is a monomeric, non-filamentous myosin with a globular head that binds to F-actin and has motor activity, and a non-helical tail comprising a basic region, a glycine-proline-glutamine-rich region and an SH3-domain. The basic region binds to acidic phospholipids in the plasma membrane through a short basic-hydrophobic site and the Gly-Pro-Gln region binds F-actin. In the current work we found that both the basic-hydrophobic site in the basic region and the Gly-Pro-Gln region of the tail are required for the association of myosin IB with actin waves. This is the first evidence that the Gly-Pro-Gln region is required for localization of myosin IB to a specific actin structure in situ. The head is not required for myosin IB association with actin waves but binding of the head to F-actin strengthens the association of myosin IB with waves and stabilizes waves. Neither the SH3-domain nor motor activity is required for association of myosin IB with actin waves. We conclude that myosin IB contributes to anchoring actin waves to the plasma membranes by binding of the basic-hydrophobic site to acidic phospholipids in the plasma membrane and binding of the Gly-Pro-Gln region to F-actin in the wave. PMID:24747353

  14. Pretreatment of plasma samples by a novel hollow fiber centrifugal ultrafiltration technique for the determination of plasma protein binding of three coumarins using acetone as protein binding releasing agent.

    Science.gov (United States)

    Li, Junmei; Shi, Qingwen; Jiang, Ye; Liu, Yan

    2015-09-15

    A novel and practical sample pretreatment method based on hollow fiber centrifugal ultrafiltration (HFCF-UF) was developed to determine plasma protein binding by using HPLC. The samples for analyzing unbound and total concentrations could be prepared in parallel simultaneously by the same device. It only required centrifugation for a short time and the filtrate could be injected directly for HPLC analysis without further treatment. Coumarins were selected as the model drugs. Acetone was chosen as the releasing agent to free the binding drug from the drug-protein complex for the total drug concentration determination. Non-specific bindings (NSBs) between the analytes and hollow fiber membrane materials were investigated. The type and volume of protein binding releaser were optimized. Additionally, centrifugal speed and centrifugal time were considered. Under the optimized conditions, the absolute recovery rates of the unbound and total concentrations were in the range of 97.5-100.9% for the three analytes. The limits of detection were in the range of 0.0135-0.0667μgmL(-1). In vitro plasma protein binding of the three coumarins was determined at three concentrations using the validated method and the relative standard deviations (RSDs) were less than 3.4%. Compared with traditional method, the HFCF-UF method is simple to run, no specialized equipment requirement and is a more accurate plasma pretreatment procedure with almost excellent drug-protein binding equilibrium. Therefore, this method can be applied to determine the plasma protein binding in clinical practice. It also provides a reliable alternative for accurate monitoring of unbound or total drug concentration in therapeutic drug monitoring (TDM).

  15. Characterization of the microheterogeneity of transthyretin in plasma and urine using SELDI-TOF-MS immunoassay

    Directory of Open Access Journals (Sweden)

    Raila Jens

    2004-09-01

    Full Text Available Abstract Background It has been shown that transthyretin (TTR exists in different molecular variants. Besides point mutations associated with different diseases such as amyloidosis, other posttranslational modifications occur that might be of diagnostic interest. Results TTR levels as determined by ELISA in plasma and urine of healthy individuals were 489 ± 155 μg/ml plasma and 46 ± 24 ng/g creatinine, respectively. Average levels in urine of pregnant women were 45 ± 65 μg/g creatinine. The molecular heterogeneity of TTR was analyzed using a high-throughput mass spectrometric immunoassay system. TTR was extracted from plasma or urine onto an antibody-coated (via protein A affinity chip surface (PS20 using the surface-enhanced laser desorption/ionization (SELDI technique. Subsequently samples were subjected to time-of-flight mass spectrometry (TOF-MS. In healthy individuals, TTR in plasma occurred rather consistently in two variants of 13732 ± 12 and 13851 ± 9 Da for the native and S-cysteinylated forms and at a smaller signal of 14043 ± 17 Da for the S-glutathionylated form. In urine of pregnant women, various signals were observed with a dominant signal at 13736 ± 10 Da and a varying number of smaller immunoreactive fragments. These fragments are possibly the consequence of metabolism in plasma or kidney. Conclusion This chip-based approach represents a rapid and accurate method to characterize the molecular variants of TTR including protein or peptide fragments which are either related to TTR or have resulted from its catabolism. These molecular variants may be of diagnostic importance as alternative or novel biomarkers due to their predominant relation to the TTR metabolism both in healthy and diseased individuals.

  16. Cyanide binding to human plasma heme-hemopexin: A comparative study

    Energy Technology Data Exchange (ETDEWEB)

    Ascenzi, Paolo, E-mail: ascenzi@uniroma3.it [Laboratorio Interdipartimentale di Microscopia Elettronica, Universita Roma Tre, Roma (Italy); Istituto Nazionale di Biostrutture e Biosistemi, Roma (Italy); Leboffe, Loris [Istituto Nazionale di Biostrutture e Biosistemi, Roma (Italy); Polticelli, Fabio [Dipartimento di Biologia, Universita Roma Tre, Roma (Italy)

    2012-11-16

    Highlights: Black-Right-Pointing-Pointer Cyanide binding to ferric HHPX-heme-Fe. Black-Right-Pointing-Pointer Cyanide binding to ferrous HHPX-heme-Fe. Black-Right-Pointing-Pointer Dithionite-mediated reduction of ferric HHPX-heme-Fe-cyanide. Black-Right-Pointing-Pointer Cyanide binding to HHPX-heme-Fe is limited by ligand deprotonation. Black-Right-Pointing-Pointer Cyanide dissociation from HHPX-heme-Fe-cyanide is limited by ligand protonation. -- Abstract: Hemopexin (HPX) displays a pivotal role in heme scavenging and delivery to the liver. In turn, heme-Fe-hemopexin (HPX-heme-Fe) displays heme-based spectroscopic and reactivity properties. Here, kinetics and thermodynamics of cyanide binding to ferric and ferrous hexa-coordinate human plasma HPX-heme-Fe (HHPX-heme-Fe(III) and HHPX-heme-Fe(II), respectively), and for the dithionite-mediated reduction of the HHPX-heme-Fe(III)-cyanide complex, at pH 7.4 and 20.0 Degree-Sign C, are reported. Values of thermodynamic and kinetic parameters for cyanide binding to HHPX-heme-Fe(III) and HHPX-heme-Fe(II) are K = (4.1 {+-} 0.4) Multiplication-Sign 10{sup -6} M, k{sub on} = (6.9 {+-} 0.5) Multiplication-Sign 10{sup 1} M{sup -1} s{sup -1}, and k{sub off} = 2.8 Multiplication-Sign 10{sup -4} s{sup -1}; and H = (6 {+-} 1) Multiplication-Sign 10{sup -1} M, h{sub on} = 1.2 Multiplication-Sign 10{sup -1} M{sup -1} s{sup -1}, and h{sub off} = (7.1 {+-} 0.8) Multiplication-Sign 10{sup -2} s{sup -1}, respectively. The value of the rate constant for the dithionite-mediated reduction of the HHPX-heme-Fe(III)-cyanide complex is l = 8.9 {+-} 0.8 M{sup -1/2} s{sup -1}. HHPX-heme-Fe reactivity is modulated by proton acceptor/donor amino acid residue(s) (e.g., His236) assisting the deprotonation and protonation of the incoming and outgoing ligand, respectively.

  17. Plasma lactate, GH and GH-binding protein levels in exercise following BCAA supplementation in athletes.

    Science.gov (United States)

    De Palo, E F; Gatti, R; Cappellin, E; Schiraldi, C; De Palo, C B; Spinella, P

    2001-01-01

    Branched chain amino acids (BCAA) stimulate protein synthesis, and growth hormone (GH) is a mediator in this process. A pre-exercise BCAA ingestion increases muscle BCAA uptake and use. Therefore after one month of chronic BCAA treatment (0.2 gkg(-1) of body weight), the effects of a pre-exercise oral supplementation of BCAA (9.64 g) on the plasma lactate (La) were examined in triathletes, before and after 60 min of physical exercise (75% of VO2 max). The plasma levels of GH (pGH) and of growth hormone binding protein (pGHBP) were also studied. The end-exercise La of each athlete was higher than basal. Furthermore, after the chronic BCAA treatment, these end-exercise levels were lower than before this treatment (8.6+/-0.8 mmol L(-1) after vs 12.8+/-1.0 mmol L(-1) before treatment; p BCAA chronic treatment, this end-exercise pGHBP was 738+/-85 pmol L(-1) before vs 1691+/-555 pmol L(-1) after. pGH/pGHBP ratio was unchanged in each athlete and between the groups, but a tendency to increase was observed at end-exercise. The lower La at the end of an intense muscular exercise may reflect an improvement of BCAA use, due to the BCAA chronic treatment. The chronic BCAA effects on pGH and pGHBP might suggest an improvement of muscle activity through protein synthesis.

  18. Aggregation analysis of Con A binding proteins of human seminal plasma: a dynamic light scattering study.

    Science.gov (United States)

    Tomar, Anil Kumar; Sooch, Balwinder Singh; Singh, Sarman; Yadav, Savita

    2013-02-01

    Concanavalin A (Con A) binding fraction of human seminal plasma is vital as it shows decapacitating activity and contains proteins which have critical roles in fertility related processes. Con A binding proteins were isolated by lectin affinity chromatography. These proteins form high molecular weight aggregates at near physiological pH (7.0) as inferred by gel filtration. Aggregation analysis was performed by dynamic light scattering (DLS). DLS analysis was also performed at different pH values and in presence of various additives including NaCl, EDTA, cholesterol and sugars, such as d-glucose, d-fructose and d-mannose to identify their effect on aggregation size. The results indicate that degree of aggregation was highly reduced in presence of d-fructose, EDTA and at lower and higher pH values as depicted by lowering of hydrodynamic radii. This aggregation behaviour might be decisive for fertility related events with a suggestive role towards inhibition of premature capacitation.

  19. Lectin receptors on the plasma membrane of soybean cells. Binding and lateral diffusion of lectins.

    Science.gov (United States)

    Metcalf, T N; Wang, J L; Schubert, K R; Schindler, M

    1983-08-02

    Protoplasts prepared from suspension cultures of root cells of Glycine max (SB-1 cell line) bound soybean agglutinin (SBA), concanavalin A (Con A), and wheat germ agglutinin (WGA). Binding studies carried out with 125I-labeled SBA, Con A, and WGA showed that these interactions were saturable and specific. Fluorescence microscopy demonstrated uniform membrane labeling. The mobility of the lectin-receptor complexes was measured by fluorescence redistribution after photobleaching. The diffusion constants (D) for SBA and Con A were 5 X 10(-11) and 7 X 10(-11) cm2/s, respectively. In contrast, WGA yielded a diffusion constant of 3 X 10(-10) cm2/s. Pretreatment of the protoplasts with either SBA or Con A resulted in a 6-fold reduction in the mobility of WGA (D congruent to 5 X 10(-11) cm2/s). The results suggest that the binding of SBA or Con A may lead to alterations of the soybean plasma membrane which, in turn, may restrict the mobility of other receptors.

  20. Plasma Protein Binding of Anisomelic Acid: Spectroscopy and Molecular Dynamic Simulations.

    Science.gov (United States)

    Senthilkumar, Rajendran; Marimuthu, Parthiban; Paul, Preethy; Manojkumar, Yesaiyan; Arunachalam, Sankaralingam; Eriksson, John E; Johnson, Mark S

    2016-12-27

    Anisomelic acid (AA) is a macrocyclic cembranolide compound extracted from Anisomeles herbal species. Recently, we have shown that AA possesses both anticancer and antiviral activity. However, to date, the plasma protein binding properties of AA are unknown. Here, we describe the molecular interactions of AA with two serum proteins, human serum albumin (HSA) and bovine serum albumin (BSA), adopting multiple physicochemical methods. Besides, molecular docking and dynamics simulations were performed to predict the interaction mode and the dynamic behavior of AA with HSA and BSA. The experimental results revealed that hydrophobic forces play a significant part in the interaction of AA to HSA and BSA. The outcomes of the principal components analysis (PCA) of the poses based on root-mean-squared distances showed less variation in AA-HSA, opposed to what is seen for BSA-AA. Furthermore, binding free energies estimated for AA-HSA and AA-BSA complexes at different temperatures (298, 303, 308, and 313 K) based on molecular mechanics-generalized Born surface area (MMGBSA) approaches were well correlated with our experimental results.

  1. Plasma mannose-binding lectin is stimulated by PPARα in humans.

    Science.gov (United States)

    Rakhshandehroo, Maryam; Stienstra, Rinke; de Wit, Nicole J; Bragt, Marjolijn C E; Haluzik, Martin; Mensink, Ronald P; Müller, Michael; Kersten, Sander

    2012-03-01

    The peroxisome proliferator activated receptor-α (PPARα) is a major transcriptional regulator of lipid metabolism in liver and represents the molecular target for hypolipidemic fibrate drugs. Effects of PPARα on lipid metabolism are partially mediated by circulating proteins such as FGF21 and ANGPTL4. The present study was undertaken to screen for and identify circulating proteins produced by human liver that are under the control of PPARα. Toward that aim, primary human hepatocytes were treated with the synthetic PPARα agonist Wy-14643 and whole genome expression data selected for secreted proteins. Expression of FGF21, ANGPTL4, and mannose-binding lectin (MBL), a soluble mediator of innate immunity and primary component of the lectin branch of the complement system, was markedly upregulated by Wy-14643 in primary human hepatocytes. Mice express two MBL isomers, Mbl1 and Mbl2. Mbl1 mRNA was weakly induced by Wy-14643 in primary mouse hepatocytes and remained unaltered by Wy-14643 in mouse liver. Mbl2 mRNA was unchanged by Wy-14643 in primary mouse hepatocytes and was strongly reduced by Wy-14643 in mouse liver. Remarkably, plasma Mbl1 levels were increased by chronic PPARα activation in lean and obese mice. Importantly, in two independent clinical trials, treatment with the PPARα agonist fenofibrate at 200 mg/day for 6 wk and 3 mo increased plasma MBL levels by 73 (P = 0.0016) and 86% (P = 0.017), respectively. It is concluded that hepatocyte gene expression and plasma levels of MBL are stimulated by PPARα and fenofibrate in humans, linking PPARα to regulation of innate immunity and complement activation in humans and suggesting a possible role of MBL in lipid metabolism.

  2. Genome wide association identifies common variants at the SERPINA6/SERPINA1 locus influencing plasma cortisol and corticosteroid binding globulin.

    Directory of Open Access Journals (Sweden)

    Jennifer L Bolton

    2014-07-01

    Full Text Available Variation in plasma levels of cortisol, an essential hormone in the stress response, is associated in population-based studies with cardio-metabolic, inflammatory and neuro-cognitive traits and diseases. Heritability of plasma cortisol is estimated at 30-60% but no common genetic contribution has been identified. The CORtisol NETwork (CORNET consortium undertook genome wide association meta-analysis for plasma cortisol in 12,597 Caucasian participants, replicated in 2,795 participants. The results indicate that <1% of variance in plasma cortisol is accounted for by genetic variation in a single region of chromosome 14. This locus spans SERPINA6, encoding corticosteroid binding globulin (CBG, the major cortisol-binding protein in plasma, and SERPINA1, encoding α1-antitrypsin (which inhibits cleavage of the reactive centre loop that releases cortisol from CBG. Three partially independent signals were identified within the region, represented by common SNPs; detailed biochemical investigation in a nested sub-cohort showed all these SNPs were associated with variation in total cortisol binding activity in plasma, but some variants influenced total CBG concentrations while the top hit (rs12589136 influenced the immunoreactivity of the reactive centre loop of CBG. Exome chip and 1000 Genomes imputation analysis of this locus in the CROATIA-Korcula cohort identified missense mutations in SERPINA6 and SERPINA1 that did not account for the effects of common variants. These findings reveal a novel common genetic source of variation in binding of cortisol by CBG, and reinforce the key role of CBG in determining plasma cortisol levels. In turn this genetic variation may contribute to cortisol-associated degenerative diseases.

  3. Plasma-treated polystyrene film that enhances binding efficiency for sensitive and label-free protein biosensing

    Science.gov (United States)

    Guo, Bihong; Li, Shaopeng; Song, Lusheng; Yang, Mo; Zhou, Wenfei; Tyagi, Deependra; Zhu, Jinsong

    2015-08-01

    A plasma-treated ultrathin polystyrene (PS) film surface was explored as a simple, robust, and low-cost surface chemistry solution for protein biosensing applications. This surface could dramatically improve the binding efficiency of the protein-protein interactions, which is defined as the binding signal per immobilized ligand. The PS-modified protein biosensor was readily fabricated by spin coating and plasma treatment. Various parameters for fabrication, including the concentration of the PS solution, rate of spin coating, and duration of plasma treatment, were systematically optimized based on the improvement of fluorescence signal yielded by the microfluidic network-aided fluorescence immunoassay. The performance of the label-free protein detection on the optimized surfaces was further evaluated by surface plasmon resonance imaging (SPRi). PS surfaces with optimal fabrication parameters exhibited up to an 620% enhancement of the protein binding response and approximately 210% of the protein binding per immobilized protein ligand compared with a self-assembled monolayer (SAM) surface of 11-mercapto undecanoic acid (MUA). The relationship between the fabrication parameters used and changes to the surface chemistry and the morphological properties were characterized with atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS), and Fourier transform infrared spectroscopy (FTIR). It was revealed that the morphological changes observed in the plasma-treated PS film were the dominant factor for the improvement of the protein bioassay performance, rather than the chemical changes.

  4. Low prevalence of antibodies and other plasma factors binding to CC chemokines and IL-2 in HIV-positive patients

    DEFF Research Database (Denmark)

    Meyer, C N; Svenson, M; Larsen, Carsten Schade;

    2000-01-01

    Neutralizing cytokine antibodies are found in healthy and diseased individuals, including patients treated with recombinant cytokines. Identification of CCR-5 as co-receptor for HIV has focused interest on CC chemokines and their potential therapeutic use. Chemokine-binding components in plasma...... of HIV-infected patients were therefore assessed by radioimmunoassay and radioreceptor assay. IgG from 4/505 HIV patients and 9/2000 healthy controls (p>0.05) bound rMIP-1alpha and rMIP-1beta, but not rRANTES. No other plasma factors bound the chemokines. The antibodies inhibited receptor binding of both...... chemokines. There was no association between presence of antibodies and disease stage or HIV progression rate. Three of 11 patients treated with rIL-2 developed IgG antibodies suppressing cellular binding and growth promotion of rIL-2. Hence, circulating factors, including antibodies MIP-1alpha/MIP-1beta...

  5. Low prevalence of antibodies and other plasma factors binding to CC chemokines and IL-2 in HIV-positive patients

    DEFF Research Database (Denmark)

    Meyer, C N; Svenson, M; Schade Larsen, C

    2000-01-01

    chemokines. There was no association between presence of antibodies and disease stage or HIV progression rate. Three of 11 patients treated with rIL-2 developed IgG antibodies suppressing cellular binding and growth promotion of rIL-2. Hence, circulating factors, including antibodies MIP-1alpha/MIP-1beta......Neutralizing cytokine antibodies are found in healthy and diseased individuals, including patients treated with recombinant cytokines. Identification of CCR-5 as co-receptor for HIV has focused interest on CC chemokines and their potential therapeutic use. Chemokine-binding components in plasma...... of HIV-infected patients were therefore assessed by radioimmunoassay and radioreceptor assay. IgG from 4/505 HIV patients and 9/2000 healthy controls (p>0.05) bound rMIP-1alpha and rMIP-1beta, but not rRANTES. No other plasma factors bound the chemokines. The antibodies inhibited receptor binding of both...

  6. Plasma-treated polystyrene film that enhances binding efficiency for sensitive and label-free protein biosensing

    Energy Technology Data Exchange (ETDEWEB)

    Guo, Bihong [National Center for NanoScience and Technology, No. 11 Beiyitiao, Zhongguancun, Beijing 100190 (China); Li, Shaopeng [National Center for NanoScience and Technology, No. 11 Beiyitiao, Zhongguancun, Beijing 100190 (China); Department of Chemistry, Tsinghua University, Beijing 100084 (China); Song, Lusheng [National Center for NanoScience and Technology, No. 11 Beiyitiao, Zhongguancun, Beijing 100190 (China); Yang, Mo; Zhou, Wenfei; Tyagi, Deependra [National Center for NanoScience and Technology, No. 11 Beiyitiao, Zhongguancun, Beijing 100190 (China); University of Chinese Academy of Sciences, Yuquan Rd., 19(A), Beijing 100049 (China); Zhu, Jinsong, E-mail: jizhu88@gmail.com [National Center for NanoScience and Technology, No. 11 Beiyitiao, Zhongguancun, Beijing 100190 (China)

    2015-08-01

    Highlights: • A simple and robust plasma-treated ultrathin polystyrene film surface was developed for protein biosensing. • The surface was optimized by evaluating up to 120 types of fabrication parameters with high-throughput analytical methods. • The optimized surface showed a 620% improvement of the protein detection signal and 210% protein binding per immobilized protein ligand compared with a self-assembled monolayer surface. - Abstract: A plasma-treated ultrathin polystyrene (PS) film surface was explored as a simple, robust, and low-cost surface chemistry solution for protein biosensing applications. This surface could dramatically improve the binding efficiency of the protein–protein interactions, which is defined as the binding signal per immobilized ligand. The PS-modified protein biosensor was readily fabricated by spin coating and plasma treatment. Various parameters for fabrication, including the concentration of the PS solution, rate of spin coating, and duration of plasma treatment, were systematically optimized based on the improvement of fluorescence signal yielded by the microfluidic network-aided fluorescence immunoassay. The performance of the label-free protein detection on the optimized surfaces was further evaluated by surface plasmon resonance imaging (SPRi). PS surfaces with optimal fabrication parameters exhibited up to an 620% enhancement of the protein binding response and approximately 210% of the protein binding per immobilized protein ligand compared with a self-assembled monolayer (SAM) surface of 11-mercapto undecanoic acid (MUA). The relationship between the fabrication parameters used and changes to the surface chemistry and the morphological properties were characterized with atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS), and Fourier transform infrared spectroscopy (FTIR). It was revealed that the morphological changes observed in the plasma-treated PS film were the dominant factor for the

  7. SPM analysis of parametric (R)-[11C]PK11195 binding images: plasma input versus reference tissue parametric methods.

    Science.gov (United States)

    Schuitemaker, Alie; van Berckel, Bart N M; Kropholler, Marc A; Veltman, Dick J; Scheltens, Philip; Jonker, Cees; Lammertsma, Adriaan A; Boellaard, Ronald

    2007-05-01

    (R)-[11C]PK11195 has been used for quantifying cerebral microglial activation in vivo. In previous studies, both plasma input and reference tissue methods have been used, usually in combination with a region of interest (ROI) approach. Definition of ROIs, however, can be labourious and prone to interobserver variation. In addition, results are only obtained for predefined areas and (unexpected) signals in undefined areas may be missed. On the other hand, standard pharmacokinetic models are too sensitive to noise to calculate (R)-[11C]PK11195 binding on a voxel-by-voxel basis. Linearised versions of both plasma input and reference tissue models have been described, and these are more suitable for parametric imaging. The purpose of this study was to compare the performance of these plasma input and reference tissue parametric methods on the outcome of statistical parametric mapping (SPM) analysis of (R)-[11C]PK11195 binding. Dynamic (R)-[11C]PK11195 PET scans with arterial blood sampling were performed in 7 younger and 11 elderly healthy subjects. Parametric images of volume of distribution (Vd) and binding potential (BP) were generated using linearised versions of plasma input (Logan) and reference tissue (Reference Parametric Mapping) models. Images were compared at the group level using SPM with a two-sample t-test per voxel, both with and without proportional scaling. Parametric BP images without scaling provided the most sensitive framework for determining differences in (R)-[11C]PK11195 binding between younger and elderly subjects. Vd images could only demonstrate differences in (R)-[11C]PK11195 binding when analysed with proportional scaling due to intersubject variation in K1/k2 (blood-brain barrier transport and non-specific binding).

  8. Characterization of gonadotropin binding sites in the intracellular organelles of bovine corpora lutea and comparison with plasma membrane sites.

    Science.gov (United States)

    Rao, C V; Mitra, S; Carman, F R

    1981-03-25

    The specific binding of 125I-human choriogonadotropin (hCG) to plasma membranes, nuclear membranes, lysosomes, rough endoplasmic reticulum, heavy golgi, and medium and light golgi of bovine corpora lutea was dependent on the amount of protein, 125I-hCG concentration and incubation time. The bound hormone in all the organelles was able to rebind to fresh corresponding organelles. Scatchard analysis revealed a homogenous population of gonadotropin binding sites in plasma membrane, rough endoplasmic reticulum, heavy golgi, and medium and light golgi, whose binding affinities (Kd = 8.6-11.0 X 10(-11) M) were similar but whose number of available gonadotropin binding sites varied. Scatchard analyses of nuclear membranes and lysosome binding, on the other hand, were heterogenous (Nuclear membranes, 11 and 23 X 10(-11) M lysosomes, 3.4 and 130 X 10(-11) M). The rate constants for association (5.9 to 12.1 X 10(6) M-1 S-1) and dissociation (7.4 to 9.0 X 10(-4) S-1) were similar among different subcellular organelles except for nuclear membranes and lysosomes, where rate constants for association were significantly lower. The ligand binding specificity, lower effectiveness of human luteinizing hormone as compared to hCG in competition, the optimal pH, the lack of ionic requirements for binding, and the molecular size of 125I-hCG-gonadotropin binding site complexes solubilized from various intracellular organelles were similar to those observed for plasma membranes. Numerous differences were also observed between intracellular organelles and plasma membranes as well as among intracellular organelles themselves with respect to binding losses due to exposure to low and high pH values, di- and monovalent cations, increasing preincubation temperatures, and a variety of enzymes and protein reagents. The possible reasons for these similarities as well as differences observed are discussed. The differences are viewed as an additional indication that contamination cannot solely

  9. Interaction of gold and silver nanoparticles with human plasma: Analysis of protein corona reveals specific binding patterns.

    Science.gov (United States)

    Lai, Wenjia; Wang, Qingsong; Li, Lumeng; Hu, Zhiyuan; Chen, Jiankui; Fang, Qiaojun

    2017-04-01

    Determining how nanomaterials interact with plasma will assist in understanding their effects on the biological system. This work presents a systematic study of the protein corona formed from human plasma on 20nm silver and gold nanoparticles with three different surface modifications, including positive and negative surface charges. The results show that all nanoparticles, even those with positive surface modifications, acquire negative charges after interacting with plasma. Approximately 300 proteins are identified on the coronas, while 99 are commonly found on each nanomaterial. The 20 most abundant proteins account for over 80% of the total proteins abundance. Remarkably, the surface charge and core of the nanoparticles, as well as the isoelectric point of the plasma proteins, are found to play significant roles in determining the nanoparticle coronas. Albumin and globulins are present at levels of less than 2% on these nanoparticle coronas. Fibrinogen, which presents in the plasma but not in the serum, preferably binds to negatively charged gold nanoparticles. These observations demonstrate the specific plasma protein binding pattern of silver and gold nanoparticles, as well as the importance of the surface charge and core in determining the protein corona compositions. The potential downstream biological impacts of the corona proteins were also investigated.

  10. Radioreceptor assay to study the affinity of benzodiazepines and their receptor binding activity in human plasma including their active metabolites

    Energy Technology Data Exchange (ETDEWEB)

    Dorow, R.G.; Seidler, J.; Schneider, H.H. (Schering A.G., Berlin (Germany, F.R.))

    1982-04-01

    A radioreceptor assay has been established to measure the receptor affinities of numerous benzodiazepines in clinical use. The time course of receptor binding activity was studied by this method in the plasma of eight healthy subjects randomly treated with 1mg lormetazepam (Noctamid(R)), 2mg flunitrazepam (Rohypnol(R)), and 10mg diazepam (Valium(R)), and placebo on a cross-over basis. Blood samples were collected up to 154h after treatment. Receptor affinities of numerous benzodiazepines in vitro show good correlation with therapeutic human doses (r=0.96) and may be predictive of drug potency in man. Mean peak plasma levels of lormetazepam binding equivalents were 4.8+-1 ng/ml at 2h after lormetazepam, 7.2+-1.8 ng/ml at 8h after flunitrazepam, and 17.9+-2.7 ng/ml at 15h after diazepam. Plasma elimination halflives of benzodiazepine binding equivalents were 9.3, 23 and 63h, respectively. Slow elimination of benzodiazepine binding equivalents following flunitrazepam and diazepam may be due to persistent active metabolites.

  11. Androgen Receptor Localizes to Plasma Membrane by Binding to Caveolin-1 in Mouse Sertoli Cells.

    Science.gov (United States)

    Deng, Qiong; Wu, Yong; Zhang, Zeng; Wang, Yue; Li, Minghua; Liang, Hui; Gui, Yaoting

    2017-01-01

    The nonclassical androgen signaling pathway translates signals into alterations in cellular function within minutes, and this action is proposed to be mediated by an androgen receptor (AR) localized to the plasma membrane. This study was designed to determine the mechanism underlying the membrane association of androgen receptor in TM4 cells, a mouse Sertoli cell line. Western blot analysis indicated testosterone-induced AR translocation to the cell membrane. Data from coimmunoprecipitation indicated that AR is associated with caveolin-1, and testosterone enhanced this association. Knockdown of caveolin-1 by shRNA decreased the amount of AR localized to membrane fraction and prevented AR membrane trafficking after being exposed to testosterone at physiological concentration. The palmitoylation inhibitor 2-bromopalmitate decreased AR membrane localization in basal condition and completely blocked testosterone-induced AR translocation to membrane fraction. These data suggested that AR localized to membrane fraction by binding with caveolin-1 through palmitoylation of the cysteine residue. This study provided a new evidence for AR membrane localization and its application for clarifying the nonclassical signaling pathway of androgens.

  12. Androgen Receptor Localizes to Plasma Membrane by Binding to Caveolin-1 in Mouse Sertoli Cells

    Directory of Open Access Journals (Sweden)

    Qiong Deng

    2017-01-01

    Full Text Available The nonclassical androgen signaling pathway translates signals into alterations in cellular function within minutes, and this action is proposed to be mediated by an androgen receptor (AR localized to the plasma membrane. This study was designed to determine the mechanism underlying the membrane association of androgen receptor in TM4 cells, a mouse Sertoli cell line. Western blot analysis indicated testosterone-induced AR translocation to the cell membrane. Data from coimmunoprecipitation indicated that AR is associated with caveolin-1, and testosterone enhanced this association. Knockdown of caveolin-1 by shRNA decreased the amount of AR localized to membrane fraction and prevented AR membrane trafficking after being exposed to testosterone at physiological concentration. The palmitoylation inhibitor 2-bromopalmitate decreased AR membrane localization in basal condition and completely blocked testosterone-induced AR translocation to membrane fraction. These data suggested that AR localized to membrane fraction by binding with caveolin-1 through palmitoylation of the cysteine residue. This study provided a new evidence for AR membrane localization and its application for clarifying the nonclassical signaling pathway of androgens.

  13. Closure Report for Corrective Action Unit 412: Clean Slate I Plutonium Dispersion (TTR) Tonopah Test Range, Nevada, Revision 0

    Energy Technology Data Exchange (ETDEWEB)

    Matthews, Patrick [Navarro, Las Vegas, NV (United States)

    2016-08-22

    This Closure Report (CR) presents information supporting the clean closure of Corrective Action Unit (CAU) 412: Clean Slate I Plutonium Dispersion (TTR), located on the Tonopah Test Range, Nevada. CAU 412 consists of a release of radionuclides to the surrounding soil from a storage–transportation test conducted on May 25, 1963. Corrective action investigation (CAI) activities were performed in April and May 2015, as set forth in the Streamlined Approach for Environmental Restoration (SAFER) Plan for Corrective Action Unit 412: Clean Slate I Plutonium Dispersion (TTR), Tonopah Test Range, Nevada; and in accordance with the Soils Activity Quality Assurance Plan. The purpose of the CAI was to fulfill data needs as defined during the data quality objectives process. The CAU 412 dataset of investigation results was evaluated based on a data quality assessment. This assessment demonstrated the dataset is complete and acceptable for use in fulfilling the data needs identified by the data quality objectives process. This CR provides documentation and justification for the clean closure of CAU 412 under the FFACO without further corrective action. This justification is based on historical knowledge of the site, previous site investigations, implementation of the 1997 interim corrective action, and the results of the CAI. The corrective action of clean closure was confirmed as appropriate for closure of CAU 412 based on achievement of the following closure objectives: Radiological contamination at the site is less than the final action level using the ground troops exposure scenario (i.e., the radiological dose is less than the final action level): Removable alpha contamination is less than the high contamination area criterion: No potential source material is present at the site, and any impacted soil associated with potential source material has been removed so that remaining soil contains contaminants at concentrations less than the final action levels: and There is

  14. Closure Report for Corrective Action Unit 412: Clean Slate I Plutonium Dispersion (TTR) Tonopah Test Range, Nevada, Revision 0

    Energy Technology Data Exchange (ETDEWEB)

    Matthews, Patrick [Navarro, Las Vegas, NV (United States)

    2016-08-01

    This Closure Report (CR) presents information supporting the clean closure of Corrective Action Unit (CAU) 412: Clean Slate I Plutonium Dispersion (TTR), located on the Tonopah Test Range, Nevada. CAU 412 consists of a release of radionuclides to the surrounding soil from a storage–transportation test conducted on May 25, 1963. Corrective action investigation (CAI) activities were performed in April and May 2015, as set forth in the Streamlined Approach for Environmental Restoration (SAFER) Plan for Corrective Action Unit 412: Clean Slate I Plutonium Dispersion (TTR), Tonopah Test Range, Nevada; and in accordance with the Soils Activity Quality Assurance Plan. The purpose of the CAI was to fulfill data needs as defined during the data quality objectives process. The CAU 412 dataset of investigation results was evaluated based on a data quality assessment. This assessment demonstrated the dataset is complete and acceptable for use in fulfilling the data needs identified by the data quality objectives process. This CR provides documentation and justification for the clean closure of CAU 412 under the FFACO without further corrective action. This justification is based on historical knowledge of the site, previous site investigations, implementation of the 1997 interim corrective action, and the results of the CAI. The corrective action of clean closure was confirmed as appropriate for closure of CAU 412 based on achievement of the following closure objectives: • Radiological contamination at the site is less than the final action level using the ground troops exposure scenario (i.e., the radiological dose is less than the final action level). • Removable alpha contamination is less than the high contamination area criterion. • No potential source material is present at the site, and any impacted soil associated with potential source material has been removed so that remaining soil contains contaminants at concentrations less than the final action levels.

  15. 牡荆素血浆蛋白结合率的测定%Determination of plasma protein binding rate of vitexin

    Institute of Scientific and Technical Information of China (English)

    童成亮; 吴小英

    2012-01-01

    目的:建立血浆中牡荆素浓度的分析方法,测定牡荆素血浆蛋白结合率.方法:采用平衡透析法透析,利用HPLC测定血浆中牡荆素浓度,计算牡荆素与人血浆蛋白结合率,并比较大鼠和人血浆中牡荆素的血浆蛋白结合率.结果:在2 ~16 mg·L-1,药物浓度对血浆蛋白结合率无显著影响,但牡荆素在大鼠和人的血浆蛋白结合率存在显著性差异,牡荆素与人的血浆蛋白结合率高于与大鼠血浆蛋白结合率.结论:牡荆素与血浆蛋白具有较强的结合.%Objective: To establish an analytical method on vitexin concentration in plasma to determine the plasma protein binding rate of vitexin. Method: The equilibrium dialysis method and HPLC were adopted to determine vitexin concentration in plasma , calculate human plasma protein binding rate of vitexin and compare rat and human plasma protein binding rates of vitexin. Result: At 2-16 mg · L-1 , there was no significant difference in the plasma protein binding rate. But the human plasma protein binding rate of vitexin was higher than its rat plasma protein binding rate, indicating a significant difference in rat and human plasma protein binding rates of vitexin. Conclusion: Vitexin has a higher protein binding rate with both rat plasma and human plasma.

  16. Regulation of IGF binding protein proteolysis by pregnancy-associated plasma protein-ARegulation of IGF binding protein proteolysis by pregnancy-associated plasma protein-A

    DEFF Research Database (Denmark)

    Gaidamauskas, Ervinas

    During his PhD studies, Ervinas Gaidamauskas researched the proteins pregnancy-associated plasma protein-A (PAPP-A) and its homologue PAPP-A2 in vitro. As suggested by its name, PAPP-A plays an important role in pregnancy and fetal development. Additionally, recent studies indicate a newly...

  17. Regulation of IGF binding protein proteolysis by pregnancy-associated plasma protein-ARegulation of IGF binding protein proteolysis by pregnancy-associated plasma protein-A

    DEFF Research Database (Denmark)

    Gaidamauskas, Ervinas

    During his PhD studies, Ervinas Gaidamauskas researched the proteins pregnancy-associated plasma protein-A (PAPP-A) and its homologue PAPP-A2 in vitro. As suggested by its name, PAPP-A plays an important role in pregnancy and fetal development. Additionally, recent studies indicate a newly...

  18. Identification of CD4-Binding Site Dependent Plasma Neutralizing Antibodies in an HIV-1 Infected Indian Individual.

    Science.gov (United States)

    Khan, Lubina; Makhdoomi, Muzamil Ashraf; Kumar, Sanjeev; Nair, Ambili; Andrabi, Raiees; Clark, Brenda E; Auyeung, Kate; Bhattacharya, Jayanta; Vajpayee, Madhu; Wig, Naveet; Pantophlet, Ralph; Luthra, Kalpana

    2015-01-01

    Dissecting antibody specificities in the plasma of HIV-1 infected individuals that develop broadly neutralizing antibodies (bNAbs) is likely to provide useful information for refining target epitopes for vaccine design. Several studies have reported CD4-binding site (CD4bs) antibodies as neutralization determinants in the plasma of subtype B-infected individuals; however there is little information on the prevalence of CD4bs specificities in HIV-infected individuals in India. Here, we report on the presence of CD4bs antibodies and their contribution to virus neutralization in the plasma from a cohort of HIV-1 infected Indian individuals. Plasma from 11 of the 140 HIV-1 infected individuals (7.9%) studied here exhibited cross-neutralization activity against a panel of subtype B and C viruses. Analyses of these 11 plasma samples for the presence of CD4bs antibodies using two CD4bs-selective probes (antigenically resurfaced HXB2gp120 core protein RSC3 and hyperglycosylated JRFLgp120 mutant ΔN2mCHO) revealed that five (AIIMS 617, 619, 627, 642, 660) contained RSC3-reactive plasma antibodies and only one (AIIMS 660) contained ΔN2mCHO-reactive antibodies. Plasma antibody depletion and competition experiments confirmed that the neutralizing activity in the AIIMS 660 plasma was dependent on CD4bs antibodies. To the best of our knowledge, this is the first study to report specifically on the presence of CD4bs antibodies in the plasma of a cohort of HIV-1 infected Indian donors. The identification of CD4bs dependent neutralizing antibodies in an HIV-1 infected Indian donor is a salient finding of this study and is supportive of ongoing efforts to induce similar antibodies by immunization.

  19. Identification of CD4-Binding Site Dependent Plasma Neutralizing Antibodies in an HIV-1 Infected Indian Individual.

    Directory of Open Access Journals (Sweden)

    Lubina Khan

    Full Text Available Dissecting antibody specificities in the plasma of HIV-1 infected individuals that develop broadly neutralizing antibodies (bNAbs is likely to provide useful information for refining target epitopes for vaccine design. Several studies have reported CD4-binding site (CD4bs antibodies as neutralization determinants in the plasma of subtype B-infected individuals; however there is little information on the prevalence of CD4bs specificities in HIV-infected individuals in India. Here, we report on the presence of CD4bs antibodies and their contribution to virus neutralization in the plasma from a cohort of HIV-1 infected Indian individuals. Plasma from 11 of the 140 HIV-1 infected individuals (7.9% studied here exhibited cross-neutralization activity against a panel of subtype B and C viruses. Analyses of these 11 plasma samples for the presence of CD4bs antibodies using two CD4bs-selective probes (antigenically resurfaced HXB2gp120 core protein RSC3 and hyperglycosylated JRFLgp120 mutant ΔN2mCHO revealed that five (AIIMS 617, 619, 627, 642, 660 contained RSC3-reactive plasma antibodies and only one (AIIMS 660 contained ΔN2mCHO-reactive antibodies. Plasma antibody depletion and competition experiments confirmed that the neutralizing activity in the AIIMS 660 plasma was dependent on CD4bs antibodies. To the best of our knowledge, this is the first study to report specifically on the presence of CD4bs antibodies in the plasma of a cohort of HIV-1 infected Indian donors. The identification of CD4bs dependent neutralizing antibodies in an HIV-1 infected Indian donor is a salient finding of this study and is supportive of ongoing efforts to induce similar antibodies by immunization.

  20. Endogenous plasma estradiol in healthy men is positively correlated with cerebral cortical serotonin 2A receptor binding

    DEFF Research Database (Denmark)

    Frokjaer, Vibe G.; Erritzoe, David; Juul, Anders;

    2010-01-01

    Background: Sex-hormones influence brain function and are likely to play a role in the gender predisposition to mood and anxiety disorders. Acute fluctuations of sex-hormone levels including hormonal replacement therapy appear to affect serotonergic neurotransmission, but it is unknown if baseline...... = 0.0001), whereas no independent effects of testosterone could be demonstrated. Correction for other factors of importance for 5-HT2A receptor binding did not change the result. A voxel-based analysis suggested that there were no regional differences in the estradiol effect on cortical 5-HT2A...... receptor binding. Conclusions: Our data show a positive correlation between endogenous plasma estradiol levels and cortical 5-HT2A receptor binding in healthy men, whereas, no independent effect of testosterone was demonstrated. We speculate that this association could be mediated through effects on gene...

  1. Technical note: Protozoa-specific antibodies raised in sheep plasma bind to their target protozoa in the rumen.

    Science.gov (United States)

    Williams, Y J; Rea, S M; Popovski, S; Skillman, L C; Wright, A-D G

    2014-12-01

    Binding of IgG antibodies to Entodinium spp. in the rumen of sheep (Ovis aries) was investigated by adding IgG, purified from plasma, directly into the rumen. Plasma IgG was sourced from sheep that had or had not been immunized with a vaccine containing whole fixed Entodinium spp. cells. Ruminal fluid was sampled approximately 2 h after each antibody dosing. Binding of protozoa by a specific antibody was detected using an indirect fluorescent antibody test. An antibody titer in the ruminal fluid was determined by ELISA, and the concentration of ruminal fluid ammonia-N and ruminal pH were also determined. Entodinium spp. and total protozoa from IgG-infused sheep were enumerated by microscopic counts. Two-hourly additions of IgG maintained a low antibody titer in the rumen for 12 h and the binding of the antibody to the rumen protozoa was demonstrated. Increased ammonia-N concentrations and altered ruminal fluid pH patterns indicated that additional fermentation of protein was occurring in the rumen after addition of IgG. No reduction in numbers of Entodinium spp. was observed (P>0.05). Although binding of antibodies to protozoa has been demonstrated in the rumen, it is unclear how much cell death occurred. On the balance of probability, it would appear that the antibody was degraded or partially degraded, and the impact of this on protozoal populations and the measurement of a specific titer is also unclear.

  2. Drug-drug interactions related to altered absorption and plasma protein binding: theoretical and regulatory considerations, and an industry perspective.

    Science.gov (United States)

    Hochman, Jerome; Tang, Cuyue; Prueksaritanont, Thomayant

    2015-03-01

    Drug-drug interactions (DDIs) related to altered drug absorption and plasma protein binding have received much less attention from regulatory agencies relative to DDIs mediated via drug metabolizing enzymes and transporters. In this review, a number of theoretical bases and regulatory framework are presented for these DDI aspects. Also presented is an industry perspective on how to approach these issues in support of drug development. Overall, with the exception of highly permeable and highly soluble (BCS 1) drugs, DDIs related to drug-induced changes in gastrointestinal (GI) physiology can be substantial, thus warranting more attentions. For a better understanding of absorption-associated DDI potential in a clinical setting, mechanistic studies should be conducted based on holistic integration of the pharmaceutical profiles (e.g., pH-dependent solubility) and pharmacological properties (e.g., GI physiology and therapeutic margin) of drug candidates. Although majority of DDI events related to altered plasma protein binding are not expected to be of clinical significance, exceptions exist for a subset of compounds with certain pharmacokinetic and pharmacological properties. Knowledge of the identity of binding proteins and the binding extent in various clinical setting (including disease states) can be valuable in aiding clinical DDI data interpretations, and ensuring safe and effective use of new drugs.

  3. An attempt to validate serum and plasma as sample matrices for analyses of polychlorobiphenylols

    Energy Technology Data Exchange (ETDEWEB)

    Weiss, J.; Bergman, Aa. [Stockholm Univ. (Sweden). Dept. of Environmental Chemistry; Bignert, A. [Museum of Natural History (Sweden)

    2004-09-15

    Polychlorinated biphenyls (PCBs) form hydroxylated metabolites (OH-PCBs), as reported both from wildlife and from experimental animal studies already in the early 1970s'. However, the interest increased in OH-PCBs from the mid 1990s' depending on the discovery that some OHPCB congeners are strongly retained in the blood of birds, fish and mammals, including humans. The interest is linked to the fact that OH-PCBs is strongly, but reversibly, bound to the blood protein transthyretin (TTR). It is reasonable to believe that the strong TTR binding may have toxicological impact, probably related to endocrine type effects. Importantly, OH-PCBs are present in blood at far higher concentrations than in any other compartment in the body, which is dependent on the physico-chemical characteristics of the phenols. Analyses of OH-PCBs have thus been concentrated to whole blood, plasma or serum. Still there is no comparison between the three sample types even though it is clear that whole blood is not optimal due to the large proportion of haemoglobin in the sample that make the clean up more difficult than if plasma or serum is selected for analysis. In the present study we have addressed two questions: First we have looked at any potential differences in the analytical results of OH-PCBs when using serum and plasma for extraction and clean up; Second, the serum and plasma applied in the validation has been unfrozen, frozen (at -20 C) for two months and frozen for twenty months, respectively.

  4. Trojan Tour and Rendezvous (TTR): A New Frontiers Mission to Explore the Origin and Evolution of the Early Solar System

    Science.gov (United States)

    Bell, J. F., III; Olkin, C.; Castillo, J. C.

    2015-12-01

    The orbital properties, compositions, and physical properties of the diverse populations of small outer solar system bodies provide a forensic map of how our solar system formed and evolved. Perhaps the most potentially diagnostic, but least explored, of those populations are the Jupiter Trojan asteroids, which orbit at ~5 AU in the L4 and L5 Lagrange points of Jupiter. More than 6200 Jupiter Trojans are presently known, but these are predicted to be only a small fraction of the 500,000 to 1 million Trojans >1 km in size. The Trojans are hypothesized to be either former Kuiper Belt Objects (KBOs) that were scattered into the inner solar system by early giant planet migration and then trapped in the 1:1 Jupiter mean motion resonance, or bodies formed near 5 AU in a much more quiescent early solar system, and then trapped at L4 and L5. The 2011 Planetary Science Decadal Survey identified important questions about the origin and evolution of the solar system that can be addressed by studying of the Trojan asteroids, including: (a) How did the giant planets and their satellite systems accrete, and is there evidence that they migrated to new orbital positions? (b) What is the relationship between large and small KBOs? Is the small population derived by impact disruption of the large one? (c) What kinds of surface evolution, radiation chemistry, and surface-atmosphere interactions occur on distant icy primitive bodies? And (d) What are the sources of asteroid groups (Trojans and Centaurs) that remain to be explored by spacecraft? The Trojan Tour and Rendezvous (TTR) is a New Frontiers-class mission designed to answer these questions, and to test hypotheses for early giant planet migration and solar system evolution. Via close flybys of a large number of these objects,, and orbital characterization of at least one large Trojan, TTR will enable the first-time exploration of this population. Our primary mission goals are to characterize the overall surface geology

  5. A competition assay to identify amyloidogenesis inhibitors by monitoring the fluorescence emitted by the covalent attachment of a stilbene derivative to transthyretin.

    Science.gov (United States)

    Choi, Sungwook; Kelly, Jeffery W

    2011-02-15

    Herein we demonstrate that competition between candidate kinetic stabilizer binding to transthyretin (TTR) and stilbene binding to and reaction with the same thyroxine sites within TTR can be utilized to discover potent and highly selective non-covalent TTR amyloidogenesis inhibitors. We report two stilbenes, S1 and S2, for use in distinct competition assays. Each bind selectively to TTR and then chemoselectively react to form an amide bond with the Lys-15 residue of TTR, creating a fluorescent conjugate. We used 28 TTR kinetic stabilizers exhibiting a known spectrum of plasma TTR binding selectivities and TTR amyloid fibril inhibition efficacies to validate the 'TTR fluorescence conjugate competition assay'. The kinetic stabilizers competed with S1 for binding to recombinant TTR in buffer and with S2 for binding to endogenous levels of TTR in human blood serum. In both assay scenarios, we demonstrate that the lower the TTR-stilbene conjugate fluorescence after a 3 h competition, the greater the binding selectivity and potency of the candidate TTR kinetic stabilizer. These assays, particularly the assay utilizing S2 in human serum, replace two assays previously utilized to gather the same information. While not the focus of this manuscript, it is clear that the 'TTR fluorescence conjugate competition assay' could be adapted for high throughput screening applications.

  6. Study on the interaction of plasma protein binding rate between edaravone and taurine in human plasma based on HPLC analysis coupled with ultrafiltration technique.

    Science.gov (United States)

    Tang, Dao-quan; Li, Yin-jie; Li, Zheng; Bian, Ting-ting; Chen, Kai; Zheng, Xiao-xiao; Yu, Yan-yan; Jiang, Shui-shi

    2015-08-01

    In this work, two high-performance liquid chromatography (HPLC) assays were developed and validated for the independent determination of edaravone and taurine using 3-methyl-1-p-tolyl-5-pyrazolone and L-glutamine as internal standards. In in vitro experiments, human plasma was separately spiked with a mixture of edaravone and taurine, edaravone or taurine alone. Plasma was precipitated with acetonitrile containing 0.1% formic acid. Ultrafiltration was employed to obtain the unbound ingredients of the two drugs. The factors that might influence the ultrafiltration effiency were elaborately optimized. Plasma supernatant and ultrafiltrate containing taurine were derivated with o-phthalaldehyde and ethanethiol in the presence of 40 mmol/L sodium borate buffer (pH 10.2) at room temperature within 1 min. Chromatographic separations were achieved on an InertSustain C18 column (250 × 4.6 mm, 5 µm). Isocratic 50 mmol/L ammonium acetate-acetonitrile and gradient 50 mmol/L sodium acetate (pH 5.3)-methanol were respectively selected as the mobile phase for the determination of edaravone and taurine. All of the validation data including linearity, extraction recovery, precision, accuracy and stability conformed to the requirements. Results showed that there were no significant alterations in the plasma protein binding rate of taurine and edaravone, implying that the proposed combination therapy was pharmacologically feasible. Copyright © 2014 John Wiley & Sons, Ltd.

  7. Hereditary increase of plasma histidine-rich glycoprotein associated with abnormal heparin binding (HRG Eindhoven).

    Science.gov (United States)

    Hoffmann, J J; Hennis, B C; Kluft, C; Vijgen, M

    1993-12-20

    Plasma histidine-rich glycoprotein (HRG) was found to be persistently increased in a patient with a history of recurrent arterial thromboembolic events. The mean concentration was 270% of normal pooled plasma. Increased HRG was found in eight of the 17 relatives studied, but none of them has experienced thrombo-embolism yet. Apparently, increased HRG was hereditary with autosomal dominant inheritance. A significant correlation was found between the increased plasma concentration of the protein and the age of the subjects (P Eindhoven.

  8. Pi binding by the F1-ATPase of beef heart mitochondria and of the Escherichia coli plasma membrane.

    Science.gov (United States)

    Penefsky, Harvey S

    2005-04-11

    Pi binding by the F(1)-ATPase of beef heart mitochondria and of the Escherichia coli plasma membrane (E. coli F(1)) was examined by two methods: the centrifuge column procedure [Penefsky, H.S. (1977) J. Biol. Chem. 252, 2891-2899] and the Paulus pressure dialysis cell [Paulus, H. (1969) Anal. Biochem. 32, 91-100]. The latter is an equilibrium dialysis-type procedure. Pi binding by beef heart F(1) could be determined by either procedure. However, direct binding of Pi to E. coli F(1) could be determined adequately only in the Paulus cell which indicated more than two binding sites per mol of enzyme with a K(d) in the range of 0.1 mM. It is concluded that previous failure to observe Pi binding to E. coli F(1) with the centrifuge column procedure is due to a rapid rate of dissociation of Pi from the E. coli enzyme which results in loss of Pi during transit of the enzyme-Pi complex through the column.

  9. Identification of calcium-binding proteins associated with the human sperm plasma membrane

    National Research Council Canada - National Science Library

    Naaby-Hansen, Soren; Diekman, Alan; Shetty, Jagathpala; Flickinger, Charles J; Westbrook, Anne; Herr, John C

    2010-01-01

    The precise composition of the human sperm plasma membrane, the molecular interactions that define domain specific functions, and the regulation of membrane associated proteins during the capacitation...

  10. Analysis of the induction of the myelin basic protein binding to the plasma membrane phospholipid monolayer

    Science.gov (United States)

    Zhang, Lei; Hao, Changchun; Feng, Ying; Gao, Feng; Lu, Xiaolong; Li, Junhua; Sun, Runguang

    2016-09-01

    Myelin basic protein (MBP) is an essential structure involved in the generation of central nervous system (CNS) myelin. Myelin shape has been described as liquid crystal structure of biological membrane. The interactions of MBP with monolayers of different lipid compositions are responsible for the multi-lamellar structure and stability of myelin. In this paper, we have designed MBP-incorporated model lipid monolayers and studied the phase behavior of MBP adsorbed on the plasma membrane at the air/water interface by thermodynamic method and atomic force microscopy (AFM). By analyzing the pressure-area (π-A) and pressure-time (π-T) isotherms, univariate linear regression equation was obtained. In addition, the elastic modulus, surface pressure increase, maximal insertion pressure, and synergy factor of monolayers were detected. These parameters can be used to modulate the monolayers binding of protein, and the results show that MBP has the strongest affinity for 1,2-dipalmitoyl-sn-glycero-3- phosphoserine (DPPS) monolayer, followed by DPPC/DPPS mixed and 1,2-dipalmitoyl-sn-glycero-3-phospho-choline (DPPC) monolayers via electrostatic and hydrophobic interactions. AFM images of DPPS and DPPC/DPPS mixed monolayers in the presence of MBP (5 nM) show a phase separation texture at the surface pressure of 20 mN/m and the incorporation of MBP put into the DPPC monolayers has exerted a significant effect on the domain structure. MBP is not an integral membrane protein but, due to its positive charge, interacts with the lipid head groups and stabilizes the membranes. The interaction between MBP and phospholipid membrane to determine the nervous system of the disease has a good biophysical significance and medical value. Project supported by the National Natural Science Foundation of China (Grant Nos. 21402114 and 11544009), the Natural Science Basic Research Plan in Shaanxi Province of China (Grant No. 2016JM2010), the Fundamental Research Funds for the Central

  11. ATP-binding cassette-like transporters are involved in the transport of lignin precursors across plasma and vacuolar membranes

    Energy Technology Data Exchange (ETDEWEB)

    Miao, Y.C.; Liu, C.

    2010-12-28

    Lignin is a complex biopolymer derived primarily from the condensation of three monomeric precursors, the monolignols. The synthesis of monolignols occurs in the cytoplasm. To reach the cell wall where they are oxidized and polymerized, they must be transported across the cell membrane. However, the molecular mechanisms underlying the transport process are unclear. There are conflicting views about whether the transport of these precursors occurs by passive diffusion or is an energized active process; further, we know little about what chemical forms are required. Using isolated plasma and vacuolar membrane vesicles prepared from Arabidopsis, together with applying different transporter inhibitors in the assays, we examined the uptake of monolignols and their derivatives by these native membrane vesicles. We demonstrate that the transport of lignin precursors across plasmalemma and their sequestration into vacuoles are ATP-dependent primary-transport processes, involving ATP-binding cassette-like transporters. Moreover, we show that both plasma and vacuolar membrane vesicles selectively transport different forms of lignin precursors. In the presence of ATP, the inverted plasma membrane vesicles preferentially take up monolignol aglycones, whereas the vacuolar vesicles are more specific for glucoconjugates, suggesting that the different ATP-binding cassette-like transporters recognize different chemical forms in conveying them to distinct sites, and that glucosylation of monolignols is necessary for their vacuolar storage but not required for direct transport into the cell wall in Arabidopsis.

  12. IN SILICO EVALUATION OF ANGIOTENSIN II RECEPTOR ANTAGONIST’S PLASMA PROTEIN BINDING USING COMPUTED MOLECULAR DESCRIPTORS

    Directory of Open Access Journals (Sweden)

    Jadranka Odović

    2014-03-01

    Full Text Available The discovery of new pharmacologically active substances and drugs modeling led to necessity of predicting drugs properties and its ADME data. Angiotensin II receptor antagonists are a group of pharmaceuticals which modulate the renin-angiotensin-aldosterone system and today represent the most commonly prescribed anti-hypertensive drugs. The aim of this study was to compare different molecular properties of seven angiotensin II receptor antagonists / blockers (ARBs, (eprosartan, irbesartan, losartan, olmesartan, telmisartan, valsartan and their plasma protein binding (PPB data. Several ARBs molecular descriptors were calculated using software package Molinspiration Depiction Software as well as Virtual Computational Chemistry Laboratory (electronic descriptor – PSA, constitutional parameter – Mw, geometric descriptor – Vol, lipophilicity descriptors - logP values, aqueous solubility data – logS. The correlations between all collected descriptors and plasma protein binding data obtained from relevant literature were established. In the simple linear regression poor correlations were obtained in relationships between PPB data and all calculated molecular descriptors. In the next stage of the study multiple linear regression (MLR was used for correlation of PPB data with two different descriptors as independent variables. The best correlation (R2=0.70 with P<0.05 was established between PPB data and molecular weight with addition of volume values as independent variables. The possible application of computed molecular descriptors in drugs protein binding evaluation can be of great importance in drug research.

  13. Binding of human myeloperoxidase to red blood cells: Molecular targets and biophysical consequences at the plasma membrane level.

    Science.gov (United States)

    Gorudko, Irina V; Sokolov, Alexey V; Shamova, Ekaterina V; Grigorieva, Daria V; Mironova, Elena V; Kudryavtsev, Igor V; Gusev, Sergey A; Gusev, Alexander A; Chekanov, Andrey V; Vasilyev, Vadim B; Cherenkevich, Sergey N; Panasenko, Oleg M; Timoshenko, Alexander V

    2016-02-01

    Myeloperoxidase (MPO) is an oxidant-producing enzyme that can also bind to cellular surface proteins. We found that band 3 protein and glycophorins A and B were the key MPO-binding targets of human red blood cells (RBCs). The interaction of MPO with RBC proteins was mostly electrostatic in nature because it was inhibited by desialation, exogenic sialic acid, high ionic strength, and extreme pH. In addition, MPO failed to interfere with the lectin-induced agglutination of RBCs, suggesting a minor role of glycan-recognizing mechanisms in MPO binding. Multiple biophysical properties of RBCs were altered in the presence of native (i.e., not hypochlorous acid-damaged) MPO. These changes included transmembrane potential, availability of intracellular Ca(2+), and lipid organization in the plasma membrane. MPO-treated erythrocytes became larger in size, structurally more rigid, and hypersensitive to acidic and osmotic hemolysis. Furthermore, we found a significant correlation between the plasma MPO concentration and RBC rigidity index in type-2 diabetes patients with coronary heart disease. These findings suggest that MPO functions as a mediator of novel regulatory mechanism in microcirculation, indicating the influence of MPO-induced abnormalities on RBC deformability under pathological stress conditions.

  14. High affinity RGD-binding sites at the plasma membrane of Arabidopsis thaliana links the cell wall.

    Science.gov (United States)

    Canut, H; Carrasco, A; Galaud, J P; Cassan, C; Bouyssou, H; Vita, N; Ferrara, P; Pont-Lezica, R

    1998-10-01

    The heptapeptide Tyr-Gly-Arg-Gly-Asp-Ser-Pro containing the sequence Arg-Gly-Asp (RGD--the essential structure recognised by animal cells in substrate adhesion molecules) was tested on epidermal cells of onion and cultured cells of Arabidopsis upon plasmolysis. Dramatic changes were observed on both types of cells following treatment: on onion cells, Hechtian strands linking the cell wall to the membrane were lost, while Arabidopsis cells changed from concave to convex plasmolysis. A control heptapeptide Tyr-Gly-Asp-Gly-Arg-Ser-Pro had no effect on the shape of plasmolysed cells. Protoplasts isolated from Arabidopsis cells agglutinate in the presence of ProNectinF, a genetically engineered protein of 72 kDa containing 13 RGD sequences: several protoplasts may adhere to a single molecule of ProNectinF. The addition of the RGD-heptapeptide disrupted the adhesion between the protoplasts. Purified plasma membrane from Arabidopsis cells exhibits specific binding sites for the iodinated RGD-heptapeptide. The binding is saturable, reversible, and two types of high affinity sites (Kd1 approximately 1 nM, and Kd2 approximately 40 nM) can be discerned. Competitive inhibition by several structurally related peptides and proteins noted the specific requirement for the RGD sequence. Thus, the RGD-binding activity of Arabidopsis fulfils the adhesion features of integrins, i.e. peptide specificity, subcellular location, and involvement in plasma membrane-cell wall attachments.

  15. Low prevalence of antibodies and other plasma factors binding to CC chemokines and IL-2 in HIV-positive patients.

    Science.gov (United States)

    Meyer, C N; Svenson, M; Schade Larsen, C; Odum, N; Skinhøj, P; Bendtzen, K

    2000-02-01

    Neutralizing cytokine antibodies are found in healthy and diseased individuals, including patients treated with recombinant cytokines. Identification of CCR-5 as co-receptor for HIV has focused interest on CC chemokines and their potential therapeutic use. Chemokine-binding components in plasma of HIV-infected patients were therefore assessed by radioimmunoassay and radioreceptor assay. IgG from 4/505 HIV patients and 9/2000 healthy controls (p>0.05) bound rMIP-1alpha and rMIP-1beta, but not rRANTES. No other plasma factors bound the chemokines. The antibodies inhibited receptor binding of both chemokines. There was no association between presence of antibodies and disease stage or HIV progression rate. Three of 11 patients treated with rIL-2 developed IgG antibodies suppressing cellular binding and growth promotion of rIL-2. Hence, circulating factors, including antibodies MIP-1alpha/MIP-1beta, are uncommon in healthy individuals and HIV patients, and are apparently without prognostic significance. In contrast to earlier reports, IL-2 antibodies were found only in HIV patients treated with rIL-2.

  16. Second-generation nanofiltered plasma-derived mannan-binding lectin product

    DEFF Research Database (Denmark)

    Laursen, I.; Houen, G.; Højrup, P.

    2007-01-01

    BACKGROUND AND OBJECTIVES: Mannan-binding lectin (MBL) is an important component of the innate immune defence; it binds to carbohydrate structures on pathogenic micro-organisms resulting in complement activation and opsonization. Individuals with low MBL levels are at risk of recurrent and severe...

  17. Study of plasma protein binding activity of isometamidium and its impact on anthelmintic activity using trypanosoma induced calf model

    Directory of Open Access Journals (Sweden)

    Suprita Sinha

    Full Text Available Aim: The objective of present study was to determine Plasma Protein Binding (PPB activity and its effect on clinical efficacy of isometamidium after intramuscular administration in calves. The binding of drugs to plasma proteins is an important factor in controlling the availability and distribution of drugs. In general, PPB reduces the free fraction of drug available for therapeutic activity, since only the non-protein bound drug is pharmacologically active. Materials and Methods: Six calves were used for PPB study and eighteen for clinical efficacy. Isometamidium was administered @ 0.5mg/kg intramuscularly as a single dose for PPB study. Equilibrium dialysis technique was used to determine the PPB activity. For clinical efficacy, infection with Trypanosoma was induced in calves of two groups, untreated control and experimental group. Infection was confirmed after 28 days by mice inoculation test. Isometamidium @ 0.5mg/kg was administered to experimental group. Haematoobiochemical and mice inoculation tests were performed after 7 days of drug administration (Day 35. Result: The percentage of PPB activity of isometamidium was 86.71 ± 0.59 to 93.03 ± 0.63% against the concentration 9.76± 0.84 to 4.39 ± 0.20 g ml-1. Higher percentage of PPB activity (>86% suggests greater duration of safety by this drug. It was found that anthelmintic activity of isometamidium was substantially affected by higher PPB. Conclusion: It was concluded that isometamidium has greater plasma protein binding capacity which did not hamper clinical efficacy of drug. [Vet World 2013; 6(7.000: 444-448

  18. Characterization of Plasma-Polymerized Fused Polycyclic Compounds for Binding Conducting Polymers

    DEFF Research Database (Denmark)

    Winther-Jensen, Bjørn; Norrman, Kion; Kingshott, Peter

    2005-01-01

    An investigation is made of the plasma polymerization of fused polycyclic monomers containing a dioxy-ring that is fused to an aromatic ring. These molecules provide the basis for very efficient polymerization mechanisms in which only the dioxy-ring undergoes ring opening during the polymerization...... with the remaining part of the monomer remaining intact. XPS, ToF-SIMS, and IR are used to investigate the chemistry of the films produced by plasma polymerization of EDT, which contains a high content of the aromatic group. We find that the plasma-polymerized films of EDT contain intact thiophene groups...

  19. Low prevalence of antibodies and other plasma factors binding to CC chemokines and IL-2 in HIV-positive patients

    DEFF Research Database (Denmark)

    Meyer, C N; Svenson, M; Schade Larsen, C;

    2000-01-01

    of HIV-infected patients were therefore assessed by radioimmunoassay and radioreceptor assay. IgG from 4/505 HIV patients and 9/2000 healthy controls (p>0.05) bound rMIP-1alpha and rMIP-1beta, but not rRANTES. No other plasma factors bound the chemokines. The antibodies inhibited receptor binding of both...... chemokines. There was no association between presence of antibodies and disease stage or HIV progression rate. Three of 11 patients treated with rIL-2 developed IgG antibodies suppressing cellular binding and growth promotion of rIL-2. Hence, circulating factors, including antibodies MIP-1alpha/MIP-1beta...

  20. Simultaneous determination of human plasma protein binding of bioactive flavonoids in Polygonum orientale by equilibrium dialysis combined with UPLC-MS/MS

    Institute of Scientific and Technical Information of China (English)

    Yong Huang; Yong-Lin Wang; Hui Chen; Feng He; Zhi-Rong Zhang; Lin Zheng; Yue Liu; Yan-Yu Lan; Shang-Gao Liao; Yong-Jun Li

    2013-01-01

    A simple and selective ultra performance liquid chromatography-electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS) assay was developed for the determination of the human plasma protein binding of four bioactive flavonoids (such as orientin and vitexin) in Polygonum orientale. Protein precipitation was used for sample preparation. Equilibrium dialysis technique was applied to determine the plasma protein binding under physiological conditions. The separation was achieved through a Waters C18 column with a mobile phase composed of 0.1%formic acid in acetonitrile and 0.1%aqueous formic acid using step gradient elution at a flow rate of 0.35 mL/min. A Waters ACQUITY™TQD system was operated under the multiple reaction monitoring (MRM) mode of positive electrospray ionization. All of the recovery, precision, accuracy and stability of the method met the requirements. Good correlations (r40.99) of the four compounds were found, which suggested that these compounds can be simultaneously determined with acceptable accuracy. Results showed that the plasma protein bindings of the four bioactive flavonoids were in the range of 74-89% over the six concentrations studied. The binding parameters containing protein binding affinity, protein binding dissociation constant, and protein binding site were studied. The maximum ability to bind with protein was also determined in the assay in order to understand the drug-protein binding of each compound better.

  1. Interaction between Fibrinogen and Insulin-Like Growth Factor-Binding Protein-1 in Human Plasma under Physiological Conditions.

    Science.gov (United States)

    Gligorijević, N; Nedić, O

    2016-02-01

    Fibrinogen is a plasma glycoprotein and one of the principle participants in blood coagulation. It interacts with many proteins during formation of a blood clot, including insulin-like growth factors (IGFs) and their binding proteins (IGFBP). Fibrinogen complexes were found as minor fractions in fibrinogen preparations independently of the coagulation process, and their presence influences the kinetics of polymerization. The idea of this work was to investigate whether fibrinogen in human plasma interacts with IGFBPs independently of the tissue injury or coagulation process. The results have shown that fibrinogen forms complexes with IGFBP-1 under physiological conditions. Several experimental approaches have confirmed that complexes are co-isolated with fibrinogen from plasma, they are relatively stable, and they appear as a general feature of human plasma. Several other experiments excluded the possibility that alpha-2 macroglobulin/IGFBP-1 complexes or IGFBP-1 oligomers contributed to IGFBP-1 immunoreactivity. The role of fibrinogen/IGFBP-1 complexes is still unknown. Further investigation in individuals expressing both impaired glucose control and coagulopathy could contribute to identification and understanding of their possible physiological role.

  2. Performance Assessment of Hazardous Air Pollutant (HAP)Free Chemical Paint Strippers on Military Coatings for Validation to Federal Specification TT-R-2918A

    Science.gov (United States)

    2016-03-01

    Free Chemical Paint Strippers on Military Coatings for Validation to Federal Specification TT-R-2918A by Lindsey Blohm Oak Ridge Institute for... chemical -agent- resistant coating (CARC) painted panels is shown in Fig. 1. The top layers of topcoat are tan and the bottom layers of topcoat are green...methylene-chloride-based strippers for removing Army chemical agent resistant coatings (CARC). Aberdeen Proving Ground (MD): Army Research

  3. Measurement of intact insulin-like growth factor-binding protein-3 in human plasma using a ligand immunofunctional assay.

    Science.gov (United States)

    Lassarre, C; Binoux, M

    2001-03-01

    Limited proteolysis of insulin-like growth factor binding protein-3 (IGFBP-3) is a fundamental mechanism in the regulation of IGF-I bioavailability in the bloodstream. Its measurement by Western immunoblotting provides only semiquantitative estimation. We have developed a ligand immunofunctional assay (LIFA) for quantifying human (h) intact IGFBP-3 in biological fluids. IGFBP-bound IGFs are dissociated and separated by acid pH ultrafiltration, and a monoclonal antibody specific to the first 160 amino acids of IGFBP-3 is used to capture hIGFBP-3 in a solid-phase assay. The complex is then incubated with (125)I-IGF-I, which binds to intact IGFBP-3 but not to its proteolytic fragments. Binding specificity was demonstrated in competition experiments with unlabeled IGF. Nonspecific binding was 1.4%. The fragments comprising residues 1-160 and 1-95 of recombinant hIGFBP-3 [corresponding to the major proteolytic fragments of approximately 30 kDa and (glycosylated) 20 or (nonglycosylated) 16 kDa detected in serum by Western immunoblotting, respectively] fail to bind (125)I-IGF-I when complexed with the monoclonal antibody. Similarly, no binding of (125)I-IGF-I was obtained in the LIFA when applied to plasmas from pregnant women during the final 3 months of pregnancy, where the characteristic 42- to 39-kDa doublet of intact IGFBP-3 is undetectable. The standard curve was established using a pool of plasmas (EDTA) from healthy adults, for which standardization with glycosylated recombinant hIGFBP-3 yielded an intact IGFBP-3 content of 2 microg/mL. The dynamic range of the LIFA was 0.50-3.75 microL equivalent of the plasma pool in a total volume of 300 microL per assay tube, with a sensitivity threshold of approximately 1 ng intact IGFBP-3. Unknown plasma samples were studied at three concentrations. Intra- and interassay variations were 3.6% and 4%, respectively. In 31 healthy adults, the mean plasma concentration of intact IGFBP-3 was 2.24 +/- 0.08 (SEM) mg/L, and that of

  4. Closure Report for Corrective Action Unit 408: Bomblet Target Area Tonopah Test Range (TTR), Nevada, Revision 0

    Energy Technology Data Exchange (ETDEWEB)

    Mark Krauss

    2010-09-01

    This Closure Report (CR) presents information supporting the closure of Corrective Action Unit (CAU) 408: Bomblet Target Area (TTR), Tonopah Test Range, Nevada. This CR complies with the requirements of the Federal Facility Agreement and Consent Order that was agreed to by the State of Nevada; U.S. Department of Energy (DOE), Environmental Management; U.S. Department of Defense; and DOE, Legacy Management. Corrective Action Unit 408 is located at the Tonopah Test Range, Nevada, and consists of Corrective Action Site (CAS) TA-55-002-TAB2, Bomblet Target Areas. This CAS includes the following seven target areas: • Mid Target • Flightline Bomblet Location • Strategic Air Command (SAC) Target Location 1 • SAC Target Location 2 • South Antelope Lake • Tomahawk Location 1 • Tomahawk Location 2 The purpose of this CR is to provide documentation supporting the completed corrective actions and data confirming that the closure objectives for the CAS within CAU 408 were met. To achieve this, the following actions were performed: • Review the current site conditions, including the concentration and extent of contamination. • Implement any corrective actions necessary to protect human health and the environment. • Properly dispose of corrective action and investigation wastes. • Document Notice of Completion and closure of CAU 408 issued by the Nevada Division of Environmental Protection. From July 2009 through August 2010, closure activities were performed as set forth in the Streamlined Approach for Environmental Restoration Plan for CAU 408: Bomblet Target Area, Tonopah Test Range (TTR), Nevada. The purposes of the activities as defined during the data quality objectives process were as follows: • Identify and remove munitions of explosive concern (MEC) associated with DOE activities. • Investigate potential disposal pit locations. • Remove depleted uranium-contaminated fragments and soil. • Determine whether contaminants of concern (COCs) are

  5. Heparin-binding proteins of seminal plasma in Nellore bulls Proteínas ligadoras à heparina do plasma seminal em touros Nelore

    Directory of Open Access Journals (Sweden)

    Carlos Eurico Fernandes

    2009-02-01

    Full Text Available The aim of this study was to identify heparin-binding proteins (HBPs in seminal plasma of Nellore (Bos taurus indicus bulls. Bulls (n=4, 30-36 months old, 500-550kg with satisfactory seminal quality were selected. After the centrifugation, samples of the seminal plasma were pooled and the HBPs were isolated by heparin-affinity chromatography. The recovered HBPs fractions were pooled. One-dimensional sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDSPAGE 12.5% was performed in vertical minigels. Eight bands with molecular weights ranging from 15 to 63kDa were observed. Two proteins were identified (22 and 25kDa, similar to those previously described in Bos taurus taurus bulls. Other bands identified in this study (39, 53, 58 and 63kDa have not been previously observed and possibly they are specific to Nellore semen.O objetivo deste estudo foi identificar proteínas ligadoras à heparina no plasma seminal de touros Nelore (Bos taurus indicus. Para tanto, foram selecionados quatro touros entre 30 e 36 meses de idade e peso aproximado de 500-550kg. Após centrifugação, amostras do plasma seminal foram misturadas e as proteínas ligadoras à heparina foram isoladas por meio da cromatografia por afinidade. As frações após a eluição foram agrupadas para caracterização das bandas protéicas (SDSPAGE, 12,5%. Foram identificadas oito bandas protéicas variando entre 15 e 63kDa. Duas proteínas com 22 e 25kDa foram similares às descritas em touros Bos taurus taurus. Outras proteínas identificadas com 39, 53, 58 e 63kDa ainda não foram descritas e possivelmente sejam específicas para Bos taurus indicus.

  6. Active seepage and water infiltration in Lake Baikal sediments: new thermal data from TTR-Baikal 2014 (Class@Baikal)

    Science.gov (United States)

    Poort, Jeffrey; Khlystov, Oleg M.; Akhmanov, Grigorii G.; Khabuev, Andrei V.; Belousov, Oleg V.

    2015-04-01

    New thermal data from the sediments of Lake Baikal were collected in July 2014 during the first Training-Through-Research cruise on Lake Baikal (Class@Baikal) organized by MGU and LIN. TTR-Baikal is a comprehensive multidisciplinary program to train students on the field on pertinent scientific topics. The cruise program focused on seafloor sampling, acoustic investigations and heat flow measurements of gas seeps, flares, mud volcanoes, slumps and debris flows, canyons and channels in the coastal proximity. The thermal data were acquired using autonomous temperature sensors on a 3 meter long gravity corer that allowed analysis at the same spot of sediments, pore fluids, hydrates and microbiology. A total of eight thermal measurements were performed in five structures located on the lake floor of the Central Baikal Basin at 333-1530 meter water depths: 3 mud volcanoes (Novosibirsk, Unshuy and Krest), 1 seep site (Seep 13), and one fault outcrop in the Selenga transfer zone. All studied structures show signals of active seepage, water infiltration and/or hydrate dynamics. The strongest thermal gradient has been measured in Seep 13, suggesting a strong upflow of warm fluids similar to the Gorevoy Utes seep. At the three mud volcanoes, hydrate presence have been evidenced and both enhanced and reduced thermal gradients have been observed. This is similar to the hydrate-bearing K-2 mud volcano in Baikal (Poort et al., 2012). A strongly reduced thermal gradient was observed in the Krest mud volcano where the presence of oxidized channels at 30-40 cm under the sediment surface indicate an infiltration of cold lake water. The water infiltration process at hydrate bearing seep sites will be discussed and compared with other seep areas in the world.

  7. Isolation, purification and characterization of beta-1,3-glucan binding protein from the plasma of marine mussel Perna viridis.

    Science.gov (United States)

    Jayaraj, S S; Thiagarajan, R; Arumugam, M; Mullainadhan, P

    2008-06-01

    A beta-1,3-glucan binding protein (betaGBP) specific for laminarin (a beta-1,3-glucan) was detected for the first time in a mollusc, Perna viridis. betaGBP was isolated and purified from the plasma using laminarin precipitation and affinity chromatography on laminarin-Sepharose 6B, respectively. It agglutinated bakers yeast, bacteria, and erythrocytes and enhanced prophenoloxidase (proPO) activity of the plasma in a dose-dependent manner. The purified betaGBP appeared as a single band in native-PAGE and the purity was conformed by HPLC. The protein has a molecular weight estimate of 510kDa as determined by SDS-PAGE and in isoelectric focusing the purified betaGBP was focused as a single band at pI 5.3. betaGBP was found to possess inherent serine protease activity but lacked beta-1,3-glucanase activity and all these results suggest that plasma betaGBP of P. viridis functions as a recognition molecule for beta-1,3-glucan on the surface of microbial cell walls. This recognition and binding lead to the activation of the prophenoloxidase cascade mediated by the inherent serine protease activity of betaGBP. Presence of agglutinating activity and serine protease activity shows that betaGBP is a bifunctional protein. The findings are discussed in light of the importance of this protein in the innate immune response of P. viridis, and they implicate evolutionary link with similar proteins found in other invertebrates.

  8. Vitamin D-Binding Protein Levels in Plasma and Gingival Crevicular Fluid of Patients with Generalized Aggressive Periodontitis

    Directory of Open Access Journals (Sweden)

    Xin Zhang

    2014-01-01

    Full Text Available Vitamin D-binding protein (DBP is the main transport protein of vitamin D and plays an important role in the immune system and host defenses. The purpose of this study was to measure DBP levels in plasma and gingival crevicular fluid (GCF of patients with generalized aggressive periodontitis (GAgP, in comparison to healthy controls, with the goal of elucidating the relationship between DBP and GAgP. Fifty-nine GAgP patients and 58 healthy controls were recruited for the study; clinical parameters of probing depths (PD, bleeding index, and attachment loss (AL were recorded. DBP levels were measured by enzyme-linked immunosorbent assay. From the results, GAgP patients had higher plasma DBP concentrations (P<0.001 but lower GCF DBP concentrations (P<0.001 than healthy controls. In GAgP group, after controlling the potential confounders of age, gender, smoking status, and BMI index, GCF DBP concentrations correlated negatively with PD (P<0.001 and AL (P=0.009. Within the limits of the study, we concluded that decreased GCF DBP level and increased plasma DBP level are associated with periodontitis.

  9. The ATP-binding cassette transporter-2 (ABCA2) regulates esterification of plasma membrane cholesterol by modulation of sphingolipid metabolism.

    Science.gov (United States)

    Davis, Warren

    2014-01-01

    The ATP-binding cassette transporters are a large family (~48 genes divided into seven families A-G) of proteins that utilize the energy of ATP-hydrolysis to pump substrates across lipid bilayers against a concentration gradient. The ABC "A" subfamily is comprised of 13 members and transport sterols, phospholipids and bile acids. ABCA2 is the most abundant ABC transporter in human and rodent brain with highest expression in oligodendrocytes, although it is also expressed in neurons. Several groups have studied a possible connection between ABCA2 and Alzheimer's disease as well as early atherosclerosis. ABCA2 expression levels have been associated with changes in cholesterol and sphingolipid metabolism. In this paper, we hypothesized that ABCA2 expression level may regulate esterification of plasma membrane-derived cholesterol by modulation of sphingolipid metabolism. ABCA2 overexpression in N2a neuroblastoma cells was associated with an altered bilayer distribution of the sphingolipid ceramide that inhibited acylCoA:cholesterol acyltransferase (ACAT) activity and cholesterol esterification. In contrast, depletion of endogenous ABCA2 in the rat schwannoma cell line D6P2T increased esterification of plasma membrane cholesterol following treatment with exogenous bacterial sphingomyelinase. These findings suggest that control of ABCA2 expression level may be a key locus of regulation for esterification of plasma membrane-derived cholesterol through modulation of sphingolipid metabolism.

  10. Study on the plasma protein binding rate of Schisandra lignans based on the LC-IT-TOF/MS technique with relative quantitative analysis.

    Science.gov (United States)

    Liang, Yan; Zhou, Yuan-Yuan; Liu, Yan-Na; Guan, Tian-Ye; Zheng, Xiao; Dai, Chen; Xing, Lu; Rao, Tai; Xie, Lin; Wang, Guang-Ji

    2013-07-01

    The main objective of the current study was to develop a universal method for a protein binding assay of complicated herbal components, and to investigate the possible relationship between compound polarity and protein binding using Schisadra lignans as an example. Firstly, the rat, dog and human plasma were spiked with three different concentrations of Schisandra chinensis extract (SLE), and ultramicrofiltration was used to obtain the unbound ingredients. Secondly, thirty-one Schisandra lignans in total plasma and ultrafiltered fluid were measured by LC-IT-TOFMS. Lastly, a relative exposure approach, which entailed calculating the relative concentrations of each Schisandra lignan from the corresponding calibration equation created from the calibration samples spiked with the stock solution of SLE, was applied in order to overcome the absence of authentic standards. The results showed that Schisandra lignans exhibited a high capability to bind with plasma protein, furthermore, the protein binding ratio of the lignan components increased proportionally with their individual chromatographic retention time, which indicated that the ratio of protein binding of lignans might increase accordingly with decreasing polarity. This study suggested that the compound polarity might be an important factor affecting the plasma protein binding of herbal components.

  11. Serum and plasma fibronectin binds to complement reacted immune complexes primarily via Clq

    DEFF Research Database (Denmark)

    Baatrup, G; Svehag, S E

    1986-01-01

    The binding of fibronectin to human Clq, C3b, and complement-reacted immune complexes (IC) was investigated by enzyme-linked immunosorbent assays. Microplates were coated with BSA followed by incubation with rabbit-anti-BSA IgG or F(ab')2 fragments of rabbit anti-BSA. Incubation of the solid phas...

  12. Increased Plasma Levels of Heparin-Binding Protein on Admission to Intensive Care Are Associated with Respiratory and Circulatory Failure.

    Directory of Open Access Journals (Sweden)

    Jonas Tydén

    Full Text Available Heparin-binding protein (HBP is released by granulocytes and has been shown to increase vascular permeability in experimental investigations. Increased vascular permeability in the lungs can lead to fluid accumulation in alveoli and respiratory failure. A generalized increase in vascular permeability leads to loss of circulating blood volume and circulatory failure. We hypothesized that plasma concentrations of HBP on admission to the intensive care unit (ICU would be associated with decreased oxygenation or circulatory failure.This is a prospective, observational study in a mixed 8-bed ICU. We investigated concentrations of HBP in plasma at admission to the ICU from 278 patients. Simplified acute physiology score (SAPS 3 was recorded on admission. Sequential organ failure assessment (SOFA scores were recorded daily for three days.Median SAPS 3 was 58.8 (48-70 and 30-day mortality 64/278 (23%. There was an association between high plasma concentrations of HBP on admission with decreased oxygenation (p<0.001 as well as with circulatory failure (p<0.001, after 48-72 hours in the ICU. There was an association between concentrations of HBP on admission and 30-day mortality (p = 0.002. ROC curves showed areas under the curve of 0,62 for decreased oxygenation, 0,65 for circulatory failure and 0,64 for mortality.A high concentration of HBP in plasma on admission to the ICU is associated with respiratory and circulatory failure later during the ICU care period. It is also associated with increased 30-day mortality. Despite being an interesting biomarker for the composite ICU population it's predictive value at the individual patient level is low.

  13. The recombinant LIC10508 is a plasma fibronectin, plasminogen, fibrinogen and C4BP-binding protein of Leptospira interrogans.

    Science.gov (United States)

    Siqueira, Gabriela H; Teixeira, Aline F; Fernandes, Luis G; de Souza, Gisele O; Kirchgatter, Karin; Romero, Eliete C; Vasconcellos, Silvio A; Vieira, Monica L; Nascimento, Ana Lucia T O

    2016-03-01

    Leptospirosis is a zoonosis caused by pathogenic Leptospira spp. In this study, we report that the recombinant proteins LIC10507, LIC10508 and LIC10509 are recognized by confirmed leptospirosis serum samples at both phases of the disease. The recombinant rLIC10508 and rLIC10507 are plasminogen (PLG)-binding proteins, capable of generating plasmin in the presence of a PLG activator. The proteins bind to PLG in a dose-dependent and saturable manner, fulfilling host-ligand interaction. Furthermore, rLIC10508 interacts with fibrinogen (Fg), plasma fibronectin and C4b binding protein (C4BP). The binding of rLIC10508 to Fg decreases the fibrin clotting in a thrombin-catalyzed reaction. The incubation with 4 μM of protein promoted 40% inhibition upon clotting formation. C4BP bound to rLIC10508 retained its cofactor activity for factor I promoting the cleavage of C4b protein, which may reduce the membrane attack complex formation. Although these proteins have high amino acid sequence similarity, rLIC10508 is the most talented of the three, a behavior that might be explained by its unique putative 3D structure, whereas structures of rLIC10507 and rLIC10509 are very similar. Plasmin generation (rLIC10507 and rLIC10508), together with decreasing fibrin clot formation (rLIC10508) and impairment of the complement system (rLIC10508) may help the bacteria to overcome host defense, facilitating the infection process.

  14. Spectrophotometric determination of total proteins in blood plasma: a comparative study among dye-binding methods

    OpenAIRE

    Dimas Augusto Morozin Zaia; Fábio Rangel Marques; Cássia Thaïs Bussamra Vieira Zaia

    2005-01-01

    A comparative study between the biuret method (standard method for total proteins) and spectrophotometric methods using dyes (Bradford, 3',3",5',5"-tetrabromophenolphthalein ethyl ester-TBPEE, and erythrosin-B) was carried out for the determination of total proteins in blood plasma from rats. Bradford method showed the highest sensitivity for proteins and biuret method showed the lowest. For all the methods, the absorbance for different proteins (BSA, casein, and egg albumin) was measured and...

  15. Surface modification by cold-plasma technique for dental implants—Bio-functionalization with binding pharmaceuticals

    Directory of Open Access Journals (Sweden)

    Masao Yoshinari

    2011-08-01

    At the bone tissue/implant interface, a thin calcium phosphate coating and rapid heating with infrared radiation were effective in controlling the dissolution without cracking the coating. These thin calcium phosphate coatings may directly promote osteogenisis, but also enable immobilization and subsequent drug delivery system (DDS of bisphosphonates. Simvastatin is also an effective candidate that is reported to increase the expression of BMP-2. The thin-film of hexamethyldisiloxane (HMDSO was plasma-polymerized onto titanium, and then HMDSO surface was activated by O2-plasma treatment. A quartz crystal microbalance (QCM-D technique demonstrated that simvastatin was immobilized on the plasma-treated surfaces due to introduction of O2-functional groups. At the soft tissue/implant interface, multi-grooved surface topographies and utilizing the adhesive proteins such as fibronectin or laminin-5 may help in providing a biological seal around the implant. At the oral fluid/implant interface, an alumina coating, F+-implantation and immobilization of anti-microbial peptides were responsible for inhibiting the biofilm accumulation.

  16. The concentration of the C-type lectin, mannan-binding protein, in human plasma increases during an acute phase response

    DEFF Research Database (Denmark)

    Thiel, S; Holmskov, U; Hviid, L

    1992-01-01

    Two ELISAs for estimating mannan-binding protein (MBP) were constructed and the concentration of MBP in plasma was followed in patients undergoing major surgery and in patients having a malarial attack. In both cases increases of MBP in the plasma were observed. The relative increase and the kine......Two ELISAs for estimating mannan-binding protein (MBP) were constructed and the concentration of MBP in plasma was followed in patients undergoing major surgery and in patients having a malarial attack. In both cases increases of MBP in the plasma were observed. The relative increase...... and the kinetics varied from person to person. The concentration of MBP increased between 1.5- and three-fold following surgery. In some patients an increase was seen at day 1 whereas in others the increase was not observed until days 3-9. In the malaria patients an increased level of MBP was maintained during 30...

  17. Bioanalysis for plasma protein binding studies in drug discovery and drug development: views and recommendations of the European Bioanalysis Forum.

    Science.gov (United States)

    Buscher, Brigitte; Laakso, Sirpa; Mascher, Hermann; Pusecker, Klaus; Doig, Mira; Dillen, Lieve; Wagner-Redeker, Winfried; Pfeifer, Thomas; Delrat, Pascal; Timmerman, Philip

    2014-03-01

    Plasma protein binding (PPB) is an important parameter for a drug's efficacy and safety that needs to be investigated during each drug-development program. Even though regulatory guidance exists to study the extent of PPB before initiating clinical studies, there are no detailed instructions on how to perform and validate such studies. To explore how PPB studies involving bioanalysis are currently executed in the industry, the European Bioanalysis Forum (EBF) has conducted three surveys among their member companies: PPB studies in drug discovery (Part I); in vitro PPB studies in drug development (Part II); and in vivo PPB studies in drug development. This paper reflects the outcome of the three surveys, which, together with the team discussions, formed the basis of the EBF recommendation. The EBF recommends a tiered approach to the design of PPB studies and the bioanalysis of PPB samples: 'PPB screening' experiments in (early) drug discovery versus qualified/validated procedures in drug development.

  18. Insulin sensitivity is independent of lipid binding protein trafficking at the plasma membrane in human skeletal muscle

    DEFF Research Database (Denmark)

    Jordy, Andreas Børsting; Serup, Annette Karen; Karstoft, Kristian

    2014-01-01

    The aim of the present study was to investigate lipid-induced regulation of lipid binding proteins in human skeletal muscle and the impact hereof on insulin sensitivity. Eleven healthy male subjects underwent a 3-day hyper-caloric and high-fat diet regime. Muscle biopsies were taken before......-regulated by increased fatty acid availability. This suggests a time dependency in the up-regulation of FAT/CD36 and FABPpm protein during high availability of plasma fatty acids. Furthermore, we did not detect FATP1 and FATP4 protein in giant sarcolemmal vesicles obtained from human skeletal muscle. In conclusion......, this study shows that a short-term lipid-load increases mRNA content of key lipid handling proteins in human muscle. However, decreased insulin sensitivity after high-fat diet is not accompanied with relocation of FAT/CD36 or FABPpm protein to the sarcolemma. Finally, FATP1 and FATP4 protein could...

  19. Mechanisms of recognition and binding of α-TTP to the plasma membrane by multi-scale molecular dynamics simulations

    Directory of Open Access Journals (Sweden)

    Christos eLamprakis

    2015-07-01

    Full Text Available We used multiple sets of simulations both at the atomistic and coarse-grained level of resolution, to investigate interaction and binding of α-tochoperol transfer protein (α-TTP to phosphatidylinositol phosphate lipids (PIPs. Our calculations indicate that enrichment of membranes with such lipids facilitate membrane anchoring. Atomistic models suggest that PIP can be incorporated into the binding cavity of α-TTP and therefore confirm that such protein can work as lipid exchanger between the endosome and the plasma membrane. Comparison of the atomistic models of the α-TTP / PIPs complex with membrane-bound α-TTP revealed different roles for the various basic residues composing the basic patch that is key for the protein / ligand interaction. Such residues are of critical importance as several point mutations at their position lead to severe forms of ataxia with vitamin E deficiency (AVED phenotypes. Specifically, R221 is main residue responsible for the stabilisation of the complex. R68 and R192 exchange strong interactions in the protein or in the membrane complex only, suggesting that the two residues alternate contact formation, thus facilitating lipid flipping from the membrane into the protein cavity during the lipid exchange process. Finally, R59 shows weaker interactions with PIPs anyway with a clear preference for specific phosphorylation positions, hinting a role in early membrane selectivity for the protein. Altogether, our simulations reveal significant aspects at the atomistic scale of interactions of α-TTP with the plasma membrane and with PIP, providing clarifications on the mechanism of intracellular vitamin E trafficking and helping establishing the role of key residue for the functionality of α-TTP.

  20. Spectrophotometric determination of total proteins in blood plasma: a comparative study among dye-binding methods

    Directory of Open Access Journals (Sweden)

    Dimas Augusto Morozin Zaia

    2005-05-01

    Full Text Available A comparative study between the biuret method (standard method for total proteins and spectrophotometric methods using dyes (Bradford, 3',3",5',5"-tetrabromophenolphthalein ethyl ester-TBPEE, and erythrosin-B was carried out for the determination of total proteins in blood plasma from rats. Bradford method showed the highest sensitivity for proteins and biuret method showed the lowest. For all the methods, the absorbance for different proteins (BSA, casein, and egg albumin was measured and Bradford method showed the lowest variation of absorbance. The concentration of total protein obtained by using Bradford method was not statistically different (p>0.05 from concentration of total protein obtained by the biuret method. But in regard to erythrosin-B and TBPEE methods the concentrations of total protein were statistically different (pA determinação de proteínas totais em plasma sangüíneo é importante em diversas áreas de pesquisa. Um estudo comparativo entre o método de biureto (método padrão para proteínas totais e diversos métodos que utilizam corantes (Bradford, tetrabromofenolftaleína etil éster-TBPEE, e eritrosina-B foi realizado para a determinação de proteínas totais em plasma sangüíneo de ratos. O método de Bradford mostrou a maior sensibilidade para proteínas e o de biureto a menor. Para todos os métodos, as absorbâncias para diferentes proteínas (BSA, caseína, e ovoalbumina foram medidas e o método de Bradford mostrou a menor variação da absorbância. Utilizando o método de Bradford a concentração de proteínas totais obtida não foi estatisticamente diferente (p>0.05 daquela obtida pelo método do biureto. Porém, para os métodos da eritrosina-B e TBPEE as concentrações de proteínas totais foram estatisticamente diferentes (p<0.05 da obtida pelo método de biureto. Portanto o método de Bradford pode ser utilizado no lugar do método de biureto para a determinação de proteínas totais em plasma sangüíneo.

  1. A strategy for increasing the brain uptake of a radioligand in animals: use of a drug that inhibits plasma protein binding

    Energy Technology Data Exchange (ETDEWEB)

    Haradahira, Terushi E-mail: terushi@nirs.go.jp; Zhang, Ming-Rong; Maeda, Jun; Okauchi, Takashi; Kawabe, Kouichi; Kida, Takayo; Suzuki, Kazutoshi; Suhara, Tetsuya

    2000-05-01

    A positron-emitter labeled radioligand for the glycine-binding site of the N-methyl-D-aspartate (NMDA) receptor, [{sup 11}C]L-703,717, was examined for its ability to penetrate the brain in animals by simultaneous use with drugs having high-affinity separate binding sites on human serum albumin. [{sup 11}C]L-703,717 has poor blood-brain barrier (BBB) permeability because it binds tightly to plasma proteins. Co-injection of warfarin (50-200 mg/kg), a drug that binds to albumin and resembles L-703,717 in structure, dose-dependently enhanced the penetration by [{sup 11}C]L-703,717 in mice, resulting in a five-fold increase in the brain radioactivity at 1 min after the injection. Drugs structurally unrelated to L-703,717, salicylate, phenol red, and L-tryptophan, were less effective or ineffective in increasing the uptake of [{sup 11}C]L-703,717. These results suggest that the simultaneous use of a drug that inhibits the binding of a radioligand to plasma proteins is a useful way to overcome the poor BBB permeability of the radioligand triggered by its tight binding to plasma proteins. In brain distribution studies in rodents, it was found that, after the increase in brain uptake with warfarin, much of the glycine site antagonist accumulates in the cerebellum but its pharmacological specificity did not match the glycine site of NMDA receptors.

  2. A physiologically based pharmacokinetic model to predict the pharmacokinetics of highly protein-bound drugs and the impact of errors in plasma protein binding.

    Science.gov (United States)

    Ye, Min; Nagar, Swati; Korzekwa, Ken

    2016-04-01

    Predicting the pharmacokinetics of highly protein-bound drugs is difficult. Also, since historical plasma protein binding data were often collected using unbuffered plasma, the resulting inaccurate binding data could contribute to incorrect predictions. This study uses a generic physiologically based pharmacokinetic (PBPK) model to predict human plasma concentration-time profiles for 22 highly protein-bound drugs. Tissue distribution was estimated from in vitro drug lipophilicity data, plasma protein binding and the blood: plasma ratio. Clearance was predicted with a well-stirred liver model. Underestimated hepatic clearance for acidic and neutral compounds was corrected by an empirical scaling factor. Predicted values (pharmacokinetic parameters, plasma concentration-time profile) were compared with observed data to evaluate the model accuracy. Of the 22 drugs, less than a 2-fold error was obtained for the terminal elimination half-life (t1/2 , 100% of drugs), peak plasma concentration (Cmax , 100%), area under the plasma concentration-time curve (AUC0-t , 95.4%), clearance (CLh , 95.4%), mean residence time (MRT, 95.4%) and steady state volume (Vss , 90.9%). The impact of fup errors on CLh and Vss prediction was evaluated. Errors in fup resulted in proportional errors in clearance prediction for low-clearance compounds, and in Vss prediction for high-volume neutral drugs. For high-volume basic drugs, errors in fup did not propagate to errors in Vss prediction. This is due to the cancellation of errors in the calculations for tissue partitioning of basic drugs. Overall, plasma profiles were well simulated with the present PBPK model. Copyright © 2016 John Wiley & Sons, Ltd.

  3. Common FABP4 Genetic Variants and Plasma Levels of Fatty Acid Binding Protein 4 in Older Adults

    Science.gov (United States)

    Mukamal, Kenneth J.; Wilk, Jemma B.; Biggs, Mary L.; Jensen, Majken K.; Ix, Joachim H.; Kizer, Jorge R.; Tracy, Russell P.; Zieman, Susan J.; Mozaffarian, Dariush; Psaty, Bruce M.; Siscovick, David S.; Djoussé, Luc

    2013-01-01

    We examined common variants in the fatty acid binding protein 4 gene (FABP4) and plasma levels of FABP4 in adults aged 65 and older from the Cardiovascular Health Study. We genotyped rs16909187, rs1054135, rs16909192, rs10808846, rs7018409, rs2290201, and rs6992708 and measured circulating FABP4 levels among 3190 European Americans and 660 African Americans. Among European Americans, the minor alleles of six single nucleotide polymorphisms (SNP) were associated with lower FABP4 levels (all p ≤ 0.01). Among African Americans, the SNP with the lowest minor allele frequency was associated with lower FABP4 levels (p = 0.015). The C-A haplotype of rs16909192 and rs2290201 was associated with lower FABP4 levels in both European Americans (frequency = 16 %; p = 0.001) and African Americans (frequency = 8 %; p = 0.04). The haplotype combined a SNP in the first intron with one in the 3′untranslated region. However, the alleles associated with lower FABP4 levels were associated with higher fasting glucose in meta-analyses from the MAGIC consortium. These results demonstrate associations of common SNP and haplotypes in the FABP4 gene with lower plasma FABP4 but higher fasting glucose levels. PMID:24043587

  4. Purification and characterization of a native zinc-binding high molecular weight multiprotein complex from human seminal plasma.

    Science.gov (United States)

    Yadav, Vikash Kumar; Kumar, Vijay; Chhikara, Nirmal; Kumar, Sanjay; Manral, Pallavi; Kashav, Tara; Saini, Savita; Srinivasan, A; Singh, Sarman; Singh, Tej P; Yadav, Savita

    2011-05-01

    The seminal plasma comprises secretions from various accessory sex glands. During fertilization spermatozoa undergo complex sequences of precisely timed events that are regulated by the activation of different intracellular signaling pathways. The precision and efficacy of these pathways are often influenced by the assembly and interactions of multiprotein complexes, thereby directing the flow of regulatory information. Our knowledge about these protein complexes present in human seminal plasma (HuSP) is limited. Here we report the identification and characterization of a native high molecular weight zinc-binding multiprotein complex from HuSP by utilizing 2-DE followed by MS. Twenty-six proteins representing isoforms and/or fragments of 11 different proteins were found to be assembled in this complex. Prostate-specific antigen, zinc α2-glycoprotein, prostatic acid phosphatase, and prolactin inducible protein were the major proteins of this complex. Dynamic light scattering experiments revealed changes in aggregation pattern accompanied with deviation from physiological pH and in presence of SDS. However, no significant changes were observed in the presence of physiological ligands such as zinc and fructose. The present study will be useful and contribute to guide the future studies performed for elucidation of biological significance of this native complex in HuSP.

  5. Plasma Free Fatty Acids, Fatty Acid-binding Protein 4, and Mortality in Older Adults (From the Cardiovascular Health Study)

    Science.gov (United States)

    Miedema, Michael D.; Maziarz, Marlena; Biggs, Mary L.; Zieman, Susan J.; Kizer, Jorge R.; Ix, Joachim H.; Mozaffarian, Dariush; Tracy, Russell P.; Psaty, Bruce M.; Siscovick, David S.; Mukamal, Kenneth J.; Djousse, Luc

    2014-01-01

    Plasma free fatty acids (FFA) are largely derived from adipose tissue. Elevated levels of FFA and fatty acid-binding protein 4 (FABP4), a key cytoplasmic chaperone of fatty acids, have been associated with adverse cardiovascular outcomes but limited data are available on the relation of these biomarkers with cardiovascular and total mortality. We studied 4,707 participants with a mean age of 75 years who had plasma FFA and FABP4 measured in 1992–1993 as part of the Cardiovascular Health Study, an observational cohort of community dwelling older adults. Over a median follow-up of 11.8 years, 3,555 participants died. Cox proportional hazard regression was used to determine the association between FFA, FABP4, and mortality. In fully adjusted models, FFA were associated with dose-dependent significantly higher total mortality (hazard ratio (HR) per standard deviation (SD): 1.14, 95% confidence interval (CI) 1.09–1.18), but FABP4 levels were not (HR 1.04, 95% CI 0.98–1.09). In a cause-specific mortality analysis, higher concentrations of FFA were associated with significantly higher risk of death due to cardiovascular disease, dementia, infection, and respiratory causes, but not cancer or trauma. We did not find evidence of an interaction between FFA and FABP4 (p=0.45), but FABP4 appeared to be associated with total mortality differentially among men and women (HR 1.17 (1.08–1.26) for men, HR 1.02 (0.96–1.07) for women, interaction p-value <0.001). In conclusion, in a cohort of community-dwelling older individuals, elevated plasma concentrations of FFA, but not FABP4, were associated with cardiovascular and non-cardiovascular mortality. PMID:25073566

  6. Impact of hyperlipidemia on plasma protein binding and hepatic drug transporter and metabolic enzyme regulation in a rat model of gestational diabetes.

    Science.gov (United States)

    Anger, Gregory J; Piquette-Miller, Micheline

    2010-07-01

    It is currently unknown whether gestational diabetes mellitus (GDM), a prevalent obstetrical complication, compounds the changes in drug disposition that occur naturally in pregnancy. Hyperlipidemia occurs in GDM. Using a rat model of GDM, we determined whether excess lipids compete with drugs for plasma protein binding. Because lipids activate nuclear receptors that regulate drug transporters and metabolic enzymes, we used proteome analysis to determine whether hyperlipidemia indirectly leads to the dysregulation of these proteins in the liver. GDM was induced on gestational day 6 (GD6) via streptozotocin injection. Controls received either vehicle alone or streptozotocin with subsequent insulin treatment. Liver and plasma were collected on GD20. Glyburide and saquinavir protein binding was determined by ultrafiltration, and an established solvent method was used for plasma delipidation. Proteomics analysis was performed by using isobaric tags for relative and absolute quantitation methodology with membrane-enriched hepatic protein samples. Relative to controls, GDM rat plasma contained more cholesterol and triglycerides. Plasma protein binding of glyburide and saquinavir was decreased in GDM. Delipidation normalized protein binding in GDM plasma. Proteins linked to lipid metabolism were strongly affected in the GDM proteomics data set, with prohyperlipidemic and antihyperlipidemic changes observed, and formed networks that implicated several nuclear receptors. Up-regulation of drug transporters and metabolic enzymes was observed (e.g., multidrug resistance 1/2, CYP2A1, CYP2B9, and CYP2D3). In this study, GDM-induced hyperlipidemia decreased protein binding and was associated with drug transporter and metabolic enzyme up-regulation in the liver. Both of these findings could change drug disposition in affected pregnancies, compounding changes associated with pregnancy itself.

  7. Plasma TNF binding capacity profiles during treatment with etanercept in rheumatoid arthritis

    DEFF Research Database (Denmark)

    Gudbrandsdottir, S; Bliddal, H; Petri, A;

    2004-01-01

    Etanercept (Enbrel) induces a rapid and sustained decline in disease activity in the majority of patients with refractory rheumatoid arthritis (RA). In these patients neutralization of TNFalpha and lymphotoxin (LT), previously termed TNFbeta is mediated by etanercept itself, as well as by naturally...... occurring soluble TNF receptors. However, the clinical response to treatment with etanercept may vary. Previously, pharmacokinetic studies have focused on the molar concentrations of etanercept, but very little is known about the kinetics of bioactive etanercept in patients treated with etanercept....... The purpose of this study was to evaluate kinetics, including inter- and intraindividual variations of the total TNF binding capacity, in RA patients who were on a standard treatment schedule with etanercept....

  8. The effect of organochlorines and heavy metals on sex steroid-binding proteins in vitro in the plasma of nesting green turtles, Chelonia mydas.

    Science.gov (United States)

    Ikonomopoulou, Maria Petrou; Olszowy, Henry; Hodge, Mary; Bradley, Adrian J

    2009-07-01

    In this study on green turtles, Chelonia mydas, from Peninsular Malaysia, the effect of selected environmental toxicants was examined in vitro. Emphasis was placed on purported hormone-mimicking chemicals such as dichlorodiphenyltrichloroethane (DDT), dichlorodiphenyldichloroethylene, dieldrin, lead, zinc and copper. Five concentrations were used: high (1 mg/L), medium (10(-1) mg/L), low (10(-2) mg/L), very low (10(-6) mg/L) and control (diluted carrier solvent but no toxicants). The results suggest that environmental pesticides and heavy metals may significantly alter the binding of steroids [i.e. testosterone (T) and oestradiol] to the plasma proteins in vitro. Competition studies showed that only Cu competed for binding sites with testosterone in the plasma collected from nesting C. mydas. Dieldrin and all heavy metals competed with oestradiol for binding sites. Furthermore, testosterone binding affinity was affected at various DDT concentrations and was hypothesised that DDT in vivo may act to inhibit steroid-protein interactions in nesting C. mydas. Although the precise molecular mechanism is yet to be described, DDT could have an effect upon the protein conformation thus affecting T binding (e.g. the T binding site on the steroid hormone binding protein molecule).

  9. Effects of exposure of epididymal boar spermatozoa to seminal plasma on the binding of zona pellucida proteins during in vitro capacitation

    NARCIS (Netherlands)

    Harkema, W.; Colenbrander, B.; Engel, B.; Woelders, H.

    2004-01-01

    The purpose of the investigation was to determine whether seminal plasma plays a role in the increase during in vitro capacitation of the number of boar spermatozoa with enhanced binding of zona pellucida proteins. Ejaculated spermatozoa and spermatozoa collected from the caudae epididymides of boar

  10. Deciphering the kinetic binding mechanism of dimeric ligands using a potent plasma-stable dimeric inhibitor of postsynaptic density protein-95 as an example

    DEFF Research Database (Denmark)

    Chi, Celestine N; Bach, Anders; Gottschalk, Marie

    2010-01-01

    addressed the kinetic mechanism of interaction of such bivalent ligands. We have investigated the binding interaction of a recently identified potent plasma-stable dimeric pentapeptide and PDZ1-2 of postsynaptic density protein-95 (PSD-95) using protein engineering in combination with fluorescence...

  11. Low abdominal NIRS values and elevated plasma intestinal fatty acid-binding protein in a premature piglet model of necrotizing enterocolitis

    Science.gov (United States)

    To identify early markers of necrotizing enterocolitis (NEC), we hypothesized that continuous abdominal near-infrared spectroscopy (A-NIRS) measurement of splanchnic tissue oxygen saturation and intermittent plasma intestinal fatty-acid binding protein (pI-FABP) measured every 6 hours can detect NEC...

  12. A High-Affinity Binding Site for the AVR9 Peptide Elicitor of Cladosporium fulvum Is Present on Plasma Membranes of Tomato and Other Solanaceous Plants.

    Science.gov (United States)

    Kooman-Gersmann, M.; Honee, G.; Bonnema, G.; De Wit, PJGM.

    1996-05-01

    The race-specific Cladosporium fulvum peptide elicitor AVR9, which specifically induces a hypersensitive response in tomato genotypes carrying the Cf-9 resistance gene, was labeled with iodine-125 at the N-terminal tyrosine residue and used in binding studies. 125I-AVR9 showed specific, saturable, and reversible binding to plasma membranes isolated from leaves of tomato cultivar Moneymaker without Cf resistance genes (MM-Cf0) or from a near-isogenic genotype with the Cf-9 resistance gene (MM-Cf9). The dissociation constant was found to be 0.07 nM, and the receptor concentration was 0.8 pmol/mg microsomal protein. Binding was highly influenced by pH and the ionic strength of the binding buffer and by temperature, indicating the involvement of both electrostatic and hydrophobic interactions. Binding kinetics and binding capacity were similar for membranes of the MM-Cf0 and MM-Cf9 genotypes. In all solanaceous plant species tested, an AVR9 binding site was present, whereas in the nonsolanaceous species that were analyzed, such a binding site could not be identified. The ability of membranes isolated from different solanaceous plant species to bind AVR9 seems to correlate with the presence of members of the Cf-9 gene family, but whether this correlation is functional remains to be determined.

  13. RNA-binding protein hnRNPLL regulates mRNA splicing and stability during B-cell to plasma-cell differentiation.

    Science.gov (United States)

    Chang, Xing; Li, Bin; Rao, Anjana

    2015-04-14

    Posttranscriptional regulation is a major mechanism to rewire transcriptomes during differentiation. Heterogeneous nuclear RNA-binding protein LL (hnRNPLL) is specifically induced in terminally differentiated lymphocytes, including effector T cells and plasma cells. To study the molecular functions of hnRNPLL at a genome-wide level, we identified hnRNPLL RNA targets and binding sites in plasma cells through integrated Photoactivatable-Ribonucleoside-Enhanced Cross-Linking and Immunoprecipitation (PAR-CLIP) and RNA sequencing. hnRNPLL preferentially recognizes CA dinucleotide-containing sequences in introns and 3' untranslated regions (UTRs), promotes exon inclusion or exclusion in a context-dependent manner, and stabilizes mRNA when associated with 3' UTRs. During differentiation of primary B cells to plasma cells, hnRNPLL mediates a genome-wide switch of RNA processing, resulting in loss of B-cell lymphoma 6 (Bcl6) expression and increased Ig production--both hallmarks of plasma-cell maturation. Our data identify previously unknown functions of hnRNPLL in B-cell to plasma-cell differentiation and demonstrate that the RNA-binding protein hnRNPLL has a critical role in tuning transcriptomes of terminally differentiating B lymphocytes.

  14. Concentrations of lipopolysaccharide-binding protein, bactericidal/permeability-increasing protein, soluble CD14 and plasma lipids in relation to endotoxaemia in patients with alcoholic liver disease

    DEFF Research Database (Denmark)

    Schäfer, C.; Parlesak, Alexandr; Schütt, C.;

    2002-01-01

    There is increasing evidence that gut leakage in persons with chronic alcohol misuse leads to endotoxaemia, which might contribute to the development of alcoholic hepatitis or cirrhosis. In addition, it was recently shown that the endotoxin-binding capacity of whole blood is reduced...... in these patients. To analyse this phenomenon, we measured the concentration of functionally important endotoxin-binding plasma components which modify the action of endotoxin. In patients with minimal (n = 10), intermediate (n = 9), and cirrhotic alcoholic liver disease (n = 11), and healthy controls (n = 11......), plasma endotoxin was determined in a limulus assay. The concentration of lipoproteins was assessed by measuring apolipoproteins, the other factors were directly measured in immunoassays. In the entire group of alcoholics, endotoxin and the concentration of binding factors that are involved in the action...

  15. Free and protein-bound insulin-like growth factor-I (IGF-I) and IGF-binding proteins in plasma of coho salmon, Oncorhynchus kisutch.

    Science.gov (United States)

    Shimizu, M; Swanson, P; Dickhoff, W W

    1999-09-01

    Total and free insulin-like growth factor-I (IGF-I) levels were quantified in plasma from growth hormone (GH)-treated and fasted coho salmon. Total IGF-I was measured by radioimmunoassay after acid-ethanol extraction and free IGF-I was separated from protein-bound IGF-I using ultrafiltration by centrifugation. Total and free IGF-I increased in plasma after GH treatment and decreased after fasting. The level of free IGF-I, however, was maintained at approximately 0.3% in both experiments. Unsaturated binding activity in plasma for IGF-I was assessed by incubation with (125)I-recombinant salmon IGF-I ((125)I-sIGF-I). Although there was no difference in binding activity between GH-treated and control fish, fasted fish showed higher binding activity than did fed fish, suggesting induction of unsaturated binding protein by fasting. IGF-binding protein (IGFBP) bands were observed in plasma of coho salmon by Western ligand blotting using (125)I-sIGF-I. A low-molecular-weight (22 kDa) band was clear in fasted fish but not detectable in fed fish. The IGFBP band, which has molecular weight similar to that of human IGFBP-3 (41 kDa), was more intense in GH-treated fish than in controls. The molecular distribution of IGF-I in plasma was examined by gel filtration under neutral conditions. Most IGF-I was eluted around 40 kDa. This result suggests that the major form of bound IGF-I in the circulation of coho salmon may be in a 40-kDa binary complex rather than in a 150-kDa ternary complex, as in mammals.

  16. A Polybasic Plasma Membrane Binding Motif in the I-II Linker Stabilizes Voltage-gated CaV1.2 Calcium Channel Function.

    Science.gov (United States)

    Kaur, Gurjot; Pinggera, Alexandra; Ortner, Nadine J; Lieb, Andreas; Sinnegger-Brauns, Martina J; Yarov-Yarovoy, Vladimir; Obermair, Gerald J; Flucher, Bernhard E; Striessnig, Jörg

    2015-08-21

    L-type voltage-gated Ca(2+) channels (LTCCs) regulate many physiological functions like muscle contraction, hormone secretion, gene expression, and neuronal excitability. Their activity is strictly controlled by various molecular mechanisms. The pore-forming α1-subunit comprises four repeated domains (I-IV), each connected via an intracellular linker. Here we identified a polybasic plasma membrane binding motif, consisting of four arginines, within the I-II linker of all LTCCs. The primary structure of this motif is similar to polybasic clusters known to interact with polyphosphoinositides identified in other ion channels. We used de novo molecular modeling to predict the conformation of this polybasic motif, immunofluorescence microscopy and live cell imaging to investigate the interaction with the plasma membrane, and electrophysiology to study its role for Cav1.2 channel function. According to our models, this polybasic motif of the I-II linker forms a straight α-helix, with the positive charges facing the lipid phosphates of the inner leaflet of the plasma membrane. Membrane binding of the I-II linker could be reversed after phospholipase C activation, causing polyphosphoinositide breakdown, and was accelerated by elevated intracellular Ca(2+) levels. This indicates the involvement of negatively charged phospholipids in the plasma membrane targeting of the linker. Neutralization of four arginine residues eliminated plasma membrane binding. Patch clamp recordings revealed facilitated opening of Cav1.2 channels containing these mutations, weaker inhibition by phospholipase C activation, and reduced expression of channels (as quantified by ON-gating charge) at the plasma membrane. Our data provide new evidence for a membrane binding motif within the I-II linker of LTCC α1-subunits essential for stabilizing normal Ca(2+) channel function. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  17. Effect of rosiglitazone on visfatin and retinol-binding protein-4 plasma concentrations in HIV-positive patients.

    Science.gov (United States)

    Haider, D G; Schindler, K; Mittermayer, F; Müller, M; Nowotny, P; Rieger, A; Luger, A; Ludvik, B; Wolzt, M

    2007-04-01

    Thiazolidinediones (TZD) may improve insulin resistance in patients with diabetes and HIV. The novel adipocytokines visfatin and retinol-binding protein-4 (RBP-4) have been proposed to influence the development of impaired glucose tolerance. The impact of TZD on these cytokines is yet unknown. In this randomized, double-blind, placebo-controlled parallel group study, 37 lean HIV-positive subjects aged 19-50 years were treated with 8 mg/day rosiglitazone (n=20) or placebo (n=17) for 6 months. Insulin sensitivity was estimated from the homeostasis model assessment (HOMA) index. Fasting visfatin, RBP-4, leptin, and adiponectin plasma concentrations were analyzed by immunoassays. Rosiglitazone had no effect on impaired insulin sensitivity, but increased median plasma visfatin from 6.2 ng/ml (95% CI: 5.9; 6.5) to 13.7 ng/ml (12.6; 19.1) (P<0.001) and adiponectin from 3.2 ng/ml (2.2; 4.0) to 4.0 ng/ml (3.3; 8.5; P<0.001). RBP-4 was lowered from 21.0 ng/ml (19.6; 23.1) to 16.3 ng/ml (15.2; 17.0; P<0.001), and leptin concentrations were unchanged. Adipocytokine concentrations were stable in subjects receiving placebo, where a deterioration in insulin sensitivity was detectable (P<0.05). Changes in visfatin and RBP-4 were correlated in subjects receiving rosiglitazone (r=-0.64, P<0.01) but not placebo (r=0.12, P=0.15). TZD treatment affects circulating adipocytokine concentrations in subjects with HIV. Reductions in RBP-4 and increases in visfatin may contribute to the pharmacodynamic action of TZD on glucose homeostasis. Quantification of adipocytokines might be useful to assess TZD treatment effectiveness in insulin-resistant subjects with HIV.

  18. An auxin-binding protein is localized to the plasma membrane of maize coleoptile cells: Identification by photoaffinity labeling and purification of a 23-kDa polypeptide

    Energy Technology Data Exchange (ETDEWEB)

    Feldwisch, J.; Zettl, R.; Hesse, F.; Schell, J.; Palme, K. (Max-Planck-Inst. fuer Zuechtungsforschung, Koeln (West Germany))

    1992-01-15

    Plasma membrane vesicles were isolated from maize (Zea mays L.) coleoptile tissue by aqueous two-phase partitioning and assayed for homogeneity by the use of membrane-specific enzymatic assays. Using 5-azido-(7-{sup 3}H)indole-3-acetic acid (({sup 3}H)N{sub 3}IAA), the authors identified several IAA-binding proteins with the molecular masses of 60 kDa (pm60), 58 kDa (pm58), and 23 kDa (pm23). Using Triton X-114, they were able to selectively extract pm23 from the plasma membrane. They show that auxins and functional analogues compete with ({sup 3}H)N{sub 3}IAA for binding to pm23. They found that PAB130, a polyclonal antibody raised against auxin-binding protein 1 (ABP-1), recognized ABP-1 as well as pm23. This suggests that pm23 shares common epitopes with ABP-1. In addition, they identified an auxin-binding protein with a molecular mass of 24 kDa (pm24), which was detected in microsomal but not in plasma membrane vesicle preparations. Like pm23 this protein was extracted from membrane vesicles with Triton X-114. They designed a purification scheme allowing simultaneous purification of pm23 and pm24. Homogeneous pm23 and pm24 were obtained from coleoptile extracts after 7,000-fold purification.

  19. Cell-penetrating compounds preferentially bind glycosaminoglycans over plasma membrane lipids in a charge density- and stereochemistry-dependent manner.

    Science.gov (United States)

    Prevette, Lisa E; Benish, Nicolas C; Schoenecker, Amber R; Braden, Kristin J

    2015-12-01

    Cell-penetrating compounds (CPCs) are often conjugated to drugs and genes to facilitate cellular uptake. We hypothesize that the electrostatic interaction between the positively charged amines of the cell-penetrating compounds and the negatively charged glycosaminoglycans (GAGs) extending from cell surfaces is the initiating step in the internalization process. The interactions of generation 5 PAMAM dendrimer, Tat peptide and 25 kDa linear PEI with four different GAGs have been studied using isothermal titration calorimetry to elucidate structure-function relationships that could lead to improved drug and gene delivery methods to a wide variety of cell types. Detailed thermodynamic analysis has determined that CPC-GAG binding constants range from 8.7×10(3) to 2.4×10(6)M(-1) and that affinity is dependent upon GAG charge density and stereochemistry and CPC molecular weight. The effect of GAG composition on affinity is likely due to hydrogen bonding between CPC amines and amides and GAG hydroxyl and amine groups. These results were compared to the association of CPCs with lipid vesicles of varying composition as model plasma membranes to finally clarify the relative importance of each cell surface component in initial cell recognition. CPC-lipid affinity increases with anionic lipid content, but GAG affinity is higher for all cell-penetrating compounds, confirming the role these heterogeneous polysaccharides play in cellular association and clustering.

  20. Humoral markers of active Epstein-Barr virus infection associate with anti-extractable nuclear antigen autoantibodies and plasma galectin-3 binding protein in systemic lupus erythematosus.

    Science.gov (United States)

    Rasmussen, N S; Nielsen, C T; Houen, G; Jacobsen, S

    2016-12-01

    We investigated if signs of active Epstein-Barr virus and cytomegalovirus infections associate with certain autoantibodies and a marker of type I interferon activity in patients with systemic lupus erythematosus. IgM and IgG plasma levels against Epstein-Barr virus early antigen diffuse and cytomegalovirus pp52 were applied as humoral markers of ongoing/recently active Epstein-Barr virus and cytomegalovirus infections, respectively. Plasma galectin-3 binding protein served as a surrogate marker of type I interferon activity. The measurements were conducted in 57 systemic lupus erythematosus patients and 29 healthy controls using ELISAs. Regression analyses and univariate comparisons were performed for associative evaluation between virus serology, plasma galectin-3 binding protein and autoantibodies, along with other clinical and demographic parameters. Plasma galectin-3 binding protein concentrations were significantly higher in systemic lupus erythematosus patients (P = 0.009) and associated positively with Epstein-Barr virus early antigen diffuse-directed antibodies and the presence of autoantibodies against extractable nuclear antigens in adjusted linear regressions (B = 2.02 and 2.02, P = 0.02 and P = 0.002, respectively). Furthermore, systemic lupus erythematosus patients with anti-extractable nuclear antigens had significantly higher antibody levels against Epstein-Barr virus early antigen diffuse (P = 0.02). Our study supports a link between active Epstein-Barr virus infections, positivity for anti-extractable nuclear antigens and increased plasma galectin-3 binding protein concentrations/type I interferon activity in systemic lupus erythematosus patients. © The Author(s) 2016.

  1. Treponema pallidum subsp. pallidum TP0136 protein is heterogeneous among isolates and binds cellular and plasma fibronectin via its NH2-terminal end.

    Science.gov (United States)

    Ke, Wujian; Molini, Barbara J; Lukehart, Sheila A; Giacani, Lorenzo

    2015-03-01

    Adherence-mediated colonization plays an important role in pathogenesis of microbial infections, particularly those caused by extracellular pathogens responsible for systemic diseases, such as Treponema pallidum subsp. pallidum (T. pallidum), the agent of syphilis. Among T. pallidum adhesins, TP0136 is known to bind fibronectin (Fn), an important constituent of the host extracellular matrix. To deepen our understanding of the TP0136-Fn interaction dynamics, we used two naturally-occurring sequence variants of the TP0136 protein to investigate which region of the protein is responsible for Fn binding, and whether TP0136 would adhere to human cellular Fn in addition to plasma Fn and super Fn as previously reported. Fn binding assays were performed with recombinant proteins representing the two full-length TP0136 variants and their discrete regions. As a complementary approach, we tested inhibition of T. pallidum binding to Fn by recombinant full-length TP0136 proteins and fragments, as well as by anti-TP0136 immune sera. Our results show that TP0136 adheres more efficiently to cellular Fn than to plasma Fn, that the TP0136 NH2-terminal conserved region of the protein is primarily responsible for binding to plasma Fn but that binding sites for cellular Fn are also present in the protein's central and COOH-terminal regions. Additionally, message quantification studies show that tp0136 is highly transcribed during experimental infection, and that its message level increases in parallel to the host immune pressure on the pathogen, which suggests a possible role for this protein in T. pallidum persistence. In a time where syphilis incidence is high, our data will help in the quest to identify suitable targets for development of a much needed vaccine against this important disease.

  2. Plasma Ang2 and ADAM17 levels are elevated during clinical malaria; Ang2 level correlates with severity and expression of EPCR-binding PfEMP1

    DEFF Research Database (Denmark)

    Petersen, Jens E V; Mkumbaye, Sixbert I; Vaaben, Anna V;

    2016-01-01

    Membrane Protein 1 (PfEMP1) containing Cysteine Rich Inter Domain Region α1 (CIDRα1) domains predicted to bind Endothelial Protein C receptor (EPCR). ADAM17 levels were not associated with expression of var genes encoding different PfEMP1 types when controlling for age. These data are the first to report...... ADAM17 plasma levels in malaria-exposed individuals, and support the notion that parasite sequestration mediated by EPCR-binding PfEMP1 is associated with endothelial activation and pathology in severe paediatric malaria....

  3. Plasma Levels of Fatty Acid-Binding Protein 4, Retinol-Binding Protein 4, High-Molecular-Weight Adiponectin, and Cardiovascular Mortality Among Men With Type 2 Diabetes: A 22-Year Prospective Study.

    Science.gov (United States)

    Liu, Gang; Ding, Ming; Chiuve, Stephanie E; Rimm, Eric B; Franks, Paul W; Meigs, James B; Hu, Frank B; Sun, Qi

    2016-11-01

    To examine select adipokines, including fatty acid-binding protein 4, retinol-binding protein 4, and high-molecular-weight (HMW) adiponectin in relation to cardiovascular disease (CVD) mortality among patients with type 2 diabetes mellitus. Plasma levels of fatty acid-binding protein 4, retinol-binding protein 4, and HMW adiponectin were measured in 950 men with type 2 diabetes mellitus in the Health Professionals Follow-up Study. After an average of 22 years of follow-up (1993-2015), 580 deaths occurred, of whom 220 died of CVD. After multivariate adjustment for covariates, higher levels of fatty acid-binding protein 4 were significantly associated with a higher CVD mortality: comparing extreme tertiles, the hazard ratio and 95% confidence interval of CVD mortality was 1.78 (1.22-2.59; P trend=0.001). A positive association was also observed for HMW adiponectin: the hazard ratio (95% confidence interval) was 2.07 (1.42-3.06; P trend=0.0002), comparing extreme tertiles, whereas higher retinol-binding protein 4 levels were nonsignificantly associated with a decreased CVD mortality with an hazard ratio (95% confidence interval) of 0.73 (0.50-1.07; P trend=0.09). A Mendelian randomization analysis suggested that the causal relationships of HMW adiponectin and retinol-binding protein 4 would be directionally opposite to those observed based on the biomarkers, although none of the Mendelian randomization associations achieved statistical significance. These data suggest that higher levels of fatty acid-binding protein 4 and HMW adiponectin are associated with elevated CVD mortality among men with type 2 diabetes mellitus. Biological mechanisms underlying these observations deserve elucidation, but the associations of HMW adiponectin may partially reflect altered adipose tissue functionality among patients with type 2 diabetes mellitus. © 2016 American Heart Association, Inc.

  4. Molecular Properties of Guar Gum and Pectin Modify Cecal Bile Acids, Microbiota, and Plasma Lipopolysaccharide-Binding Protein in Rats.

    Science.gov (United States)

    Ghaffarzadegan, Tannaz; Marungruang, Nittaya; Fåk, Frida; Nyman, Margareta

    2016-01-01

    Bile acids (BAs) act as signaling molecules in various physiological processes, and are related to colonic microbiota composition as well as to different types of dietary fat and fiber. This study investigated whether guar gum and pectin-two fibers with distinct functional characteristics-affect BA profiles, microbiota composition, and gut metabolites in rats. Low- (LM) or high-methoxylated (HM) pectin, and low-, medium-, or high-molecular-weight (MW) guar gum were administered to rats that were fed either low- or high-fat diets. Cecal BAs, short-chain fatty acids (SCFA) and microbiota composition, and plasma lipopolysaccharide-binding protein (LBP) levels were analyzed, by using novel methodologies based on gas chromatography (BAs and SCFAs) and 16S rRNA gene sequencing on the Illumina MiSeq platform. Strong correlations were observed between cecal BA and SCFA levels, microbiota composition, and portal plasma LBP levels in rats on a high-fat diet. Notably, guar gum consumption with medium-MW increased the cecal amounts of cholic-, chenodeoxycholic-, and ursodeoxycholic acids as well as α-, β-, and ω-muricholic acids to a greater extent than other types of guar gum or the fiber-free control diet. In contrast, the amounts of cecal deoxycholic- and hyodeoxycholic acid were reduced with all types of guar gum independent of chain length. Differences in BA composition between pectin groups were less obvious, but cecal levels of α- and ω-muricholic acids were higher in rats fed LM as compared to HM pectin or the control diet. The inflammatory marker LBP was downregulated in rats fed medium-MW guar gum and HM pectin; these two fibers decreased the cecal abundance of Oscillospira and an unclassified genus in Ruminococcaceae, and increased that of an unclassified family in RF32. These results indicate that the molecular properties of guar gum and pectin are important for their ability to modulate cecal BA formation, gut microbiota composition, and high-fat diet induced

  5. Study on Plasma Protein Binding Rate of Hesperetin in Rats%橙皮素的大鼠血浆蛋白结合率研究

    Institute of Scientific and Technical Information of China (English)

    吴中杰; 张艳; 胡佳骅

    2015-01-01

    目的:研究橙皮素的大鼠血浆蛋白结合率。方法使用平衡透析法模拟橙皮素在体内与血浆蛋白结合的过程,采用高效液相色谱法(HPLC)测定低、中、高3个浓度(0.5,2.5和10μg/ml)的橙皮素在透析袋内血浆中和透析袋外缓冲液中的浓度,计算血浆蛋白结合率。结果低、中、高3个浓度的橙皮素在体外与大鼠血浆蛋白的平均结合率分别为83.5%,84.3%和83.9%,3个浓度之间的血浆蛋白结合率比较,差异无统计学意义( P ﹥0.05)。结论橙皮素具有较高的血浆蛋白结合率,为含橙皮苷药物的临床合理应用提供理论指导。%Objective To study the plasma protein binding rate of hesperetin in rats. Methods The equilibrium dialysis method was used to imitate the in vivo plasma protein binding process of hesperetin. HPLC was adopted to determine the percentages of hesperetin in the plasma of dialysis bag and in the buffer out of the dialysis bag at low,middle and high concentrations(0. 5,2. 5 and10 μg/ ml). Results The aver-age plasma protein binding rates were 83. 5% ,84. 3% and 83. 9% in vitro at the low,middle and high con-centrations of hesperetin. The difference in plasma protein binding rate was not significant among the three concentrations(P ﹥ 0. 05). Conclusion Hesperetin has very high plasma protein binding rate,which pro-vides the theoretic guidance for the clinical rational application of hesperidin drugs.

  6. Exposure to positively- and negatively-charged plasma cluster ions impairs IgE-binding capacity of indoor cat and fungal allergens

    OpenAIRE

    NISHIKAWA, Kazuo; Fujimura, Takashi; Ota, Yasuhiro; Abe, Takuya; ElRamlawy, Kareem Gamal; Nakano, Miyako; Takado, Tomoaki; Uenishi, Akira; Kawazoe, Hidechika; Sekoguchi, Yoshinori; Tanaka, Akihiko; Ono, Kazuhisa; Kawamoto, Seiji

    2016-01-01

    Background Environmental control to reduce the amount of allergens in a living place is thought to be important to avoid sensitization to airborne allergens. However, efficacy of environmental control on inactivation of airborne allergens is not fully investigated. We have previously reported that positively- and negatively-charged plasma cluster ions (PC-ions) reduce the IgE-binding capacity of crude allergens from Japanese cedar pollen as important seasonal airborne allergens. Cat (Felis do...

  7. Stability of the tumor suppressor merlin depends on its ability to bind paxillin LD3 and associate with β1 integrin and actin at the plasma membrane

    Directory of Open Access Journals (Sweden)

    Maria Elisa Manetti

    2012-08-01

    The NF2 gene encodes a tumor suppressor protein known as merlin or schwannomin whose loss of function causes Neurofibromatosis Type 2 (NF2. NF2 is characterized by the development of benign tumors, predominantly schwannomas, in the peripheral nervous system. Merlin links plasma membrane receptors with the actin cytoskeleton and its targeting to the plasma membrane depends on direct binding to the paxillin scaffold protein. Exon 2 of NF2, an exon mutated in NF2 patients and deleted in a mouse model of NF2, encodes the merlin paxillin binding domain (PBD1. Here, we sought to determine the role of PBD1 in regulation of merlin stability and association with plasma membrane receptors and the actin cytoskeleton in Schwann cells. Using a fluorescence-based pulse-chase technique, we measured the half-life of Halo-tagged merlin variants carrying PBD1, exon 2, and exons 2 and 3 deletions in transiently transfected Schwann cells. We found that PBD1 alone was necessary and sufficient to increase merlin's half-life from approximately three to eleven hours. Merlin lacking PBD1 did not form a complex with surface β1 integrins or associate with the actin cytoskeleton. In addition, direct binding studies using purified merlin and paxillin domains revealed that merlin directly binds paxillin LD3 (leucine-aspartate 3 domain as well as the LD4 and LD5 domains. Together these results demonstrate that a direct interaction between merlin PBD1 and the paxillin LD3–5 domains targets merlin to the plasma membrane where it is stabilized by its association with surface β1 integrins and cortical actin.

  8. Effects of competitive red blood cell binding and reduced hematocrit on the blood and plasma levels of (/sup 14/C)Indapamide in the rat

    Energy Technology Data Exchange (ETDEWEB)

    Lettieri, J.T.; Portelli, S.T.

    1983-02-01

    The effects of chlorthalidone and acetazolamide on the red blood cell binding of indapamide were investigated. Both drugs caused a substantial decrease in the amount of indapamide bound to the erythrocytes in vitro. This effect was demonstrated by a change in the indapamide blood/plasma ratio from approximately 6 in control samples, to a value of 1 when either of the displacing agents was added. Coadministration of acetazolamide with /sup 14/C-labeled indapamide to rats, resulted in a 5-fold drop in the blood levels of total radioactivity, relative to rats dosed with (/sup 14/C)indapamide alone. Concomitantly, there was a 2-fold increase in the plasma levels of total radioactivity after acetazolamide coadministration. In rats whose hematocrits had been reduced by extensive bleeding, there were only minor alterations in the blood/plasma partitioning of (/sup 14/C)indapamide. Thus, chlorthalidone and acetazolamide were able to displace indapamide from erythrocytes in vitro and in vivo, possibly by competition at a carbonic anhydrase binding site. The pharmacokinetics of drugs which are extensively bound to erythrocytes may be significantly altered by the presence of other agents capable of competitive binding.

  9. Deglycosylation of serum vitamin D3-binding protein by alpha-N-acetylgalactosaminidase detected in the plasma of patients with systemic lupus erythematosus.

    Science.gov (United States)

    Yamamoto, N; Naraparaju, V R; Moore, M; Brent, L H

    1997-03-01

    A serum glycoprotein, Gc protein (vitamin D3-binding protein), can be converted by beta-galactosidase of B cells and sialidase of T cells to a potent macrophage-activating factor (MAF), a protein with N-acetylgalactosamine as the remaining sugar moiety. Thus, Gc protein is the precursor for MAF. Treatment of Gc protein with immobilized beta-galactosidase and sialidase generates a remarkably high titered macrophage-activating factor (GcMAF). When peripheral blood monocytes/ macrophages (designated macrophages) of 33 systemic lupus erythematosus patients were incubated with GcMAF (100 pg/ml), the macrophages of all patients were activated as determined by superoxide generation. However, the precursor activity of patient plasma Gc protein was lost or reduced in these patients. Loss of the precursor activity was the result of deglycosylation of plasma Gc protein by alpha-N-acetylgalactosaminidase activity found in the patient plasma. Levels of plasma alpha-N-acetylgalactosaminidase activity in individual patients had an inverse correlation with the MAF precursor activity of their plasma Gc protein. Deglycosylated Gc protein cannot be converted to macro-phage-activating factor. The resulting defect in macro-phage activation may lead to an inability to clear pathogenic immune complexes. Thus, elevated plasma alpha-N-acetylgalactosaminidase activity resulting in the loss of MAF precursor activity and reduced macro-phage activity may play a role in the pathogenesis of systemic lupus erythematosus.

  10. Increased plasma concentrations of vitamin D metabolites and vitamin D binding protein in women using hormonal contraceptives: a cross-sectional study

    DEFF Research Database (Denmark)

    Liendgaard, Ulla Kristine Møller; við Streym, Susanna; Jensen, Lars Thorbjørn

    2013-01-01

    UNLABELLED: Use of hormonal contraceptives (HC) may influence total plasma concentrations of vitamin D metabolites. A likely cause is an increased synthesis of vitamin D binding protein (VDBP). Discrepant results are reported on whether the use of HC affects free concentrations of vitamin D...... winter season. Compared with non-users (n = 23), users of HC (n = 52) had significantly higher plasma concentrations of 25-hydroxyvitamin D (25OHD) (median 84 interquartile range: [67-111] vs. 70 [47-83] nmol/L, p = 0.01), 1,25-dihydroxyvitamin D (1,25(OH)2D) (198 [163-241] vs. 158 [123-183] pmol/L, p...... = 0.01) and VDBP (358 [260-432] vs. 271 [179-302] µg/mL, p OH)2D) were not significantly different between groups (p > 0.10). There were no significant differences in indices of calcium homeostasis (plasma concentrations of calcium...

  11. Surface mapping of binding of oviductin to the plasma membrane of golden hamster spermatozoa during in vitro capacitation and acrosome reaction.

    Science.gov (United States)

    Kan, Frederick W K; Esperanzate, Patrick W B

    2006-06-01

    Oviductins are high-molecular-weight glycoproteins synthesized and secreted by nonciliated oviductal epithelial cells and have been shown to play a role in fertilization and early embryo development. The present study was carried out to examine the in vitro binding capacity of hamster oviductin to homologous sperm and to determine the sites of its localization in untreated, capacitated, and acrosome-reacted spermatozoa. Freshly prepared epididymal and capacitated sperm as well as acrosome-reacted sperm were incubated with oviductal fluid prepared from isolated hamster oviducts, fixed and then probed with a monoclonal antibody against hamster oviductin. Results obtained with pre-embedding immunolabeling experiments revealed binding of oviductin to the acrosomal cap and the apical aspect of the postacrosomal region. Immunolabeling of both regions appeared to be more intense in capacitated spermatozoa. Acrosome-reacted sperm showed an immunoreaction of moderate intensity over the postacrosomal region. The plasma membrane overlying the equatorial segment also exhibited a weak labeling. Quantitative analysis obtained with the surface replica technique indicated that oviductin had a higher binding affinity for the acrosomal cap than the postacrosomal region and that the binding of oviductin to the latter plasma membrane domain was enhanced during capacitation. Binding of oviductin to the postacrosomal region, however, was attenuated after acrosome reaction. Immunolabeling for oviductin was found to be the weakest over the equatorial segment regardless of the experimental conditions. The binding of hamster oviductin to specific membrane domains of the homologous sperm and the changes in its distribution during capacitation and acrosome reaction may be important for the function of hamster oviductin preceding and during fertilization.

  12. Relation between retinol, retinol-binding protein 4, transthyretin and carotid intima media thickness.

    Science.gov (United States)

    Bobbert, Thomas; Raila, Jens; Schwarz, Franziska; Mai, Knut; Henze, Andrea; Pfeiffer, Andreas F H; Schweigert, Florian J; Spranger, Joachim

    2010-12-01

    Retinol is transported in a complex with retinol-binding protein 4 (RBP4) and transthyretin (TTR) in the circulation. While retinol is associated with various cardiovascular risk factors, the relation between retinol, RBP4, TTR and carotid intima media thickness (IMT) has not been analysed yet. Retinol, RBP4 and TTR were measured in 96 individuals and their relation to mean and maximal IMT was determined. Mean IMT correlated with RBP4 (r=0.335, p<0.001), retinol (r=-0.241, p=0.043), RBP/TTR ratio (r=0.254, p=0.025) and retinol/RBP4 ratio (r=-0.549, p<0.001). Adjustment for age, sex, BMI, blood pressure, HDL/total cholesterol ratio, triglyceride, diabetes and smoking revealed that the retinol/RBP4 ratio was strongly and independently associated with mean IMT. Similar results were found for maximal IMT, which included the measurement of plaques. The data support that the transport complex of vitamin A is associated with the IMT, an established parameter of atherosclerosis. Changes in RBP4 saturation with retinol may link renal dysfunction and insulin resistance to atherosclerosis. Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.

  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.

    Science.gov (United States)

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

    2013-06-27

    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. Regulation of formyl peptide receptor binding to rabbit neutrophil plasma membranes. Use of monovalent cations, guanine nucleotides, and bacterial toxins to discriminate among different states of the receptor

    Energy Technology Data Exchange (ETDEWEB)

    Feltner, D.E.; Marasco, W.A.

    1989-06-01

    The regulation by monovalent cations, guanine nucleotides, and bacterial toxins of (3H)FMLP binding to rabbit neutrophil plasma membranes was studied by using dissociation techniques to identify regulatory effects on separate receptor states. Under conditions of low receptor occupancy (1 nM (3H)FMLP) and in both Na+ and K+ buffers, dissociation is heterogenous, displaying two distinct, statistically significant off rates. (3H)FMLP binding was enhanced by substituting other monovalent cations for Na+. In particular, enhanced binding in the presence of K+ relative to Na+ was caused by additional binding to both rapidly and slowly dissociating receptors. Three receptor dissociation rates, two of which appear to correspond to the two affinity states detected in equilibrium binding studies, were defined by specific GTP and pertussis toxin (PT) treatments. Neither GTP, nor PT or cholera toxins (CT) had an effect on the rate of dissociation of (3H)FMLP from the rapidly dissociating form of the receptor. Both 100 microM GTP and PT treatments increased the percentage of rapidly dissociating receptors, correspondingly decreasing the percentage of slowly dissociating receptors. The observed changes in the rapidly and slowly dissociating receptors after GTP, PT, and CT treatments were caused by an absolute decrease in the amount of binding to the slowly dissociating receptors. However, complete inhibition of slowly dissociating receptor binding by GTP, PT, or both was never observed. Both GTP and PT treatments, but not CT treatment, increased by two-fold the rate of dissociation of 1 nM (3H)FMLP from the slowly dissociating form of the receptor, resulting in a third dissociation rate. Thus, slowly dissociating receptors comprise two different receptor states, a G protein-associated guanine nucleotide and PT-sensitive state and a guanine nucleotide-insensitive state.

  15. Zinc-binding proteins from boar seminal plasma -- isolation, biochemical characteristics and influence on spermatozoa stored at 4°C.

    Science.gov (United States)

    Mogielnicka-Brzozowska, Marzena; Wysocki, Paweł; Strzeżek, Jerzy; Kordan, Władysław

    2011-01-01

    Affinity chromatography on Chelating Sepharose Fast Flow Gel-Zn(2+) was used for fractionation of boar seminal plasma proteins. Approximately 30% of total boar seminal plasma proteins showed affinity for zinc ions (ZnBP fraction). Native electrophoresis (PAGE) of ZnBP revealed six protein fractions which separated into 27 bands under denaturing conditions (SDS/PAGE). Two-dimensional electrophoresis (2D PAGE) showed 148 polypeptides with isoelectric points mostly in the basic and neutral pH range. The zinc-binding proteins comprise mainly 10-20 kDa polypeptides which are probably members of the spermadhesin family. ZnBP present in the incubation mixture of spermatozoa stored for 1 or 24 h at 4 °C allowed preservation of a higher percentage of cells exhibiting linear motility in comparison to a control sample stored in PBS. Presented results indicate that proteins binding Zn(2+) ions have a shielding effect on the sperm plasma membrane and acrosome of spermatozoa, protecting these structures against consequences of cold shock.

  16. Binding of Alphaherpesvirus Glycoprotein H to Surface α4β1-Integrins Activates Calcium-Signaling Pathways and Induces Phosphatidylserine Exposure on the Plasma Membrane

    Science.gov (United States)

    Gramatica, Andrea; Herrmann, Andreas; Osterrieder, Nikolaus

    2015-01-01

    ABSTRACT Intracellular signaling connected to integrin activation is known to induce cytoplasmic Ca2+ release, which in turn mediates a number of downstream signals. The cellular entry pathways of two closely related alphaherpesviruses, equine herpesviruses 1 and 4 (EHV-1 and EHV-4), are differentially regulated with respect to the requirement of interaction of glycoprotein H (gH) with α4β1-integrins. We show here that binding of EHV-1, but not EHV-4, to target cells resulted in a rapid and significant increase in cytosolic Ca2+ levels. EHV-1 expressing EHV-4 gH (gH4) in lieu of authentic gH1 failed to induce Ca2+ release, while EHV-4 with gH1 triggered significant Ca2+ release. Blocking the interaction between gH1 and α4β1-integrins, inhibiting phospholipase C (PLC) activation, or blocking binding of inositol 1,4,5-triphosphate (IP3) to its receptor on the endoplasmic reticulum (ER) abrogated Ca2+ release. Interestingly, phosphatidylserine (PS) was exposed on the plasma membrane in response to cytosolic calcium increase after EHV-1 binding through a scramblase-dependent mechanism. Inhibition of both Ca2+ release from the ER and scramblase activation blocked PS scrambling and redirected virus entry to the endocytic pathway, indicating that PS may play a role in facilitating virus entry directly at the plasma membrane. PMID:26489864

  17. Genome wide association identifies common variants at the SERPINA6/SERPINA1 locus influencing plasma cortisol and corticosteroid binding globulin

    NARCIS (Netherlands)

    Bolton, Jennifer L.; Hayward, Caroline; Direk, Nese; Lewis, John G.; Hammond, Geoffrey L.; Hill, Lesley A.; Anderson, Anna; Huffman, Jennifer; Wilson, James F.; Campbell, Harry; Rudan, Igor; Wright, Alan; Hastie, Nicholas; Wild, Sarah H.; Velders, Fleur P.; Hofman, Albert; Uitterlinden, Andre G.; Lahti, Jari; Raikkonen, Katri; Kajantie, Eero; Widen, Elisabeth; Palotie, Aarno; Eriksson, Johan G.; Kaakinen, Marika; Jarvelin, Marjo-Riitta; Timpson, Nicholas J.; Smith, George Davey; Ring, Susan M.; Evans, David M.; St Pourcain, Beate; Tanaka, Toshiko; Milaneschi, Yuri; Bandinelli, Stefania; Ferrucci, Luigi; van der Harst, Pim; Rosmalen, Judith G. M.; Bakker, Stephen J. L.; Verweij, Niek; Dullaart, Robin P. F.; Mahajan, Anubha; Lindgren, Cecilia M.; Morris, Andrew; Lind, Lars; Ingelsson, Erik; Anderson, Laura N.; Pennell, Craig E.; Lye, Stephen J.; Matthews, Stephen G.; Eriksson, Joel; Mellstrom, Dan; Ohlsson, Claes; Price, Jackie F.; Strachan, Mark W. J.; Reynolds, Rebecca M.; Tiemeier, Henning; Walker, Brian R.

    Variation in plasma levels of cortisol, an essential hormone in the stress response, is associated in population-based studies with cardio-metabolic, inflammatory and neuro-cognitive traits and diseases. Heritability of plasma cortisol is estimated at 30-60% but no common genetic contribution has

  18. Streamlined Approach for Environmental Restoration (SAFER) Plan for Corrective Action Unit 408: Bomblet Target Area Tonopah Test Range (TTR), Nevada, Revision 1

    Energy Technology Data Exchange (ETDEWEB)

    Mark Krauss

    2010-03-01

    This Streamlined Approach for Environmental Restoration Plan addresses the actions needed to achieve closure of Corrective Action Unit (CAU) 408, Bomblet Target Area (TTR). Corrective Action Unit 408 is located at the Tonopah Test Range and is currently listed in Appendix III of the Federal Facility Agreement and Consent Order. Corrective Action Unit 408 comprises Corrective Action Site TA-55-002-TAB2, Bomblet Target Areas. Clean closure of CAU 408 will be accomplished by removal of munitions and explosives of concern within seven target areas and potential disposal pits. The target areas were used to perform submunitions related tests for the U.S. Department of Energy (DOE). The scope of CAU 408 is limited to submunitions released from DOE activities. However, it is recognized that the presence of other types of unexploded ordnance and munitions may be present within the target areas due to the activities of other government organizations. The CAU 408 closure activities consist of: • Clearing bomblet target areas within the study area. • Identifying and remediating disposal pits. • Collecting verification samples. • Performing radiological screening of soil. • Removing soil containing contaminants at concentrations above the action levels. Based on existing information, contaminants of potential concern at CAU 408 include unexploded submunitions, explosives, Resource Conservation Recovery Act metals, and depleted uranium. Contaminants are not expected to be present in the soil at concentrations above the action levels; however, this will be determined by radiological surveys and verification sample results.

  19. Isolation and Identification of Concanavalin A Binding Glycoproteins from Human Seminal Plasma: A Step Towards Identification of Male Infertility Marker Proteins

    Directory of Open Access Journals (Sweden)

    Anil Kumar Tomar

    2011-01-01

    Full Text Available Human seminal plasma contains a large array of proteins of clinical importance which are essentially needed to maintain the reproductive physiology of spermatozoa and for successful fertilization. Thus, isolation and identification of seminal plasma proteins is of paramount significance for their biophysical characterization and functional analysis in reproductive physiological processes. In this study, we have isolated Concanavalin-A binding glycoproteins from human seminal plasma and subsequently identified them by MALDI-TOF/MS analysis. The major proteins, as identified in this study, are Aminopeptidase N, lactoferrin, prostatic acid phosphatase, zinc-alpha-2-glycoprotein, prostate specific antigen, progestagen-associated endometrial protein, Izumo sperm-egg fusion protein and prolactin inducible protein. This paper also reports preliminary studies to identify altered expression of these proteins in oligospermia and azoospermia in comparison to normospermia. In oligospermia, five proteins were found to be downregulated while in azoospermia, four proteins were downregulated and two proteins were upregulated. Thus, this study is of immense biomedical interest towards identification of potential male infertility marker proteins in seminal plasma.

  20. Isolation and Identification of Concanavalin A Binding Glycoproteins from Human Seminal Plasma: A Step Towards Identification of Male Infertility Marker Proteins

    Science.gov (United States)

    Tomar, Anil Kumar; Sooch, Balwinder Singh; Raj, Isha; Singh, Sarman; Singh, Tej P.; Yadav, Savita

    2011-01-01

    Human seminal plasma contains a large array of proteins of clinical importance which are essentially needed to maintain the reproductive physiology of spermatozoa and for successful fertilization. Thus, isolation and identification of seminal plasma proteins is of paramount significance for their biophysical characterization and functional analysis in reproductive physiological processes. In this study, we have isolated Concanavalin-A binding glycoproteins from human seminal plasma and subsequently identified them by MALDI-TOF/MS analysis. The major proteins, as identified in this study, are Aminopeptidase N, lactoferrin, prostatic acid phosphatase, zinc-alpha-2-glycoprotein, prostate specific antigen, progestagen-associated endometrial protein, Izumo sperm-egg fusion protein and prolactin inducible protein. This paper also reports preliminary studies to identify altered expression of these proteins in oligospermia and azoospermia in comparison to normospermia. In oligospermia, five proteins were found to be downregulated while in azoospermia, four proteins were downregulated and two proteins were upregulated. Thus, this study is of immense biomedical interest towards identification of potential male infertility marker proteins in seminal plasma. PMID:22182811

  1. 3T3 fibroblasts transfected with a cDNA for mitochondrial aspartate aminotransferase express plasma membrane fatty acid-binding protein and saturable fatty acid uptake.

    OpenAIRE

    1995-01-01

    To explore the relationship between mitochondrial aspartate aminotransferase (mAspAT; EC 2.6.1.1) and plasma membrane fatty acid-binding protein (FABPpm) and their role in cellular fatty acid uptake, 3T3 fibroblasts were cotransfected with plasmid pMAAT2, containing a full-length mAspAT cDNA downstream of a Zn(2+)-inducible metallothionein promoter, and pFR400, which conveys methotrexate resistance. Transfectants were selected in methotrexate, cloned, and exposed to increasing methotrexate co...

  2. Study on plasma protein binding rate of naringin in rats%柚皮苷的大鼠血浆蛋白结合率研究

    Institute of Scientific and Technical Information of China (English)

    李季泓

    2015-01-01

    目的:研究柚皮苷的大鼠血浆蛋白结合率。方法使用平衡透析法模拟柚皮苷在体内与血浆蛋白结合的过程,并建立测定柚皮苷的高效液相色谱-质谱联用( HPLC-MS/MS)方法,测定低、中、高3个浓度(100、600、3600 ng/mL)的柚皮苷在透析袋内血浆中和透析袋外缓冲液中的浓度,计算血浆蛋白结合率。结果低、中、高3个浓度的柚皮苷在体外与大鼠的平均血浆蛋白结合率分别为91.5%、90.5%和96.3%,3个浓度之间的血浆蛋白结合率比较差异无统计学意义(P>0.05)。结论柚皮苷与大鼠血浆蛋白高度结合。%Objective To study the plasma protein binding rate of naringin in rats. Methods Under three dif-ferent concentrations (100,600 and 3 600 ng/mL),this paper used equilibrium dialysis method to imitate the binding process between naringin and the plasma protein of rats,and determined the concentration of naringin in and out of the dialysis membrane by high performance liquid chromatography-tandem mass spectrometry ( LC-MS/MS) method. The binding rate was calculated. Results Under the experimental concentrations,the plasma protein binding rates of naring-in in rats were 91. 5%,90. 5% and 96. 3% at low,middle and high concentrations,respectively. There was no signifi-cant difference among the three concentrations. Conclusion Naringin is highly bound to plasma proteins in rats.

  3. Inositol phospholipids regulate the guanine-nucleotide-exchange factor Tiam1 by facilitating its binding to the plasma membrane and regulating GDP/GTP exchange on Rac1.

    Science.gov (United States)

    Fleming, Ian N; Batty, Ian H; Prescott, Alan R; Gray, Alex; Kular, Gursant S; Stewart, Hazel; Downes, C Peter

    2004-09-15

    Binding of the Rac1-specific guanine-nucleotide-exchange factor, Tiam1, to the plasma membrane requires the N-terminal pleckstrin homology domain. In the present study, we show that membrane-association is mediated by binding of PtdIns(4,5)P(2) to the pleckstrin homology domain. Moreover, in 1321N1 astrocytoma cells, translocation of Tiam1 to the cytosol, following receptor-mediated stimulation of PtdIns(4,5)P(2) breakdown, correlates with decreased Rac1-GTP levels, indicating that membrane-association is required for GDP/GTP exchange on Rac1. In addition, we show that platelet-derived growth factor activates Rac1 in vivo by increasing PtdIns(3,4,5)P(3) concentrations, rather than the closely related lipid, PtdIns(3,4)P(2). Finally, the data demonstrate that PtdIns(4,5)P(2) and PtdIns(3,4,5)P(3) bind to the same pleckstrin homology domain in Tiam1 and that soluble inositol phosphates appear to compete with lipids for this binding. Together, these novel observations provide strong evidence that distinct phosphoinositides regulate different functions of this enzyme, indicating that local concentrations of signalling lipids and the levels of cytosolic inositol phosphates will play crucial roles in determining its activity in vivo.

  4. Interaction of xenobiotics with estrogen receptors α and β and a putative plasma sex hormone-binding globulin from channel catfish (Ictalurus punctatus)

    Science.gov (United States)

    Gale, William L.; Patino, Reynaldo; Maule, Alec G.

    2004-01-01

    Estrogens are important regulators of physiological functions. Although environmental contaminants (xenoestrogens) which interfere with estrogen signaling are of increasing concern, there is only limited information about their ability to interact with estrogen-binding proteins (SHBG) or receptors (ER). Recombinant ER?? and ?? were obtained after transient transfection of COS-7 cells with channel catfish ER cDNA. Plasma from adult female channel catfish was the source of SHBG. Tritiated estradiol ( 3H-E2) was used in standard radioligand-binding assays to characterize the binding properties of channel catfish SHBG (ccfSHBG) and to estimate the inhibition constants for various estrogenic compounds. Binding of 3H-E2 to ccfSHBG was saturable and of high affinity with a Kd (??SE) of 1.9??0.14nM and a Bmax of 14.3??2.4pmol/mg protein (n=3 assays). Additionally, ccfSHBG displayed binding specificity for androgens and estrogens. Endosulfan, 4-nonylphenol, and 4-octylphenol displaced 3H-E2 binding to ccfSHBG albeit only at very high concentrations, whereas dieldrin and atrazine showed little displacement activity even at the highest concentrations used. The synthetic estrogen ethynylestradiol had higher affinity than E2 for ccfSHBG. This finding differs from results with human and rainbow trout SHBG. The alkylphenolic compounds (4-octylphenol and 4-nonylphenol) displayed some ability to displace 3H-E2 binding from ER?? and ?? at high concentrations, but dieldrin and atrazine had little binding activity for both ER subtypes and endosulfan for ER??. The xenobiotics tested generally showed equivalent or greater affinity for ER?? than ER??, whereas natural estrogens had much greater affinity for ER?? than ER??. These observations suggest that results of studies using fish tissue ER extracts must be interpreted with caution, since both ER subtypes may be present, and that the binding of xenoestrogens to SHBG must be taken into account for proper assessment of endocrine

  5. A computational analysis of non-genomic plasma membrane progestin binding proteins: signaling through ion channel-linked cell surface receptors.

    Science.gov (United States)

    Morrill, Gene A; Kostellow, Adele B; Gupta, Raj K

    2013-12-11

    A number of plasma membrane progestin receptors linked to non-genomic events have been identified. These include: (1) α1-subunit of the Na(+)/K(+)-ATPase (ATP1A1), (2) progestin binding PAQR proteins, (3) membrane progestin receptor alpha (mPRα), (4) progesterone receptor MAPR proteins and (5) the association of nuclear receptor (PRB) with the plasma membrane. This study compares: the pore-lining regions (ion channels), transmembrane (TM) helices, caveolin binding (CB) motifs and leucine-rich repeats (LRRs) of putative progesterone receptors. ATP1A1 contains 10 TM helices (TM-2, 4, 5, 6 and 8 are pores) and 4 CB motifs; whereas PAQR5, PAQR6, PAQR7, PAQRB8 and fish mPRα each contain 8 TM helices (TM-3 is a pore) and 2-4 CB motifs. MAPR proteins contain a single TM helix but lack pore-lining regions and CB motifs. PRB contains one or more TM helices in the steroid binding region, one of which is a pore. ATP1A1, PAQR5/7/8, mPRα, and MAPR-1 contain highly conserved leucine-rich repeats (LRR, common to plant membrane proteins) that are ligand binding sites for ouabain-like steroids associated with LRR kinases. LRR domains are within or overlap TM helices predicted to be ion channels (pore-lining regions), with the variable LRR sequence either at the C-terminus (PAQR and MAPR-1) or within an external loop (ATP1A1). Since ouabain-like steroids are produced by animal cells, our findings suggest that ATP1A1, PAQR5/7/8 and mPRα represent ion channel-linked receptors that respond physiologically to ouabain-like steroids (not progestin) similar to those known to regulate developmental and defense-related processes in plants.

  6. Preclinical pharmacokinetics, tissue distribution and plasma protein binding of sodium (±-5-bromo-2-(α-hydroxypentyl benzoate (BZP, an innovative potent anti-ischemic stroke agent

    Directory of Open Access Journals (Sweden)

    Xin Tian

    2016-08-01

    Full Text Available Sodium (±-5-bromo-2-(α-hydroxypentyl benzoate (BZP is a potential cardiovascular drug and exerts potent neuroprotective effect against transient and long-term ischemic stroke in rats. BZP could convert into 3-butyl-6-bromo-1(3H-isobenzofuranone (Br-NBP in vitro and in vivo. However, the pharmacokinetic profiles of BZP and Br-NBP still have not been evaluated. For the purpose of investigating the pharmacokinetic profiles, tissue distribution and plasma protein binding of BZP and Br-NBP, a rapid, sensitive and specific method based on liquid chromatography coupled to mass spectrometry (LC-MS/MS has been developed for determination of BZP and Br-NBP in biological samples. The results indicated that BZP and Br-NBP showed a short elimination half-life, and pharmacokinetic profile in rats (3, 6 and 12 mg/kg; i.v. and beagle dogs (1, 2 and 4 mg/kg; i.v.gtt were obtained after single dosing of BZP. After multiple dosing of BZP, there was no significant accumulation of BZP and Br-NBP in the plasma of rats and beagle dogs. Following i.v. single dose (6 mg/kg to rats, BZP and Br-NBP were distributed rapidly into all tissues examined, with the highest concentrations of BZP and Br-NBP in lung and kidney, respectively. The brain distribution of Br-NBP in middle cerebral artery occlusion (MCAO rats was more than in normal rats (P<0.05. The plasma protein binding degree of BZP at three concentrations (8000, 20000 and 80000 ng/mL from rat, beagle dog and human plasma were 98.1~98.7%, 88.9~92.7% and 74.8%~83.7% respectively. In conclusion, both BZP and Br-NBP showed short half-life, good dose-linear pharmacokinetic profile, wide tissue distribution and different degree protein binding to various species plasma. This was the first preclinical pharmacokinetic investigation of BZP and Br-NBP in both rats and beagle dogs, which provided vital guidance for further preclinical research and the subsequent clinical trials.

  7. Toxicity challenges in environmental chemicals: Prediction of human plasma protein binding through quantitative structure-activity relationship (QSAR) models

    Science.gov (United States)

    The present study explores the merit of utilizing available pharmaceutical data to construct a quantitative structure-activity relationship (QSAR) for prediction of the fraction of a chemical unbound to plasma protein (Fub) in environmentally relevant compounds. Independent model...

  8. High performance liquid chromatographic determination of ibuprofen concentration and the binding rate of ibuprofen to human plasma proteins%高效液相色谱法测定布洛芬与人血浆蛋白的结合率

    Institute of Scientific and Technical Information of China (English)

    孙铮; 于治国; 毕开顺; 刘杰; 赵云丽; 陈晓辉

    2011-01-01

    目的:建立人血浆中布洛芬浓度的测定方法,测定布洛芬在人血浆中的蛋白结合率,并计算相关参数.方法:用平衡透析法以高效液相色谱法为检测手段测定血浆蛋白结合率.结果:布洛芬与人标准血浆的蛋白结合率为(95.4±0.3)%.结论:布洛芬与人血浆蛋白为高强度结合.%OBJECTIVE To develop a high performance liquid chromatographic(HPLC) method to determine the concentration of ibuprofen in human plasma,and study the binding rate of ibuprofen to plasma proteins and to calculate the binding parameters of ibuprofen to plasma proteins.METHODS The binding rate of ibuprofen to human plasma proteins was determined by equilibrium dialysis method and HPLC method.RFSULTS The binding rate of ibuprofen to human plasma proteins was (95.4 ± 0.3) %.CONCLUSION The binding rate of ibuprofen to human plasma proteins is high strength.

  9. Retinol Binding Protein 4 in children with Inflammatory Bowel Disease: a negative correlation with the disease activity.

    Science.gov (United States)

    Roma, E; Krini, M; Hantzi, E; Sakka, S; Panayiotou, I; Margeli, A; Papassotiriou, I; Kanaka-Gantenbein, C

    2012-10-01

    Retinol Binding Protein-4 (RBP-4), the action of which was initially thought to be only the transport of vitamin A, is a major circulating adipocytokine involved in the inflammation. We evaluated the serum RBP-4 levels in children with inflammatory bowel disease (IBD) and correlated them with transthyretin (TTR), inflammation markers, disease activity, and body mass index (BMI). In 41 children of mean age 11.9 ± 3.6 years (range 5-17.7 y) with IBD (19 with Crohn's disease (CD) and 22 with Ulcerative colitis (UC) serum RBP-4, TTR, Amyloid A (SAA), C-Reactive Protein (CRP), Erythrocyte Sedimentation Rate (ESR), disease activity and BMI were prospectively determined and compared with those of 42 matched controls. No difference in the RBP-4 and TTR serum levels, between patients and controls as well as between active and remission state of the disease was noticed. A negative correlation of serum RBP-4 with the disease activity, SAA and ESR and a positive correlation with TTR was found, but no significant correlation with CRP or BMI was found. Inflammation markers were significantly increased in patients compared to controls and had a positive correlation with the disease activity. RBP-4 negatively correlated with disease activity of children with IBD probably indicating a protective anti-inflammatory mechanism of action in addition to transport of vitamin A.

  10. Binding of alphaherpesvirus glycoprotein H to surface α4β1-integrins activates calcium-signaling pathways and induces phosphatidylserine exposure on the plasma membrane.

    Science.gov (United States)

    Azab, Walid; Gramatica, Andrea; Herrmann, Andreas; Osterrieder, Nikolaus

    2015-10-20

    Intracellular signaling connected to integrin activation is known to induce cytoplasmic Ca(2+) release, which in turn mediates a number of downstream signals. The cellular entry pathways of two closely related alphaherpesviruses, equine herpesviruses 1 and 4 (EHV-1 and EHV-4), are differentially regulated with respect to the requirement of interaction of glycoprotein H (gH) with α4β1-integrins. We show here that binding of EHV-1, but not EHV-4, to target cells resulted in a rapid and significant increase in cytosolic Ca(2+) levels. EHV-1 expressing EHV-4 gH (gH4) in lieu of authentic gH1 failed to induce Ca(2+) release, while EHV-4 with gH1 triggered significant Ca(2+) release. Blocking the interaction between gH1 and α4β1-integrins, inhibiting phospholipase C (PLC) activation, or blocking binding of inositol 1,4,5-triphosphate (IP3) to its receptor on the endoplasmic reticulum (ER) abrogated Ca(2+) release. Interestingly, phosphatidylserine (PS) was exposed on the plasma membrane in response to cytosolic calcium increase after EHV-1 binding through a scramblase-dependent mechanism. Inhibition of both Ca(2+) release from the ER and scramblase activation blocked PS scrambling and redirected virus entry to the endocytic pathway, indicating that PS may play a role in facilitating virus entry directly at the plasma membrane. Herpesviruses are a large family of enveloped viruses that infect a wide range of hosts, causing a variety of diseases. These viruses have developed a number of strategies for successful entry into different cell types. We and others have shown that alphaherpesviruses, including EHV-1 and herpes simplex virus 1 (HSV-1), can route their entry pathway and do so by manipulation of cell signaling cascades to ensure viral genome delivery to nuclei. We show here that the interaction between EHV-1 gH and cellular α4β1-integrins is necessary to induce emptying of ER calcium stores, which induces phosphatidylserine exposure on the plasma membrane

  11. Critical role of plasma corticosteroid-binding-globulin during stress to promote glucocorticoid delivery to the brain: impact on memory retrieval.

    Science.gov (United States)

    Minni, Amandine M; Dorey, Rodolphe; Piérard, Christophe; Dominguez, Gaëlle; Helbling, Jean-Christophe; Foury, Aline; Béracochéa, Daniel; Moisan, Marie-Pierre

    2012-10-01

    We aimed at demonstrating that corticosteroid binding globulin (CBG), a plasma glycoprotein binding glucocorticoids with high affinity in blood, endorses a major role under stress conditions by regulating free glucocorticoid access to the brain and thereby influences glucocorticoid-dependent behaviors. Hence, we compared CBG-deficient mice (Cbg-/-) and their controls (Cbg+/+) in a specific memory task, i.e. the delayed alternation behavior, requiring memory retrieval both under stress and nonstress conditions and previously shown to be dependent on hippocampal glucocorticoid levels. Our results evidence that Cbg-/- mice, unlike controls, remain insensitive to stress applied before memory retrieval. Furthermore, under stress conditions, we observed a blunted surge of corticosterone (CORT) in plasma and no free CORT rise in the hippocampus of Cbg-/-. Moreover, intrahippocampal infusion of CORT through implanted cannulae was used to mimic stress CORT rise before memory retrieval. This infusion of CORT reproduced memory retrieval impairments in Cbg-/- as in Cbg+/+ controls. Finally, we provide evidence that Cbg-/- mice exhibit a normal adrenal response to stress and ACTH. Given that CBG deficiency is known to markedly impact on CORT clearance from plasma, our current article demonstrates that Cbg-/- insensitivity in memory retrieval after stress results from the blunted CORT response due to increased CORT clearance. Overall, our data suggest that the impact of CBG genetic deficiency on various behavioral patterns reported previously stems from a smaller CORT reservoir in blood. Inasmuch as CBG discloses interindividual variations, such a parameter ought to be taken into account when studying stress-induced glucocorticoid action in brain.

  12. Seminal Plasma Characteristics and Expression of ATP-binding Cassette Transporter A1 (ABCA1) in Canine Spermatozoa from Ejaculates with Good and Bad Freezability.

    Science.gov (United States)

    Schäfer-Somi, S; Palme, N

    2016-04-01

    The composition of seminal plasma and the localization of the ATP-binding cassette transporter A1 (ABCA1) in spermatozoa from good and bad freezers were compared to frozen-thawed spermatozoa from the same dog. Ejaculates were obtained from 31 stud dogs, and the sperm-rich fraction (SRF) was kept for analysis. One aliquot was used for the analysis of concentration, progressive motility (P; CASA), viability (V; CASA) and leucocyte count, and the analysis was performed by flow cytometry (FITC-PNA/PI), SCSA and HOST. In seminal plasma, concentration of albumin, cholesterol, calcium, inorganic phosphate, sodium, potassium, zinc and copper was measured. Semen smears were prepared and evaluated for the expression of ABCA1. The remainder of each ejaculate was frozen. After thawing, the quality assessment was repeated and further smears were prepared. According to post-thaw semen quality, dogs were assigned to good freezers (n = 20) or bad freezers (n = 11), the latter were defined as 40% morphologically abnormal sperm and/or Bad freezers were older than good freezers (5.3 vs 3.4 years, p bad freezers, the percentage of sperm with ABCA1 signal in the acrosome was lower (26.3% vs 35.7%, p good freezers (p bad freezer sperm, an increase in acrosome damages coincided with an increased loss of cholesterol transporters and cell death, and a lower cholesterol concentration in seminal plasma. Follow-up studies revealed whether a relation exists between these findings.

  13. Sterol regulatory element-binding protein-1 determines plasma remnant lipoproteins and accelerates atherosclerosis in low-density lipoprotein receptor-deficient mice.

    Science.gov (United States)

    Karasawa, Tadayoshi; Takahashi, Akimitsu; Saito, Ryo; Sekiya, Motohiro; Igarashi, Masaki; Iwasaki, Hitoshi; Miyahara, Shoko; Koyasu, Saori; Nakagawa, Yoshimi; Ishii, Kiyoaki; Matsuzaka, Takashi; Kobayashi, Kazuto; Yahagi, Naoya; Takekoshi, Kazuhiro; Sone, Hirohito; Yatoh, Shigeru; Suzuki, Hiroaki; Yamada, Nobuhiro; Shimano, Hitoshi

    2011-08-01

    Sterol regulatory element-binding protein-1 (SREBP-1) is nutritionally regulated and is known to be a key transcription factor regulating lipogenic enzymes. The goal of this study was to evaluate the roles of SREBP-1 in dyslipidemia and atherosclerosis. Transgenic mice that overexpress SREBP-1c in the liver and SREBP-1-deficient mice were crossed with low-density lipoprotein receptor (LDLR)-deficient mice, and the plasma lipids and atherosclerosis were analyzed. Hepatic SREBP-1c overexpression in LDLR-deficient mice caused postprandial hypertriglyceridemia, increased very-low-density lipoprotein (VLDL) cholesterol, and decreased high-density lipoprotein cholesterol in plasma, which resulted in accelerated aortic atheroma formation. Conversely, absence of SREBP-1 suppressed Western diet-induced hyperlipidemia in LDLR-deficient mice and ameliorated atherosclerosis. In contrast, bone marrow-specific SREBP-1 deficiency did not alter the development of atherosclerosis. The size of nascent VLDL particles secreted from the liver was increased in SREBP-1c transgenic mice and reduced in SREBP-1-deficient mice, accompanied by upregulation and downregulation of phospholipid transfer protein expression, respectively. Hepatic SREBP-1c determines plasma triglycerides and remnant cholesterol and contributes to atherosclerosis in hyperlipidemic states. Hepatic SREBP-1c also regulates the size of nascent VLDL particles.

  14. Identification of 4-hydroxyheptachlorostyrene in polar bear plasma and its binding affinity to transthyretin : a metabolite of octachlorostyrene?

    NARCIS (Netherlands)

    Standau, C.D.; Meerts, I.A.T.M.; Letcher, R.J.; McAlees, A.J.; Chittim, B.; Brouwer, A.; Norstrom, R.J.

    2000-01-01

    A new compound, 4-hydroxyheptachlorostyrene (4-OH-HpCS), was identified as a major component in the chlorinated phenolic compound fraction of polar bear plasma. The structure was hypothesized to be 4-OH-HpCS based on mass spectral interpretation, the assumption that it was a metabolite of octachloro

  15. Identification of 4-hydroxyheptachlorostyrene in polar bear plasma and its binding affinity to transthyretin : a metabolite of octachlorostyrene?

    NARCIS (Netherlands)

    Standau, C.D.; Meerts, I.A.T.M.; Letcher, R.J.; McAlees, A.J.; Chittim, B.; Brouwer, A.; Norstrom, R.J.

    2000-01-01

    A new compound, 4-hydroxyheptachlorostyrene (4-OH-HpCS), was identified as a major component in the chlorinated phenolic compound fraction of polar bear plasma. The structure was hypothesized to be 4-OH-HpCS based on mass spectral interpretation, the assumption that it was a metabolite of

  16. Acute handling disturbance modulates plasma insulin-like growth factor binding proteins in rainbow trout (Oncorhynchus mykiss)

    Science.gov (United States)

    The effects of acute stressor exposure on proximal (growth hormone; GH) and distal (insulin-like growth factor-I; IGF-I and IGF-binding proteins) components of the somatotropic axis are poorly understood in finfish. We exposed rainbow trout (Oncorhynchus mykiss) to a 5-minute handling disturbance to...

  17. Association of Higher Plasma Vitamin D Binding Protein and Lower Free Calcitriol Levels with Tenofovir Disoproxil Fumarate Use and Plasma and Intracellular Tenofovir Pharmacokinetics: Cause of a Functional Vitamin D Deficiency?

    Science.gov (United States)

    Kiser, Jennifer J.; Stephensen, Charles B.; Hazra, Rohan; Flynn, Patricia M.; Wilson, Craig M.; Rutledge, Brandy; Bethel, James; Pan, Cynthia G.; Woodhouse, Leslie R.; Van Loan, Marta D.; Liu, Nancy; Lujan-Zilbermann, Jorge; Baker, Alyne; Kapogiannis, Bill G.; Gordon, Catherine M.

    2013-01-01

    Tenofovir disoproxil fumarate (TDF) causes bone, endocrine, and renal changes by an unknown mechanism(s). Data are limited on tenofovir pharmacokinetics and these effects. Using baseline data from a multicenter study of HIV-infected youth on stable treatment with regimens containing TDF (n = 118) or lacking TDF (n = 85), we measured cross-sectional associations of TDF use with markers of renal function, vitamin D-calcium-parathyroid hormone balance, phosphate metabolism (tubular reabsorption of phosphate and fibroblast growth factor 23 [FGF23]), and bone turnover. Pharmacokinetic-pharmacodynamic associations with plasma tenofovir and intracellular tenofovir diphosphate concentrations were explored among those receiving TDF. The mean age was 20.9 (standard deviation [SD], 2.0) years; 63% were male; and 52% were African American. Compared to the no-TDF group, the TDF group showed lower mean estimated glomerular filtration rates and tubular reabsorption of phosphate, as well as higher parathyroid hormone and 1,25-dihydroxy vitamin D [1,25-OH(2)D] levels. The highest quintile of plasma tenofovir concentrations was associated with higher vitamin D binding protein, lower free 1,25-OH(2)D, higher 25-OH vitamin D, and higher serum calcium. The highest quintile of intracellular tenofovir diphosphate concentration was associated with lower FGF23. Higher plasma tenofovir concentrations were associated with higher vitamin D binding protein and lower free 1,25-OH(2)D, suggesting a functional vitamin D deficiency explaining TDF-associated increased parathyroid hormone. The finding of lower FGF23 accompanying higher intracellular tenofovir diphosphate suggests that different mechanisms mediate TDF-associated changes in phosphate handling. Separate pharmacokinetic properties may be associated with distinct TDF toxicities: tenofovir with parathyroid hormone and altered calcium balance and tenofovir diphosphate with hypophosphatemia and FGF23 regulation. (The clinical trial

  18. [Can mannose-binding lectin and plasma level of soluble urokinase receptor be used in diagnosis and treatment monitorization of brucellosis patients?].

    Science.gov (United States)

    Karsen, Hasan; Cesur, Salih; Karaağaç, Leman; Binici, Irfan; Fidan, Yasemin; Oğüş, Elmas; Demiröz, Ali Pekcan

    2012-07-01

    The aim of this study was to evaluate the diagnostic value of serum mannose-binding lectin (MBL) and plasma soluble urokinase plasminogen activator receptor (SuPAR) levels in monitoring the treatment in patients with brucellosis, by comparing their levels before and after treatment with the values obtained from healthy control group. Thirty brucellosis patients (mean age: 25.8 ± 12.2 years; 15 were male) and 28 healthy controls (mean age: 29.3 ± 12.3 years; 15 were male) were included in the study. Patients were diagnosed with brucellosis according to the characteristic clinical findings and by brucella standard tube agglutination test (SAT) titer ≥ 1/160 and/or blood culture positivity. Serum MBL (Antibodyshop, Denmark) and plasma SuPAR (Virogates, Denmark) levels were investigated with commercial ELISA kits. In our study, no statistical significance was observed between the pre-treatment (13.8 ± 13.4 ng/ml) and post-treatment (12.4 ± 13.1 ng/ml) MBL levels of the patient group and MBL levels of the control group (16.5 ± 14.8 ng/ml) (p> 0.05). Moreover, the mean SuPAR levels measured in pre-treatment and post-treatment plasma samples of the brucellosis patients was 5.1 ± 1.9 ng/ml and 2.9 ± 1.3 ng/ml, respectively, while the mean SuPAR level was 1.8 ± 0.5 ng/ml in the control group. The difference between mean SuPAR levels of patients in pre- and post-treatment samples was found statistically significant (p 0.001). In conclusion, plasma SuPAR level would be a useful marker for the diagnosis and treatment follow up of the patients with brucellosis.

  19. MicroRNA-144 regulates hepatic ATP binding cassette transporter A1 and plasma high-density lipoprotein after activation of the nuclear receptor farnesoid X receptor.

    Science.gov (United States)

    de Aguiar Vallim, Thomas Q; Tarling, Elizabeth J; Kim, Tammy; Civelek, Mete; Baldán, Ángel; Esau, Christine; Edwards, Peter A

    2013-06-07

    The bile acid receptor farnesoid X receptor (FXR) regulates many aspects of lipid metabolism by variouscomplex and incompletely understood molecular mechanisms. We set out to investigate the molecular mechanisms for FXR-dependent regulation of lipid and lipoprotein metabolism. To identify FXR-regulated microRNAs that were subsequently involved in regulating lipid metabolism. ATP binding cassette transporter A1 (ABCA1) is a major determinant of plasma high-density lipoprotein (HDL)-cholesterol levels. Here, we show that activation of the nuclear receptor FXR in vivo increases hepatic levels of miR-144, which in turn lowers hepatic ABCA1 and plasma HDL levels. We identified 2 complementary sequences to miR-144 in the 3' untranslated region of ABCA1 mRNA that are necessary for miR-144-dependent regulation. Overexpression of miR-144 in vitro decreased both cellular ABCA1 protein and cholesterol efflux to lipid-poor apolipoprotein A-I protein, whereas overexpression in vivo reduced hepatic ABCA1 protein and plasma HDL-cholesterol. Conversely, silencing miR-144 in mice increased hepatic ABCA1 protein and HDL-cholesterol. In addition, we used tissue-specific FXR-deficient mice to show that induction of miR-144 and FXR-dependent hypolipidemia requires hepatic, but not intestinal, FXR. Finally, we identified functional FXR response elements upstream of the miR-144 locus, consistent with direct FXR regulation. We have identified a novel pathway involving FXR, miR-144, and ABCA1 that together regulate plasma HDL-cholesterol.

  20. Effect of seminal plasma zinc–binding proteins on motility and membrane integrity of canine spermatozoa stored at 5°C

    Directory of Open Access Journals (Sweden)

    Mogielnicka-Brzozowska Marzena

    2014-03-01

    Full Text Available Sperm surface binding sites for non-zinc-binding proteins (nZnBPs and zinc-binding proteins (ZnBPs were studied by the fluorescence technique with biotin-labelled proteins. The nZnBPs binding pattern was unspecific, no characteristic sites on plasmalemma were found. ZnBPs were attached mainly to the acrosomal region of sperm head and to the sperm flagellum. ZnBPs added to the incubation mixture of the canine spermatozoa allowed the preservation of higher values for total motility, progressive motility, curvilinear line velocity, straight line velocity, and beat cross frequency (P < 0.05, both at time 0 and after 1 h incubation at 5ºC. The addition of nZnBPs to the incubation mixture caused only weak positive effects when compared with control sample (PBS. A higher percentage of canine-ejaculated spermatozoa with intact membranes were observed when ejaculate was incubated with ZnBPs in comparison to control sample stored with PBS (P < 0.05 or nZnBPs (P < 0.05. Spermatozoa diluted with ZnBPs and nZnBPs exhibited a higher percentage of cells with active mitochondria when compared with control, both at time 0 and after 1 h; however, no statistical differences were observed. Our results emphasise the role of seminal plasma protein in securing the correct quantity and availability of zinc ions as a component regulating the motility of canine spermatozoa. The protective effect of ZnBPs against the cooling effect may be due to their ability of preventing sperm membrane damage.

  1. Binding of human coronary artery endothelial cells to plasma-treated titanium dioxide nanotubes of different diameters.

    Science.gov (United States)

    Flašker, Ajda; Kulkarni, Mukta; Mrak-Poljšak, Katjuša; Junkar, Ita; Čučnik, Saša; Žigon, Polona; Mazare, Anca; Schmuki, Patrik; Iglič, Aleš; Sodin-Semrl, Snezna

    2016-05-01

    Nanoscale topography in improving vascular response in vitro was established previously on various titanium surfaces. In the present study different surface nanotopographies that is different diameters of titanium dioxide (TiO2 ) nanotubes (NTs) were fabricated by electrochemical anodization and conditioned with highly reactive gaseous oxygen plasma. The morphology of different diameter NTs was studied by scanning electron microscopy and atomic force microscopy, while changes in chemical composition on the surface before and after plasma treatment were determined by X-ray photoelectron spectroscopy. Performance of human coronary artery endothelial cells (HCAEC) on those conditioned surfaces was studied in regard to cell proliferation, released IL-6 protein and immunofluorescence microscopy (IFM). We show that HCAEC function is dependent on the diameter of the TiO2 NTs, functioning far less optimally when bound to 100 nm TiO2 NTs as compared to Ti foil, 15 nm NTs or 50 nm NTs. There were improved, morphological cell shape changes, observed with IFM, between HCAEC growing on oxygen-rich plasma-treated versus nontreated 100 nm NTs. These endothelialized conditioned Ti nanosurfaces could elucidate optimization conditions necessary for vascular implants in coronary arteries.

  2. Cationic Au Nanoparticle Binding with Plasma Membrane-like Lipid Bilayers: Potential Mechanism for Spontaneous Permeation to Cells Revealed by Atomistic Simulations

    DEFF Research Database (Denmark)

    Heikkila, E.; Martinez-Seara, H.; Gurtovenko, A. A.

    2014-01-01

    Despite being chemically inert as a bulk material, nanoscale gold can pose harmful side effects to living organisms. In particular, cationic Au nanoparticles (AuNP+) of 2 nm diameter or less permeate readily through plasma membranes and induce cell death. We report atomistic simulations of cationic...... Au nanoparticles interacting with realistic membranes and explicit solvent using a model system that comprises two cellular compartments, extracellular and cytosolic, divided by two asymmetric lipid bilayers. The membrane-AuNP+ binding and membrane reorganization processes are discovered...... to be governed by cooperative effects where AuNP+, counterions, water, and the two membrane leaflets all contribute. On the extracellular side, we find that the nanoparticle has to cross a free energy barrier of about 5 k(B)T prior forming a stable contact with the membrane. This results in a rearrangement...

  3. Plasma levels of galectin-3-binding protein reflect type I interferon activity and are increased in patients with systemic lupus erythematosus

    DEFF Research Database (Denmark)

    Nielsen, Christoffer T; Lood, Christian; Ostergaard, Ole

    2014-01-01

    OBJECTIVE: Simple measures of type I interferon (IFN) activity constitute highly attractive biomarkers in systemic lupus erythematosus (SLE). We explore galectin-3-binding protein (G3BP) as a novel measure of type I IFN activity and serum/plasma biomarker in large independent cohorts of patients......-Denmark (DK) (n=70) and SLE-Sweden (SE) (n=68) cohorts were compared with the HC-DK (n=47) and HC-SE (n=50) cohorts and patients with systemic sclerosis (n=111). In 15 patients with SLE, serum G3BP in consecutive samples was correlated to disease activity. Correlation analysis between G3BP, clinical......-DK and SLE-SE cohorts compared with HCs and patients with systemic sclerosis (p

  4. Development of a formula for estimating plasma free cortisol concentration from a measured total cortisol concentration when elastase-cleaved and intact corticosteroid binding globulin coexist.

    Science.gov (United States)

    Nguyen, Phuong T T; Lewis, John G; Sneyd, James; Lee, Rita S F; Torpy, David J; Shorten, Paul R

    2014-05-01

    Cortisol bound to corticosteroid binding globulin (CBG) contributes up to 90% of the total cortisol concentration in circulation. Therefore, changes in the binding kinetics of cortisol to CBG can potentially impact on the concentration of free cortisol, the only form that is responsible for the physiological function of the hormone. When CBG is cleaved into elastase-cleaved CBG (eCBG) by the activity of neutrophil elastase, its affinity for cortisol is reduced. Therefore, when eCBG coexists with intact CBG (iCBG) in plasma, the calculation of free cortisol concentration based on the formulae that considers only one CBG pool with the same affinity for cortisol may be inappropriate. In this study, we developed in vivo and in vitro models of cortisol partitioning which considers two CBG pools, iCBG and eCBG, with different affinities for cortisol, and deduce a new formula for calculating plasma free cortisol concentration. The formula provides better estimates of free cortisol concentration than previously used formulae when measurements of the concentrations of the two CBG forms are available. The model can also be used to estimate the affinity of CBG and albumin for cortisol in different clinical groups. We found no significant difference in the estimated affinity of CBG and albumin for cortisol in normal, sepsis and septic shock groups, although free cortisol was higher in sepsis and septic shock groups. The in vivo model also demonstrated that the concentration of interstitial free cortisol is increased locally at a site of inflammation where iCBG is cleaved to form eCBG by the activity of elastase released by neutrophils. This supports the argument that the cleavage of iCBG at sites of inflammation leads to more lower-affinity eCBG and may be a mechanism that permits the local concentration of free cortisol to increase at these sites, while allowing basal free cortisol concentrations at other sites to remain unaffected.

  5. Binding of adrenergic ligands to liver plasma membrane preparations from the axolotl, Ambystoma mexicanum; the toad, Xenopus laevis; and the Australian lungfish, Neoceratodus forsteri.

    Science.gov (United States)

    Janssens, P A; Grigg, J A

    1988-09-01

    The beta-adrenergic ligand iodocyanopindolol (ICP) bound specifically to hepatic plasma membrane preparations from the axolotl, Ambystoma mexicanum (Bmax, 40 fmol/mg protein (P) at free concentration above 140 pM; KD, 42 pM); the toad, Xenopus laevis (Bmax, 200 fmol/mg P at 1 nM; KD, 300 pM); and the Australian lungfish, Neoceratodus forsteri (Bmax, 100 fmol/mg P at 5 nM). For the lungfish, the Scatchard plot was curved showing two classes of binding site with KD's of 20 and 500 pM. Neither the alpha 1-adrenergic ligand prazosin nor the alpha 2-adrenergic ligand yohimbine bound specifically to hepatic membrane preparations from any of the three species. Several adrenergic ligands displaced ICP from hepatic membrane preparations of all three species with KD's of Axolotl--propranolol, 50 nM; isoprenaline, 600 nM; adrenaline, 10 microM; phenylephrine, 20 microM; noradrenaline, 40 microM; and phentolamine, greater than 100 microM; X. laevis--propranolol, 30 nM; isoprenaline, 100 microM; adrenaline, 200 microM; noradrenaline, 300 microM; phenylephrine, 1 mM; and phentolamine, greater than 1 mM; N. forsteri,--propranolol, 25 nM; isoprenaline, 1 microM; adrenaline, 20 microM; phenylephrine, 35 microM; noradrenaline, 600 microM; and phentolamine, 400 microM. These findings suggest that alpha-adrenergic receptors are not present in hepatic plasma membrane preparations from these three species and that the hepatic actions of catecholamines are mediated via beta-adrenergic receptors. The order of binding of the beta-adrenergic ligands suggests that the receptors are of the beta 2 type.

  6. TNF and LT binding capacities in the plasma of arthritis patients: effect of etanercept treatment in juvenile idiopathic arthritis

    DEFF Research Database (Denmark)

    Gudbrandsdottir, S; Larsen, R; Sørensen, L K;

    2004-01-01

    Etanercept (Enbrel) induces a rapid and sustained decline in disease activity in the majority of patients with refractory juvenile idiopathic arthritis (JIA). For unknown reasons, however, a number of JIA patients fail to respond to this therapy. During this treatment neutralisation of tumour...... necrosis factor (TNF, previously termed TNF alpha) and lymphotoxin (LT, previously termed TNF beta) may be mediated by etanercept itself as well as by naturally occurring soluble TNF receptors. In light of this, it was of interest to study the total TNF neutralizing capacity in plasma before and during...... treatment with etanercept....

  7. Corrective Action Decision Document/Corrective Action Plan for Corrective Action Unit 413: Clean Slate II Plutonium Dispersion (TTR) Tonopah Test Range, Nevada. Revision 0

    Energy Technology Data Exchange (ETDEWEB)

    Matthews, Patrick [Navarro, Las Vegas, NV (United States)

    2017-05-01

    This Corrective Action Decision Document/Corrective Action Plan provides the rationale and supporting information for the selection and implementation of corrective actions at Corrective Action Unit (CAU) 413, Clean Slate II Plutonium Dispersion (TTR). CAU 413 is located on the Tonopah Test Range and includes one corrective action site, TA-23-02CS. CAU 413 consists of the release of radionuclides to the surface and shallow subsurface from the Clean Slate II (CSII) storage–transportation test conducted on May 31, 1963. The CSII test was a non-nuclear detonation of a nuclear device located inside a concrete bunker covered with 2 feet of soil. To facilitate site investigation and the evaluation of data quality objectives decisions, the releases at CAU 413 were divided into seven study groups: 1 Undisturbed Areas 2 Disturbed Areas 3 Sedimentation Areas 4 Former Staging Area 5 Buried Debris 6 Potential Source Material 7 Soil Mounds Corrective action investigation (CAI) activities, as set forth in the CAU 413 Corrective Action Investigation Plan, were performed from June 2015 through May 2016. Radionuclides detected in samples collected during the CAI were used to estimate total effective dose using the Construction Worker exposure scenario. Corrective action was required for areas where total effective dose exceeded, or was assumed to exceed, the radiological final action level (FAL) of 25 millirem per year. The results of the CAI and the assumptions made in the data quality objectives resulted in the following conclusions: The FAL is exceeded in surface soil in SG1, Undisturbed Areas; The FAL is assumed to be exceeded in SG5, Buried Debris, where contaminated debris and soil were buried after the CSII test; The FAL is not exceeded at SG2, SG3, SG4, SG6, or SG7. Because the FAL is exceeded at CAU 413, corrective action is required and corrective action alternatives (CAAs) must be evaluated. For CAU 413, three CAAs were evaluated: no further action, clean closure, and

  8. Mannose-binding lectin 2 (Mbl2 gene polymorphisms are related to protein plasma levels, but not to heart disease and infection by Chlamydia

    Directory of Open Access Journals (Sweden)

    M.A.F. Queiroz

    Full Text Available The presence of the single nucleotide polymorphisms in exon 1 of the mannose-binding lectin 2 (MBL2 gene was evaluated in a sample of 159 patients undergoing coronary artery bypass surgery (71 patients undergoing valve replacement surgery and 300 control subjects to investigate a possible association between polymorphisms and heart disease with Chlamydia infection. The identification of the alleles B and D was performed using real time polymerase chain reaction (PCR and of the allele C was accomplished through PCR assays followed by digestion with the restriction enzyme. The comparative analysis of allelic and genotypic frequencies between the three groups did not reveal any significant difference, even when related to previous Chlamydia infection. Variations in the MBL plasma levels were influenced by the presence of polymorphisms, being significantly higher in the group of cardiac patients, but without representing a risk for the disease. The results showed that despite MBL2 gene polymorphisms being associated with the protein plasma levels, the polymorphisms were not enough to predict the development of heart disease, regardless of infection with both species of Chlamydia.

  9. Reticulomics: Protein-Protein Interaction Studies with Two Plasmodesmata-Localized Reticulon Family Proteins Identify Binding Partners Enriched at Plasmodesmata, Endoplasmic Reticulum, and the Plasma Membrane.

    Science.gov (United States)

    Kriechbaumer, Verena; Botchway, Stanley W; Slade, Susan E; Knox, Kirsten; Frigerio, Lorenzo; Oparka, Karl; Hawes, Chris

    2015-11-01

    The endoplasmic reticulum (ER) is a ubiquitous organelle that plays roles in secretory protein production, folding, quality control, and lipid biosynthesis. The cortical ER in plants is pleomorphic and structured as a tubular network capable of morphing into flat cisternae, mainly at three-way junctions, and back to tubules. Plant reticulon family proteins (RTNLB) tubulate the ER by dimerization and oligomerization, creating localized ER membrane tensions that result in membrane curvature. Some RTNLB ER-shaping proteins are present in the plasmodesmata (PD) proteome and may contribute to the formation of the desmotubule, the axial ER-derived structure that traverses primary PD. Here, we investigate the binding partners of two PD-resident reticulon proteins, RTNLB3 and RTNLB6, that are located in primary PD at cytokinesis in tobacco (Nicotiana tabacum). Coimmunoprecipitation of green fluorescent protein-tagged RTNLB3 and RTNLB6 followed by mass spectrometry detected a high percentage of known PD-localized proteins as well as plasma membrane proteins with putative membrane-anchoring roles. Förster resonance energy transfer by fluorescence lifetime imaging microscopy assays revealed a highly significant interaction of the detected PD proteins with the bait RTNLB proteins. Our data suggest that RTNLB proteins, in addition to a role in ER modeling, may play important roles in linking the cortical ER to the plasma membrane.

  10. Mannose-binding lectin 2 (Mbl2) gene polymorphisms are related to protein plasma levels, but not to heart disease and infection by Chlamydia

    Science.gov (United States)

    Queiroz, M.A.F.; Gomes, S.T.M.; Almeida, N.C.C.; Souza, M.I.M.; Costa, S.R.C.F.; Hermes, R.B.; Lima, S.S.; Zaninotto, M.M.; Fossa, M.A.A.; Maneschy, M.A.; Martins-Feitosa, R.N.; Azevedo, V.N.; Machado, L.F.A.; Ishak, M.O.G.; Ishak, R.; Vallinoto, A.C.R.

    2016-01-01

    The presence of the single nucleotide polymorphisms in exon 1 of the mannose-binding lectin 2 (MBL2) gene was evaluated in a sample of 159 patients undergoing coronary artery bypass surgery (71 patients undergoing valve replacement surgery and 300 control subjects) to investigate a possible association between polymorphisms and heart disease with Chlamydia infection. The identification of the alleles B and D was performed using real time polymerase chain reaction (PCR) and of the allele C was accomplished through PCR assays followed by digestion with the restriction enzyme. The comparative analysis of allelic and genotypic frequencies between the three groups did not reveal any significant difference, even when related to previous Chlamydia infection. Variations in the MBL plasma levels were influenced by the presence of polymorphisms, being significantly higher in the group of cardiac patients, but without representing a risk for the disease. The results showed that despite MBL2 gene polymorphisms being associated with the protein plasma levels, the polymorphisms were not enough to predict the development of heart disease, regardless of infection with both species of Chlamydia. PMID:27982280

  11. Effect of the binding interaction of an emissive niacin derivative on the conformation and activity of a model plasma protein: A spectroscopic and simulation-based approach.

    Science.gov (United States)

    Sett, Riya; Ganguly, Aniruddha; Guchhait, Nikhil

    2016-11-01

    The present work demonstrates a detailed photophysics of bio-active drug-like acid viz., 2-hydroxynicotinic acid (2-HNA) and its interaction with a model plasma protein Bovine Serum Albumin (BSA). The drug which is in essence a vitamin-B3 derivative, is capable of exhibiting ultrafast lactim-lactam cross-over response and thereby the modulation of the lactam emission within the bio-environment of the protein has been depicted spectroscopically to reveal the drug-protein interaction. Apart from evaluating the binding constant, the probable location of the neutral drug molecule within the protein cavity (hydrophobic subdomain IIIA) has been explored by AutoDock-based blind docking simulation technique. In this microheterogeneous medium, slow solvent reorientation time with respect to the emissive lifetime of the drug explicate the Red Edge Effect (REE). To complement the findings about the binding process, chaotrope-induced protein denaturation has also been inspected. The probe also illustrates a perceptible difference in rotational relaxation time in confined medium than in aqueous medium which strengthen our verdict. Unfolding of the protein in the presence of the drug molecule has been probed by the decrease of the α-helical content, obtained via circular dichroism (CD) spectroscopy, which is also supported by the gradual slaughter of the esterase activity of the protein in the presence of the drug molecule.

  12. Binding of SEC11 indicates its role in SNARE recycling after vesicle fusion and identifies two pathways for vesicular traffic to the plasma membrane.

    Science.gov (United States)

    Karnik, Rucha; Zhang, Ben; Waghmare, Sakharam; Aderhold, Christin; Grefen, Christopher; Blatt, Michael R

    2015-03-01

    SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) proteins drive vesicle fusion in all eukaryotes and contribute to homeostasis, pathogen defense, cell expansion, and growth in plants. Two homologous SNAREs, SYP121 (=SYR1/PEN1) and SYP122, dominate secretory traffic to the Arabidopsis thaliana plasma membrane. Although these proteins overlap functionally, differences between SYP121 and SYP122 have surfaced, suggesting that they mark two discrete pathways for vesicular traffic. The SNAREs share primary cognate partners, which has made separating their respective control mechanisms difficult. Here, we show that the regulatory protein SEC11 (=KEULE) binds selectively with SYP121 to affect secretory traffic mediated by this SNARE. SEC11 rescued traffic block by dominant-negative (inhibitory) fragments of both SNAREs, but only in plants expressing the native SYP121. Traffic and its rescue were sensitive to mutations affecting SEC11 interaction with the N terminus of SYP121. Furthermore, the domain of SEC11 that bound the SYP121 N terminus was itself able to block secretory traffic in the wild type and syp122 but not in syp121 mutant Arabidopsis. Thus, SEC11 binds and selectively regulates secretory traffic mediated by SYP121 and is important for recycling of the SNARE and its cognate partners.

  13. Plasma-free fatty acids, fatty acid-binding protein 4, and mortality in older adults (from the Cardiovascular Health Study).

    Science.gov (United States)

    Miedema, Michael D; Maziarz, Marlena; Biggs, Mary L; Zieman, Susan J; Kizer, Jorge R; Ix, Joachim H; Mozaffarian, Dariush; Tracy, Russell P; Psaty, Bruce M; Siscovick, David S; Mukamal, Kenneth J; Djousse, Luc

    2014-09-15

    Plasma-free fatty acids (FFAs) are largely derived from adipose tissue. Elevated levels of FFA and fatty acid-binding protein 4 (FABP4), a key cytoplasmic chaperone of fatty acids, have been associated with adverse cardiovascular outcomes, but limited data are available on the relation of these biomarkers with cardiovascular and total mortality. We studied 4,707 participants with a mean age of 75 years who had plasma FFA and FABP4 measured in 1992 to 1993 as part of the Cardiovascular Health Study, an observational cohort of community-dwelling older adults. Over a median follow-up of 11.8 years, 3,555 participants died. Cox proportional hazard regression was used to determine the association between FFA, FABP4, and mortality. In fully adjusted models, FFA were associated with dose-dependent significantly higher total mortality (hazard ratio [HR] per SD: 1.14, 95% confidence interval [CI] 1.09 to 1.18), but FABP4 levels were not (HR 1.04, 95% CI 0.98 to 1.09). In a cause-specific mortality analysis, higher concentrations of FFA were associated with significantly higher risk of death because of cardiovascular disease, dementia, infection, and respiratory causes but not cancer or trauma. We did not find evidence of an interaction between FFA and FABP4 (p = 0.45), but FABP4 appeared to be associated with total mortality differentially in men and women (HR 1.17, 95% CI 1.08 to 1.26 for men; HR 1.02, 95% CI 0.96 to 1.07 for women, interaction p value <0.001). In conclusion, in a cohort of community-dwelling older subjects, elevated plasma concentrations of FFA, but not FABP4, were associated with cardiovascular and noncardiovascular mortality.

  14. Increase in skin autofluorescence and release of heart-type fatty acid binding protein in plasma predicts mortality of hemodialysis patients.

    Science.gov (United States)

    Arsov, Stefan; Trajceska, Lada; van Oeveren, Wim; Smit, Andries J; Dzekova, Pavlina; Stegmayr, Bernd; Sikole, Aleksandar; Rakhorst, Gerhard; Graaff, Reindert

    2013-07-01

    Advanced glycation end-products (AGEs) are uremic toxins that accumulate progressively in hemodialysis (HD) patients. The aim of this study was to assess the 1-year increase in skin autofluorescence (ΔAF), a measure of AGEs accumulation and plasma markers, as predictors of mortality in HD patients. One hundred sixty-nine HD patients were enrolled in this study. Skin autofluorescence was measured twice, 1 year apart using an AGE Reader (DiagnOptics Technologies BV, Groningen, The Netherlands). Besides routine blood chemistry, additional plasma markers including superoxide dismutase, myeloperoxydase, intercellular adhesion molecule 1 (ICAM-1), C-reactive protein (hs-CRP), heart-type fatty acid binding protein (H-FABP), and von Willebrand factor were measured at baseline. The mortality of HD patients was followed for 36 months. Skin autofluorescence values of the HD patients at the two time points were significantly higher (P < 0.001) than those of healthy subjects of the same age. Mean 1-year ΔAF of HD patients was 0.16 ± 0.06, which was around seven- to ninefold higher than 1-year ΔAF in healthy subjects. Multivariate Cox regression showed that age, hypertension, 1-year ΔAF, hs-CRP, ICAM-1, and H-FABP were independent predictors of overall mortality. Hypertension, 1-year ΔAF, hs-CRP, and H-FABP were also independent predictors of cardiovascular mortality. One-year ΔAF and plasma H-FABP, used separately and in combination, are strong predictors of overall and cardiovascular mortality in HD patients.

  15. Analysis of flurbiprofen, ketoprofen and etodolac enantiomers by pre-column derivatization RP-HPLC and application to drug-protein binding in human plasma.

    Science.gov (United States)

    Jin, Yin-Xiu; Tang, Yi-Hong; Zeng, Su

    2008-04-14

    A stereoselective reversed-phase high-performance liquid chromatography (HPLC) assay to determine the enantiomers of flurbiprofen, ketoprofen and etodolac in human plasma was developed. Chiral drug enantiomers were extracted from human plasma with liquid-liquid extraction. Then flurbiprofen and ketoprofen enantiomers reacted with the acylation reagent thionyl chloride and pre-column chiral derivatization reagent (S)-(-)-alpha-(1-naphthyl)ethylamine (S-NEA), and etodolac enantiomers reacted with S-NEA using 1-(3-dimethylaminopropyl)-3-ethyl-carbodiimide (EDC) and 1-hydroxybenzotriazole (HOBT) as coupling agents. The derivatized products were separated on an Agilent Zorbax C18 (4.6 mm x 250 mm, 5 microm) column with a mixture of acetonitrile-0.01 mol.L(-1) phosphate buffer (pH 4.5) (70:30, v/v) for flurbiprofen enantiomers, acetonitrile-0.01 mol.L(-1) phosphate buffer (pH 4.5) (60:40, v/v) for ketoprofen enantiomers and methonal-0.01 mol.L(-1) potassium dihydrogen phosphate buffer (pH 4.5) (88:12, v/v) for etodolac enantiomers as mobile phase. The flow of mobile phase was set at 0.8 mL.min(-1) and the detection wavelength of UV detector was set at 250 nm for flurbiprofen and ketoprofen enantiomers and 278 nm for etodolac enantiomers. The assay was linear from 0.5 to 50 microg.mL(-1) for each enantiomer. The inter- and intra-day precision (R.S.D.) was less than 10% and the average extraction recovery was more than 87% for each enantiomer. The limit of quantification for the method was 0.5 microg.mL(-1) (R.S.D.<10%, n=5). The method developed was used to study the drug-protein binding of flurbiprofen, ketoprofen and etodolac enantiomers in human plasma. The results showed that the stereoselective binding of etodolac enantiomer was observed and flurbiprofen and ketoprofen enantiomers were not.

  16. Independent associations of polymorphisms in vitamin D binding protein (GC) and vitamin D receptor (VDR) genes with obesity and plasma 25OHD3 levels demonstrate sex dimorphism.

    Science.gov (United States)

    Almesri, Norah; Das, Nagalla S; Ali, Muhallab E; Gumaa, Khalid; Giha, Hayder Ahmed

    2016-04-01

    We investigated a possible association between polymorphisms in vitamin D binding protein (GC) and vitamin D receptor (VDR) genes and obesity in Bahraini adults. For this purpose, 406 subjects with varying body mass indexes (BMIs) were selected. Plasma levels of 25-hydroxyvitamin D3 (25OHD3) were measured by chemiluminescence immunoassay. Six single nucleotide polymorphisms, 2 in the VDR gene (rs731236 TC and rs12721377 AG) and 4 in the GC gene (rs2282679 AC, rs4588 CA, rs7041 GT, and rs2298849 TC), were genotyped by real-time polymerase chain reaction. We found that the rs7041 minor allele (G) and rare genotype (GG) were associated with higher BMI (p = 0.007 and p = 0.012, respectively), but they did not influence 25OHD3 levels. However, the minor alleles of rs2282679 (A) and rs4588 (C) were associated with low 25OHD3 plasma levels (p = 0.039 and p = 0.021, respectively), but not with BMI. Having categorized the subjects based on their sex, we found that (i) rs7041 GG associated with high BMI in females (p = 0.003), (ii) rs4588 CC associated with high BMI in females (p = 0.034) and low 25OHD3 levels in males (p = 0.009), and (iii) rs12721377 AA associated with low 25OHD3 levels in females (p = 0.039). Notably, none of the common haplotypes (6 in the GC gene and 3 in the VDR gene) were associated with BMI. Therefore, polymorphisms in the GC (rs2282679, rs4588, rs7041) and VDR (rs12721377) genes were independently associated with obesity and 25OHD3 levels with a clear sex dimorphism.

  17. Low Abdominal NIRS Values and Elevated Plasma Intestinal Fatty Acid-Binding Protein in a Premature Piglet Model of Necrotizing Enterocolitis.

    Directory of Open Access Journals (Sweden)

    Irving J Zamora

    Full Text Available To identify early markers of necrotizing enterocolitis (NEC, we hypothesized that continuous abdominal near-infrared spectroscopy (A-NIRS measurement of splanchnic tissue oxygen saturation and intermittent plasma intestinal fatty-acid binding protein (pI-FABP measured every 6 hours can detect NEC prior to onset of clinical symptoms. Premature piglets received parenteral nutrition for 48-hours after delivery, followed by enteral feeds every three hours until death or euthanasia at 96-hours. Continuous A-NIRS, systemic oxygen saturation (SpO2, and heart rate were measured while monitoring for clinical signs of NEC. Blood samples obtained at 6-hour intervals were used to determine pI-FABP levels by ELISA. Piglets were classified as fulminant-NEC (f-NEC, non-fulminant-NEC (nf-NEC and No-NEC according to severity of clinical and histologic features. Of 38 piglets, 37% (n=14 developed nf-NEC, 18% (n=7 developed f-NEC and 45% (n=17 had No-NEC. There were significant differences in baseline heart rate (p=0.008, SpO2 (p0.25ng/mL identified animals with NEC (68% sensitivity and 90% specificity. NIRS is a real-time, non-invasive tool that can serve as a diagnostic modality for NEC. In premature piglets, low A-NIRS in the early neonatal period and increased variability during initial feeds are highly predictive of NEC, which is then confirmed by rising plasma I-FABP levels. These modalities may help identify neonates with NEC prior to clinical manifestations of disease.

  18. Evaluation of gel electrophoresis techniques and laser ablation-inductively coupled plasma-mass spectrometry for screening analysis of Zn and Cu-binding proteins in plankton.

    Science.gov (United States)

    Jiménez, Maria S; Rodriguez, L; Bertolin, Juan R; Gomez, Maria T; Castillo, Juan R

    2013-01-01

    The determination of metal-binding proteins in plankton is important because of their involvement in photosynthesis, which is fundamental to the biogeochemical cycle of the oceans and other ecosystems. We have elaborated a new strategy for screening of Cu and Zn-containing proteins in plankton on the basis of separation of proteins by use of Blue-Native PAGE (BN-PAGE), which entails use of a non-denaturing Tris-tricine system and detection of metals in the proteins by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). For comparison, denaturing PAGE based on Tris-glycine and Tris-tricine systems and Anodic-Native PAGE have also been investigated. A large number of protein bands with MW between 20 and 75 kDa were obtained by use of Tris-glycine PAGE but detection of metals by LA-ICP-MS was unsuccessful because of loss of metals from the proteins during the separation process. Different protein extraction, purification, and preconcentration methods were evaluated, focussing on both issues-achieving the best extraction and characterization of the proteins while maintaining the integrity of metal-protein binding in the plankton sample. Use of 25 mmol L(-1) Tris-HCl and a protease inhibitor as extraction buffer with subsequent ultrafiltration and acetone precipitation was the most efficient means of sample preparation. Two Cu and Zn proteins were detected, a protein band corresponding to a MW of 60 kDa and another poorly resolved band with a MW between 15 and 35 kDa.

  19. A Substructure Combination Strategy To Create Potent and Selective Transthyretin Kinetic Stabilizers That Prevent Amyloidogenesis and Cytotoxicity

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Sungwook; Reixach, Natlia; Connelly, Stephen; Johnson, Steven M.; Wilson, Ian A.; Kelly, Jeffery W. (Scripps)

    2010-08-13

    Transthyretin aggregation-associated proteotoxicity appears to cause several human amyloid diseases. Rate-limiting tetramer dissociation and monomer misfolding of transthyretin (TTR) occur before its aggregation into cross-{beta}-sheet amyloid fibrils. Small molecule binding to and preferential stabilization of the tetrameric state of TTR over the dissociative transition state raises the kinetic barrier for dissociation, imposing kinetic stabilization on TTR and preventing aggregation. This is an effective strategy to halt neurodegeneration associated with polyneuropathy, according to recent placebo-controlled clinical trial results. In three recent papers, we systematically ranked possibilities for the three substructures composing a typical TTR kinetic stabilizer, using fibril inhibition potency and plasma TTR binding selectivity data. Herein, we have successfully employed a substructure combination strategy to use these data to develop potent and selective TTR kinetic stabilizers that rescue cells from the cytotoxic effects of TTR amyloidogenesis. Of the 92 stilbene and dihydrostilbene analogues synthesized, nearly all potently inhibit TTR fibril formation. Seventeen of these exhibit a binding stoichiometry of >1.5 of a maximum of 2 to plasma TTR, while displaying minimal binding to the thyroid hormone receptor (<20%). Six analogues were definitively categorized as kinetic stabilizers by evaluating dissociation time-courses. High-resolution TTR-(kinetic stabilizer)2 crystal structures (1.31-1.70 {angstrom}) confirmed the anticipated binding orientation of the 3,5-dibromo-4-hydroxyphenyl substructure and revealed a strong preference of the isosteric 3,5-dibromo-4-aminophenyl substructure to bind to the inner thyroxine binding pocket of TTR.

  20. Insulin sensitivity is independent of lipid binding protein trafficking at the plasma membrane in human skeletal muscle: effect of a 3-day, high-fat diet.

    Science.gov (United States)

    Jordy, Andreas B; Serup, Annette K; Karstoft, Kristian; Pilegaard, Henriette; Kiens, Bente; Jeppesen, Jacob

    2014-11-01

    The aim of the present study was to investigate lipid-induced regulation of lipid binding proteins in human skeletal muscle and the impact hereof on insulin sensitivity. Eleven healthy male subjects underwent a 3-day hypercaloric and high-fat diet regime. Muscle biopsies were taken before and after the diet intervention, and giant sarcolemmal vesicles were prepared. The high-fat diet induced decreased insulin sensitivity, but this was not associated with a relocation of FAT/CD36 or FABPpm protein to the sarcolemma. However, FAT/CD36 and FABPpm mRNA, but not the proteins, were upregulated by increased fatty acid availability. This suggests a time dependency in the upregulation of FAT/CD36 and FABPpm protein during high availability of plasma fatty acids. Furthermore, we did not detect FATP1 and FATP4 protein in giant sarcolemmal vesicles obtained from human skeletal muscle. In conclusion, this study shows that a short-term lipid-load increases mRNA content of key lipid handling proteins in human muscle. However, decreased insulin sensitivity after a high-fat diet is not accompanied with relocation of FAT/CD36 or FABPpm protein to the sarcolemma. Finally, FATP1 and FATP4 protein was located intracellularly but not at the sarcolemma in humans. Copyright © 2014 the American Physiological Society.

  1. 超滤法测定丹酚酸A的血浆蛋白结合率%Determination of Plasma Protein Binding Rate of Salvianol Acid A by Ultrafiltration

    Institute of Scientific and Technical Information of China (English)

    陈豆; 涂星; 张英丰

    2013-01-01

    Objective: To establish a method for the determination of salvianolic acid A in the plasma samples, and then study the plasma protein binding rate of salvianolic acid A. This can provide a reference to the further study of salvianol acid A in its metabolism, pharmacokinetics and clinical treatment. Method: The ultrafiltration was employed to determine the plasma protein binding rate of salvianol acid A in BSA plasma samples, rat plasma samples, the New Zealand rabbit plasma samples and beagle dog plasma samples. The plasma concentrations of salvianol acid A were measured by RP-HPLC. Result: The calibration curve of salvianolic acid A was linear within the ranges of 0. 01 -2. 5 mg ·L-1 ( r = 0. 999 4) . The average plasma protein binding rates of salvianol acid A with BSA, rat plasma, the New Zealand rabbit plasma and beagle dog plasma were (99.79 ± 0. 02) % , (99. 79 ± 0. 03 ) % , (99. 73 ± 0. 06) % , (99. 81 ± 0. 03 ) % in the plasma concentrations of 5. 0, 50.0, 100.0 mg · L-1 respectively. Conclusion: The method has high sensitivity, good specificity and reproduction, with simple management thus fulfilling the requirement. Salvianol acid A shows a high binding power to plasma protein. , and this was independent of the investigated concentrations and the different species.%目的:建立血浆样品中丹酚酸A浓度的分析方法,并测定丹酚酸A的血浆蛋白结合率,为进一步展开丹酚酸A的代谢和药动学研究及指导临床用药提供参考.方法:采用超滤法和HPLC测定丹酚酸A在牛血清白蛋白(BSA),大鼠血浆,新西兰兔血浆和比格犬血浆中的蛋白结合率.结果:丹酚酸A在0.01~2.5 mg·L-1线性良好,标准曲线方程为Y =0.8073X+0.013 2(r =0.999 4).丹酚酸A与BSA,大鼠血浆,新西兰兔血浆和比格犬血浆在5.0,50.0,100.0 mg·L-1的含药血浆中其平均蛋白结合率分别为(99.79±0.02)%,(99.79±0.03)%,(99.73±0.06)%,(99.81±0.03)%.结论:所建立的方法灵敏度高,专属性

  2. Development of validated HPLC-UV method for simultaneous determination of Metformin, Amlodipine, Glibenclamide and Atorvastatin in human plasma and application to protein binding studies

    Directory of Open Access Journals (Sweden)

    Pawan K. Porwal

    2017-06-01

    Full Text Available A simple, sensitive, fast, and economical HPLC method was developed and validated for simultaneous estimation of two fixed dose combinations frequently prescribed in diabetes (Metformin plus Glibenclamide and hypertension with dyslipidemia (Amlodipine plus Atorvastatin in Human plasma for the first time. The validated HPLC method was used to quantify the concentration of selected actives in ultrafiltrate. Optimum separation conditions were obtained with Water’s Novapack Phenyl (150 mm × 4.6 mm, i.d., 5.0 μm column with mobile phase consisting of 0.1% Phosphoric acid (pH 3.0 and acetonitrile (ACN in gradient mode with column oven temperature maintained at 30 °C and elution monitored by a UV detector at 227 nm. Protein precipitation was employed to extract the selected analyte form human plasma. The recoveries were more than 90% for all analytes in cold aqueous 10% trichloroacetic acid (TCA and acetonitrile. The optimized HPLC-UV was validated in the calibration range of 10–10,000 ng mL−1 for Metformin, 25–5000 ng mL−1 for amlodipine, 50–10,000 ng mL−1 for glibenclamide and 10–5000 ng mL−1 for atorvastatin. The mean relative error was least when weighing of 1/×2 was applied for calibration curve. The accuracy of samples for six replicate measurements at LLOQ level was within limit. The precision and accuracy of samples for six replicate measurements at LLOQ level was within limit. The validated method was applied for quantitation of selected analytes in ultrafiltrate from protein binding experiments. A four to five fold increase in unbound fraction was observed when spiked to human serum albumin. Further the unbound fraction of highly albumin bound drugs was increased nearly to double when incubated with Gly-HSA as compare to HSA.

  3. Determination of human plasma protein binding rate of oridonin by ultrafiltration and high-performance liquid chromatography%超滤法测定冬凌草甲素的血浆蛋白结合率

    Institute of Scientific and Technical Information of China (English)

    陈莹; 蔡爽

    2013-01-01

    Objective To study the plasma protein binding rate of oridonin in human plasma,rat plasma and rabbit plasma to provide the basis for designing reasonable dosage regimen.Methods Ultraflitration method was used to imitate the binding rates of oridonin.High performance liquid chromatography (HPLC) was employed to determine the concentration of oridonin,and then on the basis of which protein binding rates were calculated.Results The plasma protein binding rate of oridonin at the concentration of 5.4,21.6 and 108.0 μg/mL with normal human plasma were 80.6%,78.3% and 76.2%,with rat plasma were 69.3%,67.5% and 68.1%,while with rabbit plasma were 65.2%,66.7% and 64.3%,respectively.Conclusion The ultraflitration-HPLC method is simple,rapid,reliable and can be usd to determine the drug concentration of oridonin properly.The results suggested that the protein binding rates of oridonin were of middle strength in binding to plasma protein of rabbits,rats and human and it is not proportionally dependent on plasma concentration of oridonin.%目的 测定冬凌草甲素在健康人血浆、大鼠血浆和家兔血浆中的蛋白结合率,为设计合理的给药方案提供依据.方法 采用超滤法结合高效液相色谱法(HPLC)对冬凌草甲素与大鼠、家兔和健康人血浆的蛋白结合率进行测定.结果 冬凌草甲素在覆盖临床给药范围的5.4,21.6和108.0 μg/mL下与健康人血浆的血浆蛋白结合率分别为80.6%,78.3%,76.2%,与大鼠的血浆蛋白结合率分别为69.3%,67.5%,68.1%,与家兔的血浆蛋白结合率分别为65.2%,66.7%,64.3%.覆盖临床给药浓度的3个浓度的冬凌草甲素在同种血浆中血浆蛋白结合率差异无统计学意义(P>0.05).结论 结果表明本法简便快速、灵敏度高、专属性好,能够满足测试生物样品的要求,超滤法结合高效液相色谱法可用于冬凌草甲素血浆蛋白结合率的检测.在本研究试验浓度范围内,

  4. Postprandial changes in plasma growth hormone, insulin, insulin-like growth factor (IGF)-I, and IGF-binding proteins in coho salmon fasted for varying periods.

    Science.gov (United States)

    Shimizu, Munetaka; Cooper, Kathleen A; Dickhoff, Walton W; Beckman, Brian R

    2009-08-01

    We examined postprandial changes in circulating growth hormone (GH), insulin, insulin-like growth factor (IGF)-I, and IGF-binding proteins (IGFBPs) in yearling coho salmon under different feeding regimes. Fish were initially fasted for 1 day, 1 wk, or 3 wk. Fasted fish were then fed, and blood was collected at 4-h intervals over 26 h. After the various periods of fasting, basal levels of insulin were relatively constant, whereas those of IGF-I, IGFBPs and GH changed in proportion to the duration of the fast. A single meal caused a rapid, large increase in the circulating insulin levels, but the degree of the increase was influenced by the fasting period. IGF-I showed a moderate increase 2 h after the meal but only in the regularly fed fish. Plasma levels of 41-kDa IGFBP were increased in all groups within 6 h after the single meal. The fasting period did not influence the response of 41-kDa IGFBP to the meal. IGFBP-1 and GH decreased after the meal to the same extent among groups regardless of the fasting period. The present study shows that insulin and IGF-I respond differently to long (weeks)- and short (hours)-term nutritional changes in salmon; insulin maintains its basal level but changes acutely in response to food intake, whereas IGF-I adjusts its basal levels to the long-term nutritional status and is less responsive to acute nutritional input. IGFBPs maintain their sensitivity to food intake, even after prolonged fasting, suggesting their critical role in the nutritional regulation of salmon growth.

  5. Genotypes and haplotypes in the insulin-like growth factors, their receptors and binding proteins in relation to plasma metabolic levels and mammographic density

    Directory of Open Access Journals (Sweden)

    Chanock Stephen J

    2010-03-01

    Full Text Available Abstract Background Increased mammographic density is one of the strongest independent risk factors for breast cancer. It is believed that one third of breast cancers are derived from breasts with more than 50% density. Mammographic density is affected by age, BMI, parity, and genetic predisposition. It is also greatly influenced by hormonal and growth factor changes in a woman's life cycle, spanning from puberty through adult to menopause. Genetic variations in genes coding for hormones and growth factors involved in development of the breast are therefore of great interest. The associations between genetic polymorphisms in genes from the IGF pathway on mammographic density and circulating levels of IGF1, its binding protein IGFBP3, and their ratio in postmenopausal women are reported here. Methods Samples from 964 postmenopausal Norwegian women aged 55-71 years were collected as a part of the Tromsø Mammography and Breast Cancer Study. All samples were genotyped for 25 SNPs in IGF1, IGF2, IGF1R, IGF2R, IGFALS and IGFBP3 using Taqman (ABI. The main statistical analyses were conducted with the PROC HAPLOTYPE procedure within SAS/GENETICS™ (SAS 9.1.3. Results The haplotype analysis revealed six haploblocks within the studied genes. Of those, four had significant associations with circulating levels of IGF1 or IGFBP3 and/or mammographic density. One haplotype variant in the IGF1 gene was found to be associated with mammographic density. Within the IGF2 gene one haplotype variant was associated with levels of both IGF1 and IGFBP3. Two haplotype variants in the IGF2R were associated with the level of IGF1. Both variants of the IGFBP3 haplotype were associated with the IGFBP3 level and indicate regulation in cis. Conclusion Polymorphisms within the IGF1 gene and related genes were associated with plasma levels of IGF1, IGFBP3 and mammographic density in this study of postmenopausal women.

  6. Determination of plasma protein binding rate of five components in Eucommia ulmoides extract%杜仲提取物中五个成分血浆蛋白结合率的测定

    Institute of Scientific and Technical Information of China (English)

    曹旭; 谢玉敏; 朱迪; 陈鹏程; 巩仔鹏; 王爱民

    2015-01-01

    目的:测定杜仲药材中5个成分的血浆蛋白结合率。方法采用平衡透析法测定血浆蛋白结合率,生物样本用甲醇沉淀蛋白进行处理,以葛根素为内标,利用超高效液相色谱-串联质谱仪测定血浆和缓冲溶液中的5个成分浓度。结果在所研究浓度范围内,京尼平苷酸、原儿茶酸、绿原酸、松脂醇二葡萄糖苷和松脂醇单葡萄糖苷的平均蛋白结合率分别为(25.77±2.68)%、(57.54±3.79)%、(53.91±3.00)%、(24.15±4.92)%、(49.78±3.61)%。结论京尼平苷酸和松脂醇二葡萄糖苷的蛋白结合率较低,原儿茶酸、绿原酸和松脂醇单葡萄糖苷则与大鼠血浆蛋白有中等强度的结合。%Aim To determine the plasma protein binding rate of five components of Eucommia ulmoides extract. Methods The equilibrium dialysis method was used to study the plasma protein binding rate. The plasma samples were extracted by protein precipitation with methanol. With the use of puerarin as the internal standard, UPLC-MS/MS was carried out to determine the concentration of the five compounds in and out of the dialysis membrane. Results The average plasma protein binding rates of five compounds on the area of the concentration which was determinate were as fol-lows, respectively: geniposidic acid was ( 25. 77 ± 2. 68 )%, protocatechuic acid was ( 57. 54 ± 3. 79)%, chlorogenic acid was (53. 91 ± 3. 00)%, pinoresinol diglucoside was (24. 15 ± 4. 92)%, and pinoresinol singleglucoside was (49. 78 ± 3. 61)%. Conclusions The results show that the binding percentage of geniposidic acid and pinoresinol diglucoside is relatively low, but the binding rate of the others with rat plasma protein is moderate.

  7. Determination of protein binding rates of celastrol in different species of plasma by HPLC%雷公藤红素在不同血浆中的蛋白结合率测定

    Institute of Scientific and Technical Information of China (English)

    袁菱; 陈彦; 周蕾; 郑丽燕; 曹伟

    2013-01-01

    OBJECTIVE To study the protein binding rate of celastrol in the plasma of rats, Beagle dogs and humans. METHODS Equilibrium dialysis method was carried out to determine the plasma protein binding rates in different species. HPLC was employed to determine the concentration of celastrol inside or outside of the dialysis membrane, and then protein binding rates were calculated. RESULTS The plasma protein binding rates of celastrol at low, middle and high concentrations in different species were as follows, the plasma protein binding rates were (78. 5 ± 1. 6) %, (77. 3 ± 2. 2) %, (76. 4 ± 2. 9) % in rats, (69.6±4.7)%, (71.1±3.4)%, (68. 0 ±2. 1)% in Beagle dogs, and (85. 6 ± 2. 7)%, (84.0±4. 7)%, (83.1±2.8)% in humans, respectively. CONCLUSION The protein binding rates of celastrol were of middle strength, which in rats, Beagle dogs and humans plasma were decreased in the following order: humans plasma>rats plasma>Beagle dogs plasma.%目的:研究雷公藤红素与大鼠、比格犬和人的血浆蛋白结合率.方法:采用血浆平衡透析法,以高效液相色谱法对透析内液与外液中雷公藤红素的含量进行测定,计算雷公藤红素与大鼠、比格犬和人的血浆蛋白结合率.结果:雷公藤红素低中高(0.25,0.5,1.0 μg·mL-1)3个浓度的大鼠血浆蛋白结合率分别为(78.5±1.6)%,(77.3±2.2)%,(76.4±2.9)%,比格犬血浆蛋白结合率分别为(69.6±4.7)%,(71.1±3.4)%,(68.0±2.1)%,人血浆蛋白结合率分别为(85.0±2.7)%,(84.0±4.7)%,(83.1±2.8)%.结论:雷公藤红素与血浆蛋白具有中等强度的结合,其中与人血浆中的蛋白结合较强,且人>大鼠>比格犬.

  8. Effect of seminal plasma zinc–binding proteins on motility and membrane integrity of canine spermatozoa stored at 5°C

    OpenAIRE

    Mogielnicka-Brzozowska Marzena; Dziekońska Anna; Strzeżek Rafał; Załęcki Michał; Majewska Anna; Tołścik Karolina; Kordan Władysław

    2014-01-01

    Sperm surface binding sites for non-zinc-binding proteins (nZnBPs) and zinc-binding proteins (ZnBPs) were studied by the fluorescence technique with biotin-labelled proteins. The nZnBPs binding pattern was unspecific, no characteristic sites on plasmalemma were found. ZnBPs were attached mainly to the acrosomal region of sperm head and to the sperm flagellum. ZnBPs added to the incubation mixture of the canine spermatozoa allowed the preservation of higher values for total motility, progressi...

  9. Plasma protein binding characteristics of triptolide in several species%雷公藤内酯醇与不同种属血浆蛋白结合率的研究

    Institute of Scientific and Technical Information of China (English)

    刘萍霞; 王娟; 庄笑梅; 张玉杰; 张振清; 阮金秀

    2012-01-01

    目的 考察雷公藤内酯醇在不同种属的血浆蛋白结合率.方法 超滤法分离结合型药物和游离型药物,LC-MS/MS法测定药物的浓度.结果 雷公藤内酯醇在不同浓度下与大鼠血浆的蛋白结合率分别是(25.4±1.8)、(12.7±8.5)和(11.5.±3.2)ng/ml,与比格犬血浆的蛋白结合率分别是(16.1±2.2)、(28.6±0.3)和(25.0±1.2)ng/ml;与人血浆的蛋白结合率分别是(21.5±2.1)、(18.9±5.7)和(21.3±2.6) ng/ml.结论 雷公藤内酯醇在实验浓度范围内与大鼠、比格犬和人血浆中蛋白结合率均较低,相同浓度雷公藤内酯醇与不同种属血浆蛋白结合率,以犬血浆最高[(28.6±0.3)ng/ml],与大鼠[(25.4±1.8) ng/ml]、人血浆[(18.9±5.7) g/ml]相比均具有统计学差异(P<0.05),大鼠与人血浆之间无统计学差异.各种属血浆蛋白结合率与药物浓度无明显依赖关系.%Objective To study the regularity of the plasma protein binding rate of triptolide ( TP ) in several species. Methods Ultrafiltration was employed to determine the plasma protein binding rate of triptolide in several species. Results The plasma protein binding rate of TP was 25.4 ± 1. 8, 12. 7 ±8. 5 and 11. 5 ±3.2 ng/ml in rat, 16. 1 ±2.2, 28.6 ±0.3 and 25. 0 ± 1.2 ng/ml in Beagles and 21. 5 ±2.1, 18. 9 ±5. 7 and 21. 3 ±2. 6 ng/ml in humans, respectively. Conclusion TP has a lower plasma protein binding rate in several species and the highest binding rate is showed in the dog plasma at the same concentration of 40 ng/ml. There is statistic difference between dog, rat and human plasma ( P <0. 05 ). The protein binding rates are not proportionally dependent on plasma concentrations of TP.

  10. Tetrabromobisphenol A Is an Efficient Stabilizer of the Transthyretin Tetramer.

    Directory of Open Access Journals (Sweden)

    Irina Iakovleva

    Full Text Available Amyloid formation of the human plasma protein transthyretin (TTR is associated with several human disorders, including familial amyloidotic polyneuropathy (FAP and senile systemic amyloidosis. Dissociation of TTR's native tetrameric assembly is the rate-limiting step in the conversion into amyloid, and this feature presents an avenue for intervention because binding of an appropriate ligand to the thyroxin hormone binding sites of TTR stabilizes the native tetrameric assembly and impairs conversion into amyloid. The desired features for an effective TTR stabilizer include high affinity for TTR, high selectivity in the presence of other proteins, no adverse side effects at the effective concentrations, and a long half-life in the body. In this study we show that the commonly used flame retardant tetrabromobisphenol A (TBBPA efficiently stabilizes the tetrameric structure of TTR. The X-ray crystal structure shows TBBPA binding in the thyroxine binding pocket with bromines occupying two of the three halogen binding sites. Interestingly, TBBPA binds TTR with an extremely high selectivity in human plasma, and the effect is equal to the recently approved drug tafamidis and better than diflunisal, both of which have shown therapeutic effects against FAP. TBBPA consequently present an interesting scaffold for drug design. Its absorption, metabolism, and potential side-effects are discussed.

  11. Inhibition of Human Transthyretin Aggregation by Non-Steroidal Anti-Inflammatory Compounds: A Structural and Thermodynamic Analysis

    Directory of Open Access Journals (Sweden)

    Luis Mauricio T. R. Lima

    2013-03-01

    Full Text Available Transthyretin (TTR is a homotetrameric protein that circulates in plasma and cerebral spinal fluid (CSF whose aggregation into amyloid fibrils has been associated with at least two different amyloid diseases: senile systemic amyloidosis (SSA and familial amyloid polyneuropathy (FAP. In SSA aggregates are composed of WT-TTR, while in FAP more than 100 already-described variants have been found in deposits. Until now, TTR-related diseases have been untreatable, although a new drug called Tafamidis has been approved only in Europe to specifically treat V30M patients. Thus, new strategies are still necessary to treat FAP caused by other variants of TTR. TTR has two channels in the dimer interface that bind to the hormone thyroxin and that have been used to accommodate anti-amyloidogenic compounds. These compounds stabilize the tetramers, rendering TTR less amyloidogenic. Here, we investigated the effects of three non-steroidal anti-inflammatory compounds—sulindac (SUL, indomethacin (IND and lumiracoxib (LUM—as tetramer stabilizers and aggregation inhibitors. WT-TTR and the very aggressive TTR variant L55P were used as models. These compounds were able to stabilize TTR against high hydrostatic pressure (HHP, increasing the ΔGf by several kcal. They were also effective in inhibiting WT-TTR and L55P acid- or HHP-induced aggregation; in particular, LUM and IND were very effective, inhibiting almost 100% of the aggregation of both proteins under certain conditions. The species formed when aggregation was performed in the presence of these compounds were much less toxic to cells in culture. The crystal structures of WT-TTR bound to the three compounds were solved at high resolution, allowing the identification of the relevant protein:drug interactions. We discuss here the ligand-binding features of LUM, IND and SUL to TTR, emphasizing the critical interactions that render the protein more stable and less amyloidogenic.

  12. Characterization of equine vitamin D–binding protein, development of an assay, and assessment of plasma concentrations of the protein in healthy horses and horses with gastrointestinal disease

    DEFF Research Database (Denmark)

    Pihl, Tina Holberg; Jacobsen, Stine; Olsen, Dorthe T.

    2017-01-01

    spectrometry. Monoclonal and polyclonal antibodies were raised against equine VDBP, and a rocket immunoelectrophoresis assay for equine VDBP was established. Plasma samples from 61 healthy horses were used to establish working VDBP reference values for study purposes. Plasma VDBP concentrations were assessed...

  13. Megalin binds and mediates cellular internalization of folate binding protein

    DEFF Research Database (Denmark)

    Birn, Henrik; Zhai, Xiaoyue; Holm, Jan

    2005-01-01

    Folate is an essential vitamin involved in a number of biological processes. High affinity folate binding proteins (FBPs) exist both as glycosylphosphatidylinositol-linked, membrane associated folate binding proteins and as soluble FBPs in plasma and some secretory fluids such as milk, saliva...... to bind and mediate cellular uptake of FBP. Surface plasmon resonance analysis shows binding of bovine and human milk FBP to immobilized megalin, but not to low density lipoprotein receptor related protein. Binding of (125)I-labeled folate binding protein (FBP) to sections of kidney proximal tubule, known...

  14. Determination of the protein binding rates of apigenin with different kinds of plasma by HPLC%高效液相色谱法测定芹菜素在不同血浆中的蛋白结合率

    Institute of Scientific and Technical Information of China (English)

    任非; 段坤峰; 吴宗耀; 王蓓; 杨静; 陈学军

    2012-01-01

    目的:建立测定芹菜素在大鼠血浆、人血浆和牛血清白蛋白(BSA)中蛋白结合率的方法,并计算不同介质中的的相关参数.方法:采用高效液相色谱(HPLC)测定不同介质中芹菜素的浓度,并结合平衡透析法测定蛋白结合率.结果:高、中、低浓度下,芹菜素的血浆蛋白结合率分别为:大鼠血浆:(99.4±3.8)%、(99.6±5.6)%、(99.5±4.5)%;人血浆:(96.3±7.2)%、(97.9±10.5)%、(97.8±9.7)%;牛血清白蛋白:(98.7±8.6)%、(99.6±9.4)%、(99.1±8.0)%.结论:本方法快速、简便、可靠,芹菜素与大鼠血浆、人血浆和牛血清白蛋白结合率很高,且与血药浓度无显著相关性.%OBJECTIVE To establish a method for determining the protein binding rates of apigenin in rat plasma, human plasma.bovine serum albumin (BSA).and to calculate the correlate related parameters of apigenin in different matrix. METHODS The concentrations of apigenin in different matrix were assayed by high performance liquid chromatography (HPl.C). The protein binding rates of apigenin in different matrix were determined by equilibrium dialysis method. RESULTS The protein binding rates of apigenin with rat plasma,human plasma and BSA were (99. 4 ± 3.8) %. (99. 6 ± 5. 6) %, (99. 5 ± 4. 5) %, respectively, when high concentration of apigenin was used;they were (96. 3 ± 7. 2)%, (97. 9 + 10. 5)%. (97. 8 ± 9. 7)% .respectively, when medium concentration of apigenin was used;and they were (98. 7 ± 8. 6)%, (99. 6 + 9. 4)%, (99. 1 +8.0)%, respectively,when low concentration of apigenin was used CONCLUSION The method was validated to be rapid.and reliable. The protein binding rates of apigenin with rat plasma, human plasma and BSA were very high. Moreover, the plasma protein binding rates were dose-independent.

  15. 冬凌草甲素在不同种属血浆中蛋白结合率的HPLC法测定%HPLC determination of protein binding rates of oridonin different plasma

    Institute of Scientific and Technical Information of China (English)

    杜英峰; 靳怡然; 许慧君; 刘彭玮; 张兰桐; 陈玉侠; 王丽亮

    2012-01-01

    目的:建立冬凌草甲素在大鼠血浆、人血浆和牛血清白蛋白中蛋白结合率的测定方法,并计算不同种属血浆蛋白的相关参数.方法:采用HPLC法测定血浆中药物总浓度及游离药物浓度,应用平衡透析法测定蛋白结合率.结果:大鼠血浆中冬凌草甲素高、中、低3个浓度的血浆蛋白结合率分别为(69.66±12.8)%,(59.62±12.6)%,(57.94±4.1)%;人血浆中冬凌草甲素高、中、低3个浓度的血浆蛋白结合率分别为(78.15±3.6)%,(77.92±8.8)%,(76.72±7.3)%;牛血清白蛋白中冬凌草甲素高、中、低3个浓度的血浆蛋白结合率分别为(35.58±7.2)%,(34.59±10.8)%,(32.03±6.0)%.结论:在体外冬凌草甲素与大鼠血浆、人血浆和牛血清白蛋白属中等结合型药物,且蛋白结合率随着药物血浆浓度的增加无明显的浓度依赖性.%Objective: To develop an HPLC method to determine the protein binding rates of oridonin in human plasma, rat plasma and bovine serum albumin (BSA) , and to calculate related parameters of oridonin to different genera plasma proteins. Methods: The binding rates of oridonin with different genera plasma proteins were determined by equilibrium dialysis method. The concentrations of oridonin were assayed by HPLC. Results;The binding rates of oridonin with rat, human plasma and BSA at three concentrations levels were (69.66 ± 12. 8) % , (59.62 ± 12.6)%, (57.94 ±4. 1)%;(78. 15 ±3.6)% ,(77.92 ±8. 8)% ,(76.72 ±7.3)% and(35. 58 ±7. 2)% ,(34.59 ± 10. 8)% , (32.03 ±6.0)% ,respectively. Conclusion;The binding rates of oridonin with human,rat plasma and BSA are middle. Moreover,the binding rates are not dependent proportionally to plasma concentration of oridonin.

  16. Nuclear Receptor HNF4α Binding Sequences are Widespread in Alu Repeats

    Directory of Open Access Journals (Sweden)

    Bolotin Eugene

    2011-11-01

    Full Text Available Abstract Background Alu repeats, which account for ~10% of the human genome, were originally considered to be junk DNA. Recent studies, however, suggest that they may contain transcription factor binding sites and hence possibly play a role in regulating gene expression. Results Here, we show that binding sites for a highly conserved member of the nuclear receptor superfamily of ligand-dependent transcription factors, hepatocyte nuclear factor 4alpha (HNF4α, NR2A1, are highly prevalent in Alu repeats. We employ high throughput protein binding microarrays (PBMs to show that HNF4α binds > 66 unique sequences in Alu repeats that are present in ~1.2 million locations in the human genome. We use chromatin immunoprecipitation (ChIP to demonstrate that HNF4α binds Alu elements in the promoters of target genes (ABCC3, APOA4, APOM, ATPIF1, CANX, FEMT1A, GSTM4, IL32, IP6K2, PRLR, PRODH2, SOCS2, TTR and luciferase assays to show that at least some of those Alu elements can modulate HNF4α-mediated transactivation in vivo (APOM, PRODH2, TTR, APOA4. HNF4α-Alu elements are enriched in promoters of genes involved in RNA processing and a sizeable fraction are in regions of accessible chromatin. Comparative genomics analysis suggests that there may have been a gain in HNF4α binding sites in Alu elements during evolution and that non Alu repeats, such as Tiggers, also contain HNF4α sites. Conclusions Our findings suggest that HNF4α, in addition to regulating gene expression via high affinity binding sites, may also modulate transcription via low affinity sites in Alu repeats.

  17. Influence of one- and two-dimensional gel electrophoresis procedure on metal-protein bindings examined by electrospray ionization mass spectrometry, inductively coupled plasma mass spectrometry, and ultrafiltration.

    Science.gov (United States)

    Schmidt, Anne-Christine; Störr, Bianca; Kummer, Nicolai-Alexeji

    2011-08-15

    Three independent methods, (i) electrospray ionization mass spectrometry (ESI-MS), (ii) carrying out the complete protein preparation procedure required for protein gel electrophoresis (GE) including extraction, precipitation, washing, and desalting with subsequent microwave digestion of the produced protein fractions for metal content quantification, and (iii) ultrafiltration for separating protein-bound and unbound metal fractions, were employed to elucidate the influences of protein sample preparation and GE running conditions on metal-protein bindings. A treatment of the protein solution with acetone instead of trichloroacetic acid or ammonium sulfate for precipitate formation led to a strongly enhanced metal binding capacity. The desalting step of the resolubilized protein sample caused a metal loss between 10 and 35%. The omission of some extraction buffer additives led to a diminished metal binding capacity of protein fractions obtained from the sample preparation procedure for GE, whereas a tenside addition to the protein solution inhibited metal-protein bindings. The binding stoichiometry of Cu and Zn-protein complexes determined by ESI-MS was influenced by the type of the metal salt which was applied to the protein solution. A higher pH value of the sample solution promoted the metal ion complexation by the proteins. Ultrafiltration experiments revealed a higher Cu- and Zn-binding capacity of the model protein lysozyme in both resolubilization buffers for 1D- and 2D-GE compared to the protein extraction buffer. Strongly diminished metal binding capacities of lysozyme were recorded in the running buffer of 1D-GE and in the gel staining solutions.

  18. 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.

  19. Increase in Skin Autofluorescence and Release of Heart-Type Fatty Acid Binding Protein in Plasma Predicts Mortality of Hemodialysis Patients

    NARCIS (Netherlands)

    Arsov, Stefan; Trajceska, Lada; van Oeveren, Wim; Smit, Andries J.; Dzekova, Pavlina; Stegmayr, Bernd; Sikole, Aleksandar; Rakhorst, Gerhard; Graaff, Reindert

    2013-01-01

    Advanced glycation end-products (AGEs) are uremic toxins that accumulate progressively in hemodialysis (HD) patients. The aim of this study was to assess the 1-year increase in skin autofluorescence (DAF), a measure of AGEs accumulation and plasma markers, as predictors of mortality in HD patients.

  20. Collectin 11 (CL-11, CL-K1) is a MASP-1/3-associated plasma collectin with microbial-binding activity

    DEFF Research Database (Denmark)

    Hansen, Soren; Selman, Lana; Palaniyar, Nades

    2010-01-01

    and showed that CL-11 has preference for l-fucose and d-mannose. We finally show that CL-11 binds to intact bacteria, fungi, and viruses and that CL-11 decreases influenza A virus infectivity and forms complexes with DNA. On the basis of the significant concentration of CL-11 in circulation and CL-11's...

  1. Determination of protein binding rate of salicylic acid in plasma derived from different species via high-per-formance liquid chromatography%水杨酸在不同血浆中的蛋白结合率测定

    Institute of Scientific and Technical Information of China (English)

    张农山; 张立; 王培民

    2014-01-01

    Objective To investigate the protein binding ratio of salicylic acid in the plasma derived from rats, Beagle dogs and human. Methods Equilibrium dialysis was performed to determine the plasma protein binding rates in plasma derived from different species. High-performance liquid chromatography (HPLC) was employed to determine the concentration of salicylic acid inside or outside of the dialysis membrane, which allowed the calculation of protein binding rates. Results The plasma protein binding rates of salicylic acid at low, middle and high concentrations in different species were (91.1±1.4)%, (90.2±2.5)% and (90.7±3.1)% in rats, (82.5±2.1)%, (81.3±1.8)% and (83.1±2.3)%in Beagle dogs, and (96.5±2.7)%, (97.2±3.0)% and (96.4±1.6)% in humans, respectively. Conclusion The protein binding rates of salicylic acid are highest in human, followed by rats and Beagle dogs.%目的:研究水杨酸与大鼠、比格犬和人的血浆蛋白结合率。方法采用平衡透析法,以高效液相色谱法对透析内液与外液中水杨酸的含量进行测定,计算水杨酸与大鼠、比格犬和人的血浆蛋白结合率。结果水杨酸低、中、高(0.6,1,3μg/ml)3个浓度的大鼠血浆蛋白结合率分别为(91.1±1.4)%,(90.2±2.5)%,(90.7±3.1)%;比格犬血浆蛋白结合率分别为(82.5±2.1)%,(81.3±1.8)%,(83.1±2.3)%;人血浆蛋白结合率分别为(96.5±2.7)%,(97.2±3.0)%,(96.4±1.6)%。结论水杨酸与血浆蛋白具有高强度的结合,其中与人血浆中的蛋白结合较强,且人>大鼠>比格犬。

  2. Relationship between seminal plasma zinc concentration and spermatozoa–zona pellucida binding and the ZP-induced acrosome reaction in subfertile men

    OpenAIRE

    Liu, De-Yi; Sie, Boon-Shih; Liu, Ming-Li; Agresta,Franca; Baker, HW Gordon

    2009-01-01

    The aim of this study was to determine the relationship between seminal zinc concentration and spermatozoa–zona pellucida (ZP) binding and the ZP-induced acrosome reaction (ZPIAR) in subfertile men. Semen analyses and seminal zinc concentration assessments were carried out according to the World Health Organization manual for 458 subfertile men. A spermatozoa–ZP interaction test was carried out by incubating 2 × 106 motile spermatozoa with a group of four unfertilized oocytes obtained from a ...

  3. Proteomic analysis of heparin-binding proteins from human seminal plasma: a step towards identification of molecular markers of male fertility

    Indian Academy of Sciences (India)

    Vijay Kumar; Md Imtaiyaz Hassan; Anil Kumar Tomar; Tara Kashav; Jaya Nautiyal; Sarman Singh; Tej P Singh; Savita Yadav

    2009-12-01

    Glycosaminoglycans, especially heparin, are involved in various cell processes such as apoptosis, cell cycle control, platelet activation, capacitation, acrosome reaction and sperm decondensation. Heparin-binding proteins (HBPs) are essential constituents of human seminal fluid, which bind to sperm lipids containing the phosphorylcholine group and mediate the fertilization process. We utilized a proteomic set-up consisting of affinity chromatography, isoelectric focusing (IEF) coupled with matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (MALDI TOF/MS) for protein analysis of human HBPs. We resolved 70 different spots on two-dimensional (2-D) gel and subsequently identified these proteins. Forty different types of proteins were identified. Functional analysis revealed that 38% of the proteins belonged to the enzyme category, 20% were involved in RNA processing and transcription, 18% in structure and transport function, and 16% in cell recognition and signal transduction. We also identified 8% of proteins with unknown functions, although their expression in seminal fluid has been documented. Proteins of seminal fluid that bind heparin may be directly involved in sperm capacitation and acrosome reaction (AR), which are the two critical steps for fertilization. This information on HBPs would be useful for identifying potential biomarkers of fertility in the near future.

  4. Sardinian Type 1 diabetes patients, Transthyretin and Mycobacterium avium subspecies paratuberculosis infection

    Directory of Open Access Journals (Sweden)

    Masala Speranza

    2012-12-01

    Full Text Available Abstract Background Mycobacterium avium subsp. paratuberculosis (MAP is the cause of Johne’s disease, an enteric granulomatous disease. Recently, MAP has been associated with different autoimmune diseases such as Crohn’s disease, type 1 diabetes (T1D and multiple sclerosis. Transthyretin (TTR is a plasma transport protein for thyroid hormone and forms a complex with retinol-binding protein. Reduced TTR plasma levels in MAP infected ovines have been reported. TTR exerts also a functional role in the pancreas promoting insulin release and protecting β-cells from death. Our objective was to identify a protein that could be used as a diagnostic marker of T1D for determining disease progression and monitoring at-risk patients. We postulate that serological TTR levels would be reduced in T1D MAP exposed patients. Our hypothesis is based on the observation of cases of T1D patients with decreased TTR levels beside the reduced TTR plasma levels in ovines with Johne’s disease. We quantified the plasma protein levels of TTR in 50 people with T1D and 51 age-matched healthy controls (HCs by means of enzyme-linked immunosorbent assays (ELISA. Findings Our pilot study showed that plasma TTR levels were not significantly lower/higher in T1D Sardinian cases compared to the HCs. Conclusion These preliminary data indicate that plasma TTR may not be a good candidate biomarker for T1D diagnosis and further studies to elucidate the possible link are needed.

  5. Relationship between seminal plasma zinc concentration and spermatozoa-zona pellucida binding and the ZP-induced acrosome reaction in subfertile men.

    Science.gov (United States)

    Liu, De-Yi; Sie, Boon-Shih; Liu, Ming-Li; Agresta, Franca; Baker, H W Gordon

    2009-07-01

    The aim of this study was to determine the relationship between seminal zinc concentration and spermatozoa-zona pellucida (ZP) binding and the ZP-induced acrosome reaction (ZPIAR) in subfertile men. Semen analyses and seminal zinc concentration assessments were carried out according to the World Health Organization manual for 458 subfertile men. A spermatozoa-ZP interaction test was carried out by incubating 2 x 10(6) motile spermatozoa with a group of four unfertilized oocytes obtained from a clinical in vitro fertilization programme. After 2 h of incubation, the number of spermatozoa bound per ZP and the ZPIAR of ZP-bound spermatozoa were examined. The effect of adding 0.5 mmol L(-1) zinc to the media on the ZPIAR of spermatozoa from normozoospermic men was also tested in vitro. Seminal zinc concentration positively correlated with sperm count and duration of abstinence, but negatively correlated with semen volume. On analysis of data from all participants, both spermatozoa-ZP binding and the ZPIAR were significantly correlated with sperm motility and normal morphology, but not with seminal zinc concentration. However, in men with normozoospermic semen, the seminal zinc concentration was significantly higher in men with defective ZPIAR (or= 16%) (P zinc to the culture media had no effect on spermatozoa-ZP binding, but significantly reduced the ZPIAR in vitro (P zinc concentration is correlated with sperm count and the duration of abstinence in subfertile men. In men with normozoospermic semen, high seminal zinc concentration may have an adverse effect on the ZPIAR.

  6. The aging changes of the plasma retinol-binding protein 4 levels in elderly essential hypertension patients%老年高血压患者血浆视黄醇结合蛋白的增龄变化

    Institute of Scientific and Technical Information of China (English)

    曹萍; 李睿; 沈丹; 钟亚

    2011-01-01

    Objective To explore the aging changes of the plasma retinol-binding protein 4(RBP4)levels in elderly essential hypertension patients and to evaluate the influential factors. Methods 275 essential hypertension patients were divided into two groups according to whether complicated by type 2 diabetes mellitus. Fifty healthy persons served as normal control group. Plasma RBP4 levels were determined in the three groups. The relationships of plasma RBP4 levels with the blood pressure,the plasma lipids levels and body mass index were analyzed. The effect of aging in the 275 patients on the plasma RBP4 levels was explored. Results The plasma RBP4 levels were significantly increased both in the pure hypertension group and in the hypertension complicated by type 2 diabetes mellitus group compared with the control group (P<0.01), but there was no statistical difference between the pure hypertension group and the hypertension complicated by type 2 diabetes mellitus group. The level of plasma RBP4 was positively correlated with plasma triglyceride and low density lipoprotein-cholesterol (r = 0. 347, r = 0. 229, P < 0.01). However, the level of plasma RBP4 gradually decreased with age in all essential hypertension patients. The age was negatively correlated with the level of plasma RBP4, buttock circumference, body weight, body mass index, diastolic pressure,average arterial pressure, triglyceride and low density lipoprotein cholesterol (P < 0.01). Conclusion There is a phenomenon of insulin resistance which correlates with plasma RBP4 increase in the patients with essential hypertension. The level of plasma RBP4 gradually decreases with age in all essential hypertension patients. It is inferred that the insulin resistance plays a more important part in essential hypertension in young patients than in elderly patients.%目的 探讨老年高血压患者血浆视黄醇结合蛋白(retionl binding protein 4,RBP4)水平的增龄变化及其影响因素.方法 将275例高

  7. Platinum group metallodrug-protein binding studies by capillary electrophoresis - inductively coupled plasma-mass spectrometry: a further insight into the reactivity of a novel antitumor ruthenium(III) complex toward human serum proteins.

    Science.gov (United States)

    Polec-Pawlak, Kasia; Abramski, Jan K; Semenova, Olga; Hartinger, Christian G; Timerbaev, Andrei R; Keppler, Bernhard K; Jarosz, Maciej

    2006-03-01

    Biochemical speciation analysis has become a hot area of CE research due largely to growing emergence of inductively coupled plasma (ICP)-MS as a proper detection technique. A benefit of CE-ICP-MS coupling in species-selective analysis of anticancer metal-based drugs is the possibility of distinguishing the signals of the intact drug and its metabolites and hence of quantifying them independently. This advantage (over CE with UV-vis detection) was exploited here in order to gain better knowledge about the rate and degree of the transformation of indazolium [trans-tetrachlorobis(1H-indazole)ruthenate(III)] (KP1019), a promising tumor-inhibiting agent that successfully finished phase I clinical studies, upon its binding toward individual serum transport proteins. At increasing the KP1019/protein molar ratio, the reaction rate expressed by an evolving peak of the protein adduct became faster, with the equilibrium state being reached after about 40 and 60 min of incubation at 37 degrees C for transferrin and albumin, respectively. The binding reaction was shown to obey the first-order character that enabled for reliable calculation of the corresponding rate constants as (28.7 +/- 1.5) x 10(-4) and (10.6 +/- 0.7) x 10(-4)/s, respectively. When incubated with a ten-fold excess of KP1019, albumin and transferrin bound, respectively, up to 8 and 10 equiv. of ruthenium (Ru). Relative affinity of KP1019 toward different proteins under simulated physiological conditions was also characterized in terms of the overall binding constants (5600 and 10 600/M, respectively). To emphasize the difference in the protein-binding behavior, a competitive interaction of KP1019 was followed by CE-ICP-MS at the actual molar ratio of proteins in blood, i.e. a ten-fold excess of albumin over transferrin. The fact that KP1019 binds to albumin stronger than to transferrin was manifested by finding almost all ruthenium (98-99%) in the albumin fraction.

  8. [The Role of Membrane-Bound Heat Shock Proteins Hsp90 in Migration of Tumor Cells in vitro and Involvement of Cell Surface Heparan Sulfate Proteoglycans in Protein Binding to Plasma Membrane].

    Science.gov (United States)

    Snigireva, A V; Vrublevskaya, V V; Skarga, Y Y; Morenkov, O S

    2016-01-01

    Heat shock protein Hsp90, detected in the extracellular space and on the membrane of cells, plays an important role in cell motility, migration, invasion and metastasis of tumor cells. At present, the functional role and molecular mechanisms of Hsp90 binding to plasma membrane are not elucidated. Using isoform-specific antibodies against Hsp90, Hsp9α and Hsp90β, we showed that membrane-bound Hsp90α and Hsp90β play a significant role in migration of human fibrosarcoma (HT1080) and glioblastoma (A-172) cells in vitro. Disorders of sulfonation of cell heparan sulfates, cleavage of cell heparan. sulfates by heparinase I/III as well as treatment of cells with heparin lead to an abrupt reduction in the expression level of Hsp90 isoforms. Furthermore, heparin significantly inhibits tumor cell migration. The results obtained demonstrate that two isoforms of membrane-bound Hsp90 are involved in migration of tumor cells in vitro and that cell surface heparan sulfate proteoglycans play a pivotal role in the "anchoring" of Hsp90α and Hsp90β to the plasma membrane.

  9. Analysis of metal-binding proteins separated by non-denaturating gel electrophoresis using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS).

    Science.gov (United States)

    Becker, J Susanne; Mounicou, Sandra; Zoriy, Miroslav V; Becker, J Sabine; Lobinski, Ryszard

    2008-09-15

    Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) have become established as very efficient and sensitive biopolymer and elemental mass spectrometric techniques for studying metal-binding proteins (metalloproteins) in life sciences. Protein complexes present in rat tissues (liver and kidney) were separated in their native state in the first dimension by blue native gel electrophoresis (BN-PAGE). Essential and toxic metals, such as zinc, copper, iron, nickel, chromium, cadmium and lead, were detected by scanning the gel bands using quadrupole LA-ICP-MS with and without collision cell as a microanalytical technique. Several proteins were identified by using MALDI-TOF-MS together with a database search. For example, on one protein band cut from the BN-PAGE gel and digested with the enzyme trypsin, two different proteins - protein FAM44B and cathepsin B precursor - were identified. By combining biomolecular and elemental mass spectrometry, it was possible to characterize and identify selected metal-binding rat liver and kidney tissue proteins.

  10. [Transthyretin: it's miracle function and pathogenesis].

    Science.gov (United States)

    Ando, Yukio

    2009-03-01

    Transthyretin (TTR) was previously called prealbumin because the band it formed on agarose gel electrophoresis at pH 8.6 was at the prealbumin position. However, it has been well documented that TTR of rodents does not show a prealbumin position on electrophoresis. Now, its name describes its function, binding to retinol binding protein (RBP) and T4. The serum concentration of the protein is 20-40 mg/dl, and TTR forms a tetramer. The plasma half life of the protein is 1.9 days. TTR is synthesized by the liver, retina, pancreas, and choroid plexus. In cerebro-spinal fluid (CSF), it is the second most abundant protein, and is considered as an important protein in the pathogenesis of Alzheimer's disease, depression, and lead intoxication. In addition, TTR is a tryptophan-rich protein, it is used as one of the nutrition assessment proteins, it acts as an anti acute phase protein, and its plasma concentration decreases during inflammation and bacterial infection. Since TTR is a highly amyloidogenic protein because it contains a beta-sheet structure, it becomes a precursor protein in familial amyloidotic polyneuropathy(FAP). Moreover, TTR plays important roles in various CNS disorders, diabetes melitus, and lipid metabolism.

  11. MD-2 binds cholesterol.

    Science.gov (United States)

    Choi, Soo-Ho; Kim, Jungsu; Gonen, Ayelet; Viriyakosol, Suganya; Miller, Yury I

    2016-02-19

    Cholesterol is a structural component of cellular membranes, which is transported from liver to peripheral cells in the form of cholesterol esters (CE), residing in the hydrophobic core of low-density lipoprotein. Oxidized CE (OxCE) is often found in plasma and in atherosclerotic lesions of subjects with cardiovascular disease. Our earlier studies have demonstrated that OxCE activates inflammatory responses in macrophages via toll-like receptor-4 (TLR4). Here we demonstrate that cholesterol binds to myeloid differentiation-2 (MD-2), a TLR4 ancillary molecule, which is a binding receptor for bacterial lipopolysaccharide (LPS) and is indispensable for LPS-induced TLR4 dimerization and signaling. Cholesterol binding to MD-2 was competed by LPS and by OxCE-modified BSA. Furthermore, soluble MD-2 in human plasma and MD-2 in mouse atherosclerotic lesions carried cholesterol, the finding supporting the biological significance of MD-2 cholesterol binding. These results help understand the molecular basis of TLR4 activation by OxCE and mechanisms of chronic inflammation in atherosclerosis.

  12. Human plasma kallikrein and tissue kallikrein binding to a substrate based on the reactive site of a factor Xa inhibitor isolated from Bauhinia ungulata seeds.

    Science.gov (United States)

    Oliva, M L; Andrade, S A; Batista, I F; Sampaio, M U; Juliano, M; Fritz, H; Auerswald, E A; Sampaio, C A

    1999-12-01

    Kunitz type Bauhinia ungulata factor Xa inhibitor (BuXI) was purified from B. ungulata seeds. BuXI inactivates factor Xa and human plasma kallikrein (HuPK) with Ki values of 18.4 and 6.9 nM, respectively. However, Bauhinia variegata trypsin inhibitor (BvTI) which is 70% homologous to BuXI does not inhibit factor Xa and is less efficient on HuPK (Ki = 80 nM). The comparison between BuXI and BvTI reactive site structure indicates differences at Met59, Thr66 and Met67 residues. The hydrolysis rate of quenched fluorescence peptide substrates based on BuXI reactive site sequence, Abz-VMIAALPRTMFIQ-EDDnp (leading peptide), by HuPK and porcine pancreatic kallikrein (PoPK) is low, but hydrolysis is enhanced with Abz-VMIAALPRTMQ-EDDnp, derived from the leading peptide shortened by removing the dipeptide Phe-Ileu from the C-terminal portion, for HuPK (Km = 0.68 microM, k(cat)/Km = 1.3 x 10(6) M(-1) s(-1)), and the shorter substrate Abz-LPRTMQ-EDDnp is better for PoPK (Km = 0.66 microM, k(cat)/Km = 2.2 x 10(3) M(-1) s(-1)). The contribution of substrate methionine residues to HuPK and PoPK hydrolysis differs from that observed with factor Xa. The determined Km and k(cat) values suggest that the substrates interact with kallikreins the same as an enzyme and inhibitor interacts to form complexes.

  13. MASP2在小儿上呼吸道感染中的意义%Plasma levels of mannan-binding lectin-associated serine protease 2 in children with upper respiratory tract infection

    Institute of Scientific and Technical Information of China (English)

    熊思敏; 赵娜; 裘宇容; 张丽芸; 左大明; 陈政良

    2015-01-01

    Absteact:Objective To explore the significance of plasma levels of mannan-binding lectin (MBL)-associated serine protease 2 (MASP2) in children with upper respiratory tract infection (URTI). Methods A total of 103 children with URTI and 35 healthy children were examined for plasma levels of MASP2 and C-reactive protein (CRP). According to CRP levels, white blood cell count (WBC), stage of infection, and administration of treatments, the children with URTI were divided into the elevated CRP group (n=48) and the normal CRP group (n=54), elevated WBC group (n=61) and normal WBC group (n=40), the early stage of infection without treatment group (n=68) and mid-late stage of infection with treatment group (n=35). Results Plasma MASP2 levels was significantly higher in URTI group than in the healthy control group (P0.05), correlated with WBC in elevated WBC group (r=0.392, P0.05), and was significantly higher in early stage infection without treatment group than in mid-late stage of infection with treatment group (P<0.01). MASP2, MBL2 and CRP genes had a common binding site for the transcription factor HNF-4α. Conclusion MASP2 may be an acute-phase protein, and its plasma level might serve as a new reference index in the diagnosis of URTI in children.%目的:探讨血浆甘露聚糖结合凝集素(MBL)相关丝氨酸蛋白酶2(MASP2)水平在儿童上呼吸道感染(上感)中的意义。方法取103例上感患儿和35例健康体检儿童作为研究对象,测定其血浆MASP2和C反应蛋白(CRP)浓度并进行白细胞计数(WBC)。根据CRP和WBC值、感染不同时期及有无治疗,上感儿童分为CRP升高组(n=48)与正常组(n=54)、WBC升高组(n=61)与正常组(n=40)、感染早期未用药组(n=68)与感染中后期已用药组(n=35),对各组资料进行统计学分析。结果上感组MASP2浓度显著高于对照组(P<0.001),CRP升高组血浆MASP2浓度与CRP值正相关(r=0.310,P<0.05

  14. Localization of the ATP-binding cassette (ABC transport proteins PfMRP1, PfMRP2, and PfMDR5 at the Plasmodium falciparum plasma membrane

    Directory of Open Access Journals (Sweden)

    Luty Adrian JF

    2009-08-01

    Full Text Available Abstract Background The spread of drug resistance has been a major obstacle to the control of malaria. The mechanisms underlying drug resistance in malaria seem to be complex and multigenic. The current literature on multiple drug resistance against anti-malarials has documented PfMDR1, an ATP-binding cassette (ABC protein, as an important determinant of resistance. In the Plasmodium falciparum genome, there are several ABC transporters some of which could be putative drug transporting proteins. In order to understand the molecular mechanisms underlying drug resistance, characterization of these transporters is essential. The aim of this study was to characterize and localize putative ABC transporters. Methods In the plasmoDB database, 16 members of the P. falciparum ABC family can be identified, 11 of which are putative transport proteins. A phylogenetic analysis of the aligned NBDs of the PfABC genes was performed. Antibodies against PfMRP1 (PfABCC1, PfMRP2 (PfABCC2, and PfMDR5 (PfABCB5 were generated, affinity purified and used in immunocytochemistry to localize the proteins in the asexual stages of the parasite. Results The ABC family members of P. falciparum were categorized into subfamilies. The ABC B subfamily was the largest and contained seven members. Other family members that could be involved in drug transport are PfABCC1, PfABCC2, PfABCG1, and PfABCI3. The expression and localization of three ABC transport proteins was determined. PfMRP1, PfMRP2, and PfMDR5 are localized to the plasma membrane in all asexual stages of the parasite. Conclusion In conclusion, 11 of the 16 ABC proteins in the P. falciparum genome are putative transport proteins, some of which might be involved in drug resistance. Moreover, it was demonstrated that three of these proteins are expressed on the parasite's plasma membrane.

  15. Technetium-99m labeling and fibronectin binding ability of Corynebacterium diphtheriae; Marcacao de Corynebacterium diphtheriae com Tecnecio-99m e avaliacao da capacidade de ligacao a fibronectina de plasma humano

    Energy Technology Data Exchange (ETDEWEB)

    Souza, S.M.S.; Nagao, P.E.; Bernardo-Filho, M. [Universidade do Estado do Rio de Janeiro, RJ (Brazil). Inst. de Biologia Roberto Alcantara Gomes; Pereira, G.A.; Napoleao, F.; Andrade, A.F.B.; Hirata Junior, R.; Mattos-Guaraldi, A.L. [Universidade do Estado do Rio de Janeiro, RJ (Brazil). Faculdade de Ciencias Medicas

    2004-04-15

    The use of radionuclides has permitted advances in areas of clinical and scientific knowledge. Several molecules and cells have been labelled with Technetium-99m ({sup 99m}Tc). The stannous chloride (SnCl{sub 2}) has a significant influence on the labeling and stability of {sup 99m}Tc radiotracers. The frequent risk of diphtheria epidemics has intensified interest in the virulence factors of Corynebacterium diphtheriae. Although studies have looked at potential adhesins including haemagglutinins and exposed sugar residues, the molecular basis of mechanisms of adherence remains unclear. Adherence of pathogens to mammalian tissues may be mediated by fibronectin (FN) found in body fluids, matrix of connective tissues, and cell surfaces. In the present study we evaluated the binding ability to human plasma FN by {sup 99m}Tc labeled-C.diphtheriae. Due to adverse effects of stannous ions, microorganisms were submitted to survival and filamentation induction assays. Data showed a dose dependent susceptibility to SnCl{sub 2} bactericidal effects. Cell filamentation was observed for concentrations of SnCl{sub 2} > 110 {mu}g/ml. Adherence levels of {sup 99m}Tc labelled 241strain to coverslips coated with 20 {mu}g/ml FN were higher (P = 0.0037) than coated with bovine serum albumin. FN binding by the sucrose fermenting 241 C. diphtheriae strain (8.9% + 2.6) was significantly lower (P=0.0139) than Staphylococcus aureus Cowan I strain (34.1% {+-} 1.2). Therefore, bacterial {sup 99m}Tc labeling represents an additional tool that may contribute to the comprehension of C. diphtheriae interactions with host receptors such as FN that act as biological organizers by holding bacterial cells in position and guiding their migration. (author)

  16. Tight-binding quantum chemical molecular dynamics simulations for the elucidation of chemical reaction dynamics in SiC etching with SF6/O2 plasma.

    Science.gov (United States)

    Ito, Hiroshi; Kuwahara, Takuya; Kawaguchi, Kentaro; Higuchi, Yuji; Ozawa, Nobuki; Kubo, Momoji

    2016-03-21

    We used our etching simulator [H. Ito et al., J. Phys. Chem. C, 2014, 118, 21580-21588] based on tight-binding quantum chemical molecular dynamics (TB-QCMD) to elucidate SiC etching mechanisms. First, the SiC surface is irradiated with SF5 radicals, which are the dominant etchant species in experiments, with the irradiation energy of 300 eV. After SF5 radicals bombard the SiC surface, Si-C bonds dissociate, generating Si-F, C-F, Si-S, and C-S bonds. Then, etching products, such as SiS, CS, SiFx, and CFx (x = 1-4) molecules, are generated and evaporated. In particular, SiFx is the main generated species, and Si atoms are more likely to vaporize than C atoms. The remaining C atoms on SiC generate C-C bonds that may decrease the etching rate. Interestingly, far fewer Si-Si bonds than C-C bonds are generated. We also simulated SiC etching with SF3 radicals. Although the chemical reaction dynamics are similar to etching with SF5 radicals, the etching rate is lower. Next, to clarify the effect of O atom addition on the etching mechanism, we also simulated SiC etching with SF5 and O radicals/atoms. After bombardment with SF5 radicals, Si-C bonds dissociate in a similar way to the etching without O atoms. In addition, O atoms generate many C-O bonds and COy (y = 1-2) molecules, inhibiting the generation of C-C bonds. This indicates that O atom addition improves the removal of C atoms from SiC. However, for a high O concentration, many C-C and Si-Si bonds are generated. When the O atoms dissociate the Si-C bonds and generate dangling bonds, the O atoms terminate only one or two dangling bonds. Moreover, at high O concentrations there are fewer S and F atoms to terminate the dangling bonds than at low O concentration. Therefore, few dangling bonds of dissociated Si and C atoms are terminated, and they form many Si-Si and C-C bonds. Furthermore, we propose that the optimal O concentration is 50-60% because both Si and C atoms generate many etching products producing fewer C

  17. Binding Procurement

    Science.gov (United States)

    Rao, Gopalakrishna M.; Vaidyanathan, Hari

    2007-01-01

    This viewgraph presentation reviews the use of the binding procurement process in purchasing Aerospace Flight Battery Systems. NASA Engineering and Safety Center (NESC) requested NASA Aerospace Flight Battery Systems Working Group to develop a set of guideline requirements document for Binding Procurement Contracts.

  18. In situ determination of V(V) by diffusive gradients in thin films and inductively coupled plasma mass spectrometry techniques using amberlite IRA-410 resin as a binding layer.

    Science.gov (United States)

    Luko, Karen Silva; Menegário, Amauri Antonio; Suárez, Carlos Alfredo; Tafurt-Cardona, Makenly; Pedrobom, Jorge Henrique; Rolisola, Ana Marta Cavinato Marchini; Sulato, Everton Tiago; Kiang, Chang Hung

    2017-01-15

    Amberlite IRA-410 anionic exchange resin was evaluated as the binding layer for sampling V(V) by using Diffusive Gradients in Thin Films (DGT). V(V) was determined by inductively coupled plasma mass spectrometry (ICP-MS). Mass vs. time DGT deployments (ionic strength = 0.03 mol L(-1) NaNO3, pH = 5.6 and T = 23.5 ± 0.5 °C) was characterized by excellent linear relationship (R(2) = 0.9993) and a significant retention of V(V) by the binding layer. An exchange capacity of at least 40 μg V g(-1) resin was achieved for the proposed binding layer. The diffusion coefficient obtained (7.13 ± 0.6 10(-6) cm(2) s(-1)) agrees with the literature. The accumulation rate of V(V) was not significantly affected by ionic strength of solutions up to 0.03 mol L(-1) and for the entire studied pH range (from 3 to 9). Furthermore, when comparing the concentrations obtained using IRA-410-DGT and those obtained by direct measurement of the solution concentrations, the proposed approach provided a reduction of the (35)Cl(16)O interference on V(V) determination by ICP-MS. Determination of V in normal mode (without collision cell) in solutions containing analyte:Cl(-) concentration ratio up to 1:500,000 was not affected by interference of (35)Cl(16)O(+) polyatomic ion even when normal mode ICP-MS was used. Potential interfering ions on sampling V(V) by DGT (PO4(3-) and SO4(2-)) showed no significant effects on the accumulation rate of V(V). Laboratory tests performed using synthetic samples, natural freshwater and acid drainage water showed an excellent performance (recoveries from 93% to 110%). For in situ deployment, measurements of V(V) by the proposed approach was not significantly different (95.5%) from the value of dissolved V concentration.

  19. Microheterogeneity of transthyretin in serum and ascitic fluid of ovarian cancer patients

    Directory of Open Access Journals (Sweden)

    Mustea Alexander

    2005-10-01

    Full Text Available Abstract Background Transthyretin (TTR, a traditional biomarker for nutritional and inflammatory status exists in different molecular variants of yet unknown importance. A truncated form of TTR has recently been described to be part of a set of biomarkers for the diagnosis of ovarian cancer. The main aim of the study was therefore to characterize differences in microheterogeneity between ascitic fluid and plasma of women affected with ovarian cancer and to evaluate the tumor site as the possible source of TTR. Methods Subjects were 48 women with primary invasive epithelial ovarian cancer or recurrent ovarian carcinoma. The control group consisted of 20 postmenopausal women. TTR and retinol-binding protein (RBP levels were measured by enzyme-linked immunoassay (ELISA and C-reactive protein (CRP levels by a high-sensitivity latex particle turbidimetric assay. The molecular heterogeneity of TTR was analysed using immunoprecipitation and matrix-associated laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS. Presence of TTR in tumor tissue was determined with indirect peroxidase immunostaining. Results TTR and RBP (μg/ml levels in serum were 148.5 ± 96.7 and 22.5 ± 14.8 in affected women compared to 363.3 ± 105.5 and 55.8 ± 9.3 in healthy postmenopausal women (p 40 mg/ml (p = 0.08 for TTR; p Conclusion The severity of the cancer associated catabolism as well as the inflammation status affect serum TTR and RBP levels. Neither TTR nor its truncated form originates from tumor tissue and its occurrence in ascites may well reflect the filtration from blood into ascitic fluid.

  20. Toward the discovery of functional transthyretin amyloid inhibitors: application of virtual screening methods.

    Science.gov (United States)

    Simões, Carlos J V; Mukherjee, Trishna; Brito, Rui M M; Jackson, Richard M

    2010-10-25

    Inhibition of amyloid fibril formation by stabilization of the native form of the protein transthyretin (TTR) is a viable approach for the treatment of familial amyloid polyneuropathy that has been gaining momentum in the field of amyloid research. The TTR stabilizer molecules discovered to date have shown efficacy at inhibiting fibrilization in vitro but display impairing issues of solubility, affinity for TTR in the blood plasma and/or adverse effects. In this study we present a benchmark of four protein- and ligand-based virtual screening (VS) methods for identifying novel TTR stabilizers: (i) two-dimensional (2D) similarity searches with chemical hashed, pharmacophore, and UNITY fingerprints, (ii) 3D searches based on shape, chemical, and electrostatic similarity, (iii) LigMatch, a new ligand-based method which uses multiple templates and combines 3D geometric hashing with a 2D preselection process, and (iv) molecular docking to consensus X-ray crystal structures of TTR. We illustrate the potential of the best-performing VS protocols to retrieve promising new leads by ranking a tailored library of 2.3 million commercially available compounds. Our predictions show that the top-scoring molecules possess distinctive features from the known TTR binders, holding better solubility, fraction of halogen atoms, and binding affinity profiles. To the best of our knowledge, this is the first attempt to rationalize the utilization of a large battery of in silico screening techniques toward the identification of a new generation of TTR amyloid inhibitors.

  1. Melissa officinalis essential oil reduces plasma triglycerides in human apolipoprotein E2 transgenic mice by inhibiting sterol regulatory element-binding protein-1c-dependent fatty acid synthesis.

    Science.gov (United States)

    Jun, Hee-Jin; Lee, Ji Hae; Jia, Yaoyao; Hoang, Minh-Hien; Byun, Hanna; Kim, Kyoung Heon; Lee, Sung-Joon

    2012-03-01

    We investigated the hypolipidemic effects of Melissa officinalis essential oil (MOEO) in human APOE2 transgenic mice and lipid-loaded HepG2 cells. Plasma TG concentrations were significantly less in APOE2 mice orally administered MOEO (12.5 μg/d) for 2 wk than in the vehicle-treated group. Cellular TG and cholesterol concentrations were also significantly decreased in a dose- (400 and 800 mg/L) and time- (12 and 24 h) dependent manner in HepG2 cells stimulated with MOEO compared with controls. Mouse hepatic transcriptome analysis suggested MOEO feeding altered several lipid metabolic pathways, including bile acid and cholesterol synthesis and fatty acid metabolism. In HepG2 cells, the rate of fatty acid oxidation, as assessed using [1-(14)C]palmitate, was unaltered; however, the rate of fatty acid synthesis quantified with [1-(14)C]acetate was significantly reduced by treatment with 400 and 800 mg/L MOEO compared with untreated controls. This reduction was due to the decreased expression of SREBP-1c and its responsive genes in fatty acid synthesis, including FAS, SCD1, and ACC1. Subsequent chromatin immunoprecipitation analysis further demonstrated that the binding of p300/CBP-associated factor, a coactivator of SREBP-1c, and histone H3 lysine 14 acetylation at the FAS, SCD1, and ACC1 promoters were significantly reduced in the livers of APOE2 mice and HepG2 cells treated with MOEO compared with their controls. Additionally, MOEO stimulation in HepG2 cells induced bile acid synthesis and reduced the nuclear form of SREBP-2, a key transcription factor in hepatic cholesterol synthesis. These findings suggest that the intake of phytochemicals with pleasant scent could have beneficial metabolic effects.

  2. A Thr94Ala mutation in human liver fatty acid-binding protein contributes to reduced hepatic glycogenolysis and blunted elevation of plasma glucose levels in lipid-exposed subjects.

    Science.gov (United States)

    Weickert, Martin O; Loeffelholz, Christian V; Roden, Michael; Chandramouli, Visvanathan; Brehm, Attila; Nowotny, Peter; Osterhoff, Martin A; Isken, Frank; Spranger, Jochen; Landau, Bernard R; Pfeiffer, Andreas F H; Möhlig, Matthias

    2007-10-01

    Liver fatty acid-binding protein (L-FABP) is a highly conserved key factor in lipid metabolism. Amino acid replacements in L-FABP might alter its function and thereby affect glucose metabolism in lipid-exposed subjects, as indicated by studies in L-FABP knockout mice. Amino acid replacements in L-FABP were investigated in a cohort of 1,453 Caucasian subjects. Endogenous glucose production (EGP), gluconeogenesis, and glycogenolysis were measured in healthy carriers of the only common Thr(94)-to-Ala amino acid replacement (Ala/Ala(94)) vs. age-, sex-, and BMI-matched wild-type (Thr/Thr(94)) controls at baseline and after 320-min lipid/heparin-somatostatin-insulin-glucagon clamps (n = 18). Whole body glucose disposal was further investigated (subset; n = 13) using euglycemic-hyperinsulinemic clamps without and with lipid/heparin infusion. In the entire cohort, the only common Ala/Ala(94) mutation was significantly associated with reduced body weight, which is in agreement with a previous report. In lipid-exposed, individually matched subjects there was a genotype vs. lipid-treatment interaction for EGP (P = 0.009) driven mainly by reduced glycogenolysis in Ala/Ala(94) carriers (0.46 +/- 0.05 vs. 0.59 +/- 0.05 mgxkg(-1)xmin(-1), P = 0.013). The lipid-induced elevation of plasma glucose levels was smaller in Ala/Ala(94) carriers compared with wild types (P glycogenolysis and less severe hyperglycemia in lipid-exposed humans and was further associated with reduced body weight in a large cohort. Data clearly show that investigation of L-FABP phenotypes in the basal overnight-fasted state yielded incomplete information, and a challenge test was essential to detect phenotypical differences in glucose metabolism between L-FABP genotypes.

  3. Determination of MBL Gene by Using PCR-RFLP and the Study on the Association of Plasma Concentrations of Mannose Binding Protein in Healthy Individuals of Han and Mongolian Population

    Institute of Scientific and Technical Information of China (English)

    贾天军; 李萍; 张庶民; 刘洁; 赵铁军; 张进顺; 雷殿良; 翟登高; 李河民

    2004-01-01

    The aim of this study was to analyze the point mutation of the exon 1 at codon 54 of the mannose ( or mannan)-binding lectin (MBL) gene in healthy individuals of Chinese Hans and Mongolian population, and to find out any association between the plasma levels of MBL and the gene mutation frequency in beth groups of individuals. Blood samples were collected randomly from 56 healthy individuals of Chinese Hans and 37 Mongolian. The detection of the point mutations of the MBL gene was performed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and detections for plasma levels of MBL were determined by using MBL ELISA kits. A MBL PCR method of assay was established with high specificity, and good reproducibility. By optimizing the PCR condition, the optimal annealing temperature was 55℃, and the lowest detec-tion limit was 160 pg. No bands were found in non-specificity samples (HAV, HBV, HCV and TB), and the sequences of PCR products were the same as the expected ones. Also a MBL PCR-RFIP was established. Upon electrophoresis of the digested products in 3% agarose gel, there were 3 patterns: in which 2 bands correspond to molecule weight 232 bp and 93 bp;1 band, corresponds to molecule weight 325 bp and 3 bands correspond to molecule weight 325 bp, 232 bp and 93 bp, respectively. Three bands of 325 bp, 232 bp and 93 bp of point mutations were found at cedon 54 of MBL ceding gene. Frequencies in healthy Han and Mongolian population were 0. 2321 and 0. 1757 respectively. The average plasma MBL concentration was 1998.750μg/L, with SD of 1505. 152 in 56 healthy Han population and 2525.676μg/L, with SD of 1955. 188 in 37 Mongolian. A negative correlation between MBL concentration and gene mutation frequency was found in healthy Han population. Frequency of point mutation was 1.00 when the MBL concentrations were below 100μg/L; frequency of point mutation was0.4524 when the concentration was 100μg/L to 1000μg/L; and the frequency of point

  4. Photoaffinity Labeling of Plasma Proteins

    Directory of Open Access Journals (Sweden)

    Masaki Otagiri

    2013-11-01

    Full Text Available Photoaffinity labeling is a powerful technique for identifying a target protein. A high degree of labeling specificity can be achieved with this method in comparison to chemical labeling. Human serum albumin (HSA and α1-acid glycoprotein (AGP are two plasma proteins that bind a variety of endogenous and exogenous substances. The ligand binding mechanism of these two proteins is complex. Fatty acids, which are known to be transported in plasma by HSA, cause conformational changes and participate in allosteric ligand binding to HSA. HSA undergoes an N-B transition, a conformational change at alkaline pH, that has been reported to result in increased ligand binding. Attempts have been made to investigate the impact of fatty acids and the N-B transition on ligand binding in HSA using ketoprofen and flunitrazepam as photolabeling agents. Meanwhile, plasma AGP is a mixture of genetic variants of the protein. The photolabeling of AGP with flunitrazepam has been utilized to shed light on the topology of the protein ligand binding site. Furthermore, a review of photoaffinity labeling performed on other major plasma proteins will also be discussed. Using a photoreactive natural ligand as a photolabeling agent to identify target protein in the plasma would reduce non-specific labeling.

  5. Plasma turbulence

    Energy Technology Data Exchange (ETDEWEB)

    Horton, W. [Univ. of Texas, Austin, TX (United States). Inst. for Fusion Studies; Hu, G. [Globalstar LP, San Jose, CA (United States)

    1998-07-01

    The origin of plasma turbulence from currents and spatial gradients in plasmas is described and shown to lead to the dominant transport mechanism in many plasma regimes. A wide variety of turbulent transport mechanism exists in plasmas. In this survey the authors summarize some of the universally observed plasma transport rates.

  6. Plasma harmonics

    CERN Document Server

    Ganeev, Rashid A

    2014-01-01

    Preface; Why plasma harmonics? A very brief introduction Early stage of plasma harmonic studies - hopes and frustrations New developments in plasma harmonics studies: first successes Improvements of plasma harmonics; Theoretical basics of plasma harmonics; Basics of HHG Harmonic generation in fullerenes using few-cycle pulsesVarious approaches for description of observed peculiarities of resonant enhancement of a single harmonic in laser plasmaTwo-colour pump resonance-induced enhancement of odd and even harmonics from a tin plasmaCalculations of single harmonic generation from Mn plasma;Low-o

  7. The binding site for regulatory 14-3-3 protein in plant plasma membrane H+-ATPase: Involvement of a region promoting phosphorylation-independent interaction in addition to the phosphorylation-dependent C-terminal end

    DEFF Research Database (Denmark)

    Fuglsang, Anja T; Borch, Jonas; Bych, Katrine

    2003-01-01

    ) in the extreme C-terminal end of the H+-ATPase interacts with the binding cleft of 14-3-3 protein (Wurtele, M., Jelich-Ottmann, C., Wittinghofer, A., and Oecking, C. (2003) EMBO J. 22, 987-994). We report binding of 14-3-3 protein to a nonphosphorylated peptide representing the 34 C-terminal residues...

  8. Dusty plasmas

    Energy Technology Data Exchange (ETDEWEB)

    Fortov, Vladimir E; Khrapak, Aleksei G; Molotkov, Vladimir I; Petrov, Oleg F [Institute for High Energy Densities, Associated Institute for High Temperatures, Russian Academy of Sciences, Moscow (Russian Federation); Khrapak, Sergei A [Max-Planck-Institut fur Extraterrestrische Physik, Garching (Germany)

    2004-05-31

    The properties of dusty plasmas - low-temperature plasmas containing charged macroparticles - are considered. The most important elementary processes in dusty plasmas and the forces acting on dust particles are investigated. The results of experimental and theoretical investigations of different states of strongly nonideal dusty plasmas - crystal-like, liquid-like, gas-like - are summarized. Waves and oscillations in dusty plasmas, as well as their damping and instability mechanisms, are studied. Some results on dusty plasma investigated under microgravity conditions are presented. New directions of experimental research and potential applications of dusty plasmas are discussed. (reviews of topical problems)

  9. Use of a column-switching high-performance liquid chromatography method to assess the presence of specific binding of (R)- and (S)-[{sup 11}C]rolipram and their labeled metabolites to the phosphodiesterase-4 enzyme in rat plasma and tissues

    Energy Technology Data Exchange (ETDEWEB)

    Kenk, Miran; Greene, Michael [Cardiovascular PET Molecular Imaging Program, Division of Cardiology, University of Ottawa Heart Institute, Ottawa, Ontario, K1Y 4W7 (Canada); Department of Cellular and Molecular Medicine, Faculty of Medicine, University of Ottawa, Ottawa, Ontario, K1H 8M5 (Canada); Lortie, Mireille; Kemp, Robert A. de [Cardiovascular PET Molecular Imaging Program, Division of Cardiology, University of Ottawa Heart Institute, Ottawa, Ontario, K1Y 4W7 (Canada); Beanlands, Rob S. [Cardiovascular PET Molecular Imaging Program, Division of Cardiology, University of Ottawa Heart Institute, Ottawa, Ontario, K1Y 4W7 (Canada); Department of Cellular and Molecular Medicine, Faculty of Medicine, University of Ottawa, Ottawa, Ontario, K1H 8M5 (Canada); DaSilva, Jean N. [Cardiovascular PET Molecular Imaging Program, Division of Cardiology, University of Ottawa Heart Institute, Ottawa, Ontario, K1Y 4W7 (Canada); Department of Cellular and Molecular Medicine, Faculty of Medicine, University of Ottawa, Ottawa, Ontario, K1H 8M5 (Canada)], E-mail: jdasilva@ottawaheart.ca

    2008-05-15

    Introduction: To complement recent studies using the high-affinity {sup 11}C-labeled phosphodiesterase-4 (PDE4) inhibitor (R)-rolipram and the less active enantiomer (S)-[{sup 11}C]rolipram for in vivo quantification of PDE4 levels, we evaluated the presence of radiolabeled metabolites and their potential binding to PDE4 in the rat plasma, brain, heart, pancreas, skeletal muscle and brown adipose tissue. Methods: A reverse-phase capture and analytical HPLC column-switch method was used to detect (R)-[{sup 11}C]rolipram, (S)-[{sup 11}C]rolipram and their radiolabeled metabolites in rat plasma and tissue extracts. The relative proportion of PDE4-specific binding of the radiotracers and their labeled metabolites was analyzed following co-injections with a saturating dose of unlabeled (R)-rolipram at 45 min post-tracer injection in tissue extracts. Results: Radiolabeled metabolites were found in the plasma (72-75% of total radioactive signal), and in the heart, skeletal muscle, pancreas and brown adipose tissue (44-52%), but not in the brain. In comparison to polar labeled metabolites, the proportion of unchanged (R)-[{sup 11}C]rolipram was reduced in PDE4-rich organs by co-injection of unlabeled (R)-rolipram. Conversely, no changes were obtained in brown adipose tissue, or with (S)-[{sup 11}C]rolipram, suggesting that radiolabeled metabolites of (R)-[{sup 11}C]rolipram display no specific binding to PDE4. Conclusions: Radiolabeled hydrophilic metabolites are unlikely to compete with (R)-[{sup 11}C]rolipram for PDE4-specific retention. However, due to the high proportion of the radioactive metabolites in the total radioactive signal, any kinetic modeling calculations in the peripheral tissues will need to take into account the presence of labeled metabolites.

  10. Retinoid-binding proteins: similar protein architectures bind similar ligands via completely different ways.

    Directory of Open Access Journals (Sweden)

    Yu-Ru Zhang

    Full Text Available BACKGROUND: Retinoids are a class of compounds that are chemically related to vitamin A, which is an essential nutrient that plays a key role in vision, cell growth and differentiation. In vivo, retinoids must bind with specific proteins to perform their necessary functions. Plasma retinol-binding protein (RBP and epididymal retinoic acid binding protein (ERABP carry retinoids in bodily fluids, while cellular retinol-binding proteins (CRBPs and cellular retinoic acid-binding proteins (CRABPs carry retinoids within cells. Interestingly, although all of these transport proteins possess similar structures, the modes of binding for the different retinoid ligands with their carrier proteins are different. METHODOLOGY/PRINCIPAL FINDINGS: In this work, we analyzed the various retinoid transport mechanisms using structure and sequence comparisons, binding site analyses and molecular dynamics simulations. Our results show that in the same family of proteins and subcellular location, the orientation of a retinoid molecule within a binding protein is same, whereas when different families of proteins are considered, the orientation of the bound retinoid is completely different. In addition, none of the amino acid residues involved in ligand binding is conserved between the transport proteins. However, for each specific binding protein, the amino acids involved in the ligand binding are conserved. The results of this study allow us to propose a possible transport model for retinoids. CONCLUSIONS/SIGNIFICANCE: Our results reveal the differences in the binding modes between the different retinoid-binding proteins.

  11. Imidazole binding to human serum albumin.

    Science.gov (United States)

    Rodrigo, M C; Ceballos, A; Mariño, E; Cachaza, J M; Domínguez-Gil, A; Kuemmerle, H P

    1988-06-01

    Imidazole is a substance released by the organism when a new salicylate derivative, imidazole salicylate is administered. A study was made of the binding of imidazole to human serum albumin by an in vitro assay employing an ultrafiltration technique. For the concentration range that imidazole was found in plasma following administration of the drug to healthy volunteers, the mean binding percentages were: 12.1 +/- 1.8 and 19.7 +/- 3.1 at 37 degrees C and 25 degrees C, respectively. The results obtained in the study follow a model entailing three equal and independent binding sites of imidazole to serum albumin and the values of the corresponding constants were determined. Apparently, the presence in the plasma samples of sodium salicylate at a concentration of 100 micrograms/ml does not affect the binding of imidazole to human serum albumin.

  12. Is Lutein a Physiologically Important Ligand for Transthyretin in Humans?

    Energy Technology Data Exchange (ETDEWEB)

    Liwei Chen

    2003-05-31

    Lutein and zeaxanthin are the only carotenoids accumulated in the macula of the human retina and are known as the macular pigments (MP). These pigments account for the yellow color of the macula and appear to play an important role in protecting against age-related macular degeneration (AMD). The uptake of lutein and zeaxanthin in human eyes is remarkably specific. It is likely that specific transport or binding proteins are involved. The objective is to determine whether transthyretin (TTR) is a transport protein in human plasma and could thus deliver lutein from the blood to the retina. In this study, they used a biosynthetic {sup 13}C-lutein tracer and gas chromatography-combustion interfaced-isotope ratio mass spectrometry (GCC-IRMS) to gain the requisite sensitivity to detect the minute amounts of lutein expected as a physiological ligand for human transthyretin. The biosynthetic {sup 13}C-labeled lutein tracer was purified from algae. Healthy women (n = 4) each ingested 1 mg of {sup 13}C-labeled lutein daily for 3 days and a blood sample was collected 24 hours after the final dose. Plasma TTR was isolated by retinol-binding protein (RBP)-sepharose affinity chromatography and extracted with chloroform. The {sup 13}C/{sup 12}C ratio in the TTR extract was measured by GCC-IRMS. There was no {sup 13}C-lutein enrichment in the pure TTR extract. This result indicated that lutein is not associated with TTR in human plasma after ingestion in physiological amounts. Some hydrophobic compounds with yellow color may bind to human TTR in the plasma. However, this association needs to be further proved by showing specificity. The study provides a new approach for carotenoid-binding protein studies using a stable isotope tracer method combined with the high precision of GCC-IRMS. The mechanism of selective transport, uptake, and accumulation of lutein in human macula remain to be determined.

  13. Is Lutein a Physiologically Important Ligand for Transthyretin in Humans?

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Liwei [Iowa State Univ., Ames, IA (United States)

    2003-01-01

    Lutein and zeaxanthin are the only carotenoids accumulated in the macula of the human retina and are known as the macular pigments (MP). These pigments account for the yellow color of the macula and appear to play an important role in protecting against age-related macular degeneration (AMD). The uptake of lutein and zeaxanthin in human eyes is remarkably specific. It is likely that specific transport or binding proteins are involved. The objective is to determine whether transthyretin (TTR) is a transport protein in human plasma and could thus deliver lutein from the blood to the retina. In this study, they used a biosynthetic 13C-lutein tracer and gas chromatography-combustion interfaced-isotope ratio mass spectrometry (GCC-IRMS) to gain the requisite sensitivity to detect the minute amounts of lutein expected as a physiological ligand for human transthyretin. The biosynthetic 13C-labeled lutein tracer was purified from algae. Healthy women (n = 4) each ingested 1 mg of 13C-labeled lutein daily for 3 days and a blood sample was collected 24 hours after the final dose. Plasma TTR was isolated by retinol-binding protein (RBP)-sepharose affinity chromatography and extracted with chloroform. The 13C/12C ratio in the TTR extract was measured by GCC-IRMS. There was no 13C-lutein enrichment in the pure TTR extract. This result indicated that lutein is not associated with TTR in human plasma after ingestion in physiological amounts. Some hydrophobic compounds with yellow color may bind to human TTR in the plasma. However, this association needs to be further proved by showing specificity. The study provides a new approach for carotenoid-binding protein studies using a stable isotope tracer method combined with the high precision of GCC-IRMS. The mechanism of selective transport, uptake, and accumulation of lutein in human macula remain to be determined.

  14. Determination of plasma protein binding rate both in Polygonum orientale L.extract and compound Hongcao freeze-dried powder for injection by equilibrium dialysis%平衡透析法研究荭草及其制剂的人血浆蛋白结合率

    Institute of Scientific and Technical Information of China (English)

    黄勇; 陈慧; 何峰; 张治蓉; 王永林

    2012-01-01

    OBJECTIVE To determine and compare the plasma protein binding rate of two kinds of compounds in the extraction of Polygonum orientate and compound Hongcao freeze-dned powder for injection by equilibrium dialysis method. METHODS Equilibrium dialysis method combined with liquid chromatography-tandem mass spectrometry (UPLC-ESI-MS/MS) was employed to determine the protein binding rate of protocatechuic acid, isoorientin with human plasma protein both in the extrac- tion of Polygonum orientate and compound Hongcao freeze-dried powder and the binding characteristics were also calculated. RESULTS The results showed medium binding power of protocatechuic acid and high binding power of isoorientin to plasma protein, the powers of the two compounds with the plasma protein were independent on the area of the concentration which was determinate. In the compound Hongcao freeze-dried powder, the related binding characteristics of isoorientin had somewhat increase; and compared with the extraction of Polygonum orientate , there was also some difference. CONCLUSION The protein binding rates of the compounds in the Polygonum orientate were independent on the area of the concentration which was determinate, but there was some change when the herb was composed to compound preparation.%目的:比较荭草提取物及其复方制剂中两类成分的人血浆蛋白结合率.方法:以超高效液相色谱—质谱联用为检测手段,结合平衡透析法考察单味荭草提取物及注射用复方荭草在人血浆中的蛋白结合率并计算相关参数.结果:在所研究浓度范围内,原儿茶酸与血浆蛋白具有中等强度结合,异荭草素与血浆蛋白结合力较强.经统计分析,两者与血浆蛋白的结合能力在考察浓度范围无显著差异;其中异荭草素指标在复方中的血浆蛋白结合率有所升高,复方中其他的血浆蛋白结合动力学参数相对单味荭草也有一定变化.结论:荭草中化学成分的血浆蛋白结合

  15. Plasma waves

    CERN Document Server

    Swanson, DG

    1989-01-01

    Plasma Waves discusses the basic development and equations for the many aspects of plasma waves. The book is organized into two major parts, examining both linear and nonlinear plasma waves in the eight chapters it encompasses. After briefly discussing the properties and applications of plasma wave, the book goes on examining the wave types in a cold, magnetized plasma and the general forms of the dispersion relation that characterize the waves and label the various types of solutions. Chapters 3 and 4 analyze the acoustic phenomena through the fluid model of plasma and the kinetic effects. Th

  16. Plasma astrophysics

    CERN Document Server

    Kaplan, S A; ter Haar, D

    2013-01-01

    Plasma Astrophysics is a translation from the Russian language; the topics discussed are based on lectures given by V.N. Tsytovich at several universities. The book describes the physics of the various phenomena and their mathematical formulation connected with plasma astrophysics. This book also explains the theory of the interaction of fast particles plasma, their radiation activities, as well as the plasma behavior when exposed to a very strong magnetic field. The text describes the nature of collective plasma processes and of plasma turbulence. One author explains the method of elementary

  17. Tafamidis for transthyretin amyloidosis.

    Science.gov (United States)

    de Lartigue, J

    2012-05-01

    Tafamidis meglumine (Vyndaqel®, Pfizer) is a novel, first-in-class drug for the treatment of transthyretin familial amyloid polyneuropathy (TTR-FAP), a rare neurodegenerative disorder characterized by progressive sensory, motor and autonomic impairment that is ultimately fatal. Pathogenic mutations in the transthyretin (TTR) protein lead to destabilization of its tetrameric structure and subsequent formation of amyloid aggregates. Tafamidis is a small-molecule inhibitor that binds selectively to TTR in human plasma and kinetically stabilizes the tetrameric structure of both wild-type TTR and a number of different mutants. Clinical trials indicate that tafamidis slows disease progression in patients with TTR-FAP and reduces the burden of disease, demonstrating improvement in small and large nerve fiber function, modified body mass index and lower extremity neurological examination. Tafamidis has been granted marketing authorization by the European Commission for the treatment of TTR-FAP and the U.S. Food and Drug Administration is currently reviewing this drug for the same indication.

  18. Changes and Clinical Significance of Plasma Endotoxin and Serum Mannan-binding Lectin Levels in Patients with Hepatitis C%慢性丙型肝炎患者血浆内毒素和血清MBL变化及临床意义

    Institute of Scientific and Technical Information of China (English)

    王洪

    2011-01-01

    Objective To explore the changes and significance of plasma endotoxin and serum mannan-binding lectin( MBL) levels in patients with hepatitis C. Methods Plasma endotoxin (with limulus test) serum MBL (with ELISA) levels were measured in 31 patients with hepatitis C and 35 normal controls. Results plasma endotoxin and MBL levels were extremely higher than those in controls (P<0. 01) . Plasma endotoxin level was positive correlations with MBL levels (r = 0.4784, P <0.01) . Conclusion Detection of plasma endotoxin and MBL levels domonstrate that HCV infection resulted in chronic pathologic process suggesting it possibly play a role in the development of fibrosis in hepatitis C virus-infected patients.%目的:探讨了慢性丙型肝炎患者血浆内毒素和血清甘露聚糖结合凝集素(MBL)水平的变化及意义.方法:应用鲎试验和酶联法对31例慢性丙型肝炎患者进行了血浆内毒素和血清MBL测定,并与35名正常健康人作比较.结果:慢性丙型肝炎患者血浆内毒素和血清MBL水平均非常显著地高于正常人水平(P<0.01),且内毒素水平与MBL呈正相关(r = 0.4784,P<0.01).结论:血浆内毒素和血清MBL可反映HCV感染导致慢性病理状态,它可能也参与肝脏纤维化的过程.

  19. Localization of the ATP-binding cassette (ABC) transport proteins PfMRP1, PfMRP2, and PfMDR5 at the Plasmodium falciparum plasma membrane.

    NARCIS (Netherlands)

    Kavishe, R.A.; Heuvel, J.M.W. van den; Vegte-Bolmer, M. van de; Luty, A.J.; Russel, F.G.M.; Koenderink, J.B.

    2009-01-01

    BACKGROUND: The spread of drug resistance has been a major obstacle to the control of malaria. The mechanisms underlying drug resistance in malaria seem to be complex and multigenic. The current literature on multiple drug resistance against anti-malarials has documented PfMDR1, an ATP-binding casse

  20. Plasma protein haptoglobin modulates renal iron loading

    DEFF Research Database (Denmark)

    Fagoonee, Sharmila; Gburek, Jakub; Hirsch, Emilio

    2005-01-01

    Haptoglobin is the plasma protein with the highest binding affinity for hemoglobin. The strength of hemoglobin binding and the existence of a specific receptor for the haptoglobin-hemoglobin complex in the monocyte/macrophage system clearly suggest that haptoglobin may have a crucial role in heme...

  1. Analyzing binding data.

    Science.gov (United States)

    Motulsky, Harvey J; Neubig, Richard R

    2010-07-01

    Measuring the rate and extent of radioligand binding provides information on the number of binding sites, and their affinity and accessibility of these binding sites for various drugs. This unit explains how to design and analyze such experiments.

  2. Factor XII binding to endothelial cells depends on caveolae

    DEFF Research Database (Denmark)

    Schousboe, Inger; Thomsen, Peter; van Deurs, Bo

    2004-01-01

    to human umbilical vein endothelial cells (HUVEC) has never been shown to be localized to these specialized membrane structures. Using microscopical techniques, we here report that FXII binds to specific patches of the HUVEC plasma membrane with a high density of caveolae. Further investigations of FXII...... lipid rafts. Accordingly, cholesterol-depleted cells were found to bind significantly reduced amounts of FXII. These observations, combined with the presence of a minority of u-PAR in caveolae concomitant with FXII binding, indicate that FXII binding to u-PAR may be secondary and depends upon...... the structural elements within caveolae. Thus, FXII binding to HUVEC depends on intact caveolae on the cellular surface....

  3. Plasma Antenna

    OpenAIRE

    N M Vijay

    2014-01-01

    The fundamental base of plasma antenna is the use of an ionized medium as a conductor. The plasma antenna is a radiofrequency antenna formed by a plasma columns, Filaments or sheets, which are excited by a surface wave. The relevance of this device is how rapidly it can be turned on and off, only applying an electrical pulse. Besides its wide carrier frequency, the great directivity and controllable antenna shape. Otherwise a disadvantage is that it needs energy to be ionized....

  4. Plasma physics

    CERN Document Server

    Drummond, James E

    2013-01-01

    A historic snapshot of the field of plasma physics, this fifty-year-old volume offers an edited collection of papers by pioneering experts in the field. In addition to assisting students in their understanding of the foundations of classical plasma physics, it provides a source of historic context for modern physicists. Highly successful upon its initial publication, this book was the standard text on plasma physics throughout the 1960s and 70s.Hailed by Science magazine as a ""well executed venture,"" the three-part treatment ranges from basic plasma theory to magnetohydrodynamics and microwa

  5. Identification of fibrin clot-bound plasma proteins

    NARCIS (Netherlands)

    S. Talens (Simone); F.W.G. Leebeek (Frank); J.A.A. Demmers (Jeroen); D.C. Rijken (Dingeman)

    2012-01-01

    textabstractSeveral proteins are known to bind to a fibrin network and to change clot properties or function. In this study we aimed to get an overview of fibrin clot-bound plasma proteins. A plasma clot was formed by adding thrombin, CaCl2 and aprotinin to citrated platelet-poor plasma and unbound

  6. Binding Affinity and Capacity for the Uremic Toxin Indoxyl Sulfate

    Directory of Open Access Journals (Sweden)

    Eric Devine

    2014-01-01

    Full Text Available Protein binding prevents uremic toxins from removal by conventional extracorporeal therapies leading to accumulation in maintenance dialysis patients. Weakening of the protein binding may enhance the dialytic elimination of these toxins. In ultrafiltration and equilibrium dialysis experiments, different measures to modify the plasma binding affinity and capacity were tested: (i, increasing the sodium chloride (NaCl concentration to achieve a higher ionic strength; (ii, increasing the temperature; and (iii, dilution. The effects on the dissociation constant KD and the protein bound fraction of the prototypical uremic toxin indoxyl sulfate (IS in plasma of healthy and uremic individuals were studied. Binding of IS corresponded to one site binding in normal plasma. KD increased linearly with the NaCl concentration between 0.15 (KD = 13.2 ± 3.7 µM and 0.75 M (KD = 56.2 ± 2.0 µM. Plasma dilution further reduced the protein bound toxin fraction by lowering the protein binding capacity of the plasma. Higher temperatures also decreased the protein bound fraction of IS in human plasma. Increasing the NaCl concentration was effective to weaken the binding of IS also in uremic plasma: the protein bound fraction decreased from 89% ± 3% to 81% ± 3% at 0.15 and 0.75 M NaCl, respectively. Dilution and increasing the ionic strength and temperature enhance the free fraction of IS allowing better removal of the substance during dialysis. Applied during clinical dialysis, this may have beneficial effects on the long-term outcome of maintenance dialysis patients.

  7. Skjaldar þáttr Danakonungs

    DEFF Research Database (Denmark)

    Lavender, Philip Thomas

    2016-01-01

    In this article I discuss a reference in Illuga saga Gríðarfóstra to a Skjaldar þáttur ok Hermanns. Previous commentators have said that this reference is either made up or alludes to a now lost saga. I show that the reference to the saga is not a fabrication: the narrative of Skjöldr and Hermann...... exists in several different forms present in post-medieval manuscripts. A diplomatic edition of one of the earliest texts is provided here....

  8. Relationship between Plasma Retinol-binding Protein 4 and Intima-media Thickness of Common Iliac Artery in Newly Diagnosed Type 2 Diabetics%新诊T2DM患者血浆RBP4与髂动脉内中膜厚度的关系研究

    Institute of Scientific and Technical Information of China (English)

    钟慧; 漆辉洲; 游咏; 陈雯; 冯聚玲; 谢娟; 李熠

    2014-01-01

    Objective To evaluate the relationship between the concentration of plasma retinol-binding protein 4 (RBP4) and intima-media thickness of common iliac artery (CIA-IMT) in newly diagnosed type 2 diabetic (T2DM) patients. Methods There were 180 newly diagnosed T2DM patients in study. Their plasma retinol-binding protein 4 level and other clinic index were tested. The patients were divided into three groups based on the concentrations of RBP4 to compare their CIA-IMT. The relevance of RBP4 and other parameters were analyzed by Pearson correlation analysis. Results CIA-IMT of RBP4 high tertile group was significantly thicker than other groups ( P<0.05). It is showed by Pearson correlation analysis that the concentration of RBP4 in T2DM patients is positively correlated with BMI, FBS, PBS, FINS and HOMA-IR.Conclusion CIA-IMT in newly diagnosed T2DM patients is closely correlated with plasma RBP4 levels.%目的:探讨新诊2型糖尿病(T2DM)患者血浆视黄醇结合蛋白4(RBP4)与髂动脉内中膜厚度(CIA-IMT)的关系。方法180例新诊T2DM患者测定血浆RBP4浓度、血糖、糖化血红蛋白(HbA1c)、血脂、胰岛素(FINS)及CIA-IMT,根据RBP4浓度由低到高按三分位法将180例患者分为三组,比较其CIA-IMT;用Pearson相关分析分析RBP4与其他指标的相关性。结果T2DM患者RBP4上三分位组CIA-IMT比其余两组增厚,差异有统计学意义(P<0.05);Pearson相关分析显示T2DM患者血浆RBP4与BMI、FBS、PBS、FINS及HOMA-IR呈正相关。结论新诊T2DM患者的血浆RBP4水平与CIA-IMT密切相关。

  9. Plasma chromograninx

    DEFF Research Database (Denmark)

    Goetze, Jens P; Hilsted, Linda M; Rehfeld, Jens F

    2014-01-01

    Cardiovascular risk assessment remains difficult in elderly patients. We examined whether chromogranin A (CgA) measurement in plasma may be valuable in assessing risk of death in elderly patients with symptoms of heart failure in a primary care setting. A total of 470 patients (mean age 73 years......) were followed for 10 years. For CgA plasma measurement, we used a two-step method including a screening test and a confirmative test with plasma pre-treatment with trypsin. Cox multivariable proportional regression and receiver-operating curve (ROC) analyses were used to assess mortality risk...... of follow-up showed significant additive value of CgA confirm measurements compared with NT-proBNP and clinical variables. CgA measurement in the plasma of elderly patients with symptoms of heart failure can identify those at increased risk of short- and long-term mortality....

  10. Plasma Cleaning

    Science.gov (United States)

    Hintze, Paul E.

    2016-01-01

    NASA's Kennedy Space Center has developed two solvent-free precision cleaning techniques: plasma cleaning and supercritical carbon dioxide (SCCO2), that has equal performance, cost parity, and no environmental liability, as compared to existing solvent cleaning methods.

  11. Plasma confinement

    CERN Document Server

    Hazeltine, R D

    2003-01-01

    Detailed and authoritative, this volume examines the essential physics underlying international research in magnetic confinement fusion. It offers readable, thorough accounts of the fundamental concepts behind methods of confining plasma at or near thermonuclear conditions. Designed for a one- or two-semester graduate-level course in plasma physics, it also represents a valuable reference for professional physicists in controlled fusion and related disciplines.

  12. A single rainbow trout cobalamin-binding protein stands in for three human binders

    DEFF Research Database (Denmark)

    Greibe, Eva; Fedosov, Sergey; Sorensen, Boe S

    2012-01-01

    -binding proteins of the rainbow trout (Oncorhynchus mykiss) and to compare their properties with those of the three human cobalamin-binding proteins. High cobalamin-binding capacity was found in trout stomach (210 pmol/g), roe (400 pmol/g), roe fluid (390 nmol/liter), and plasma (2500 nmol/liter). In all cases...

  13. Membrane binding domains

    OpenAIRE

    Hurley, James H.

    2006-01-01

    Eukaryotic signaling and trafficking proteins are rich in modular domains that bind cell membranes. These binding events are tightly regulated in space and time. The structural, biochemical, and biophysical mechanisms for targeting have been worked out for many families of membrane binding domains. This review takes a comparative view of seven major classes of membrane binding domains, the C1, C2, PH, FYVE, PX, ENTH, and BAR domains. These domains use a combination of specific headgroup inter...

  14. Speciation of manganese binding to biomolecules in pine nuts (Pinus pinea) by two-dimensional liquid chromatography coupled to ultraviolet and inductively coupled plasma mass spectrometry detectors followed by identification by electrospray ionization mass spectrometry.

    Science.gov (United States)

    Arias-Borrego, Ana; García-Barrera, Tamara; Gómez-Ariza, José L

    2008-10-01

    Advances in analytical methodology for speciation of manganese in pine nuts are presented in this work. The approach is based on the use of orthogonal chromatographic systems, namely size-exclusion chromatography (SEC) of the extracts and strong anion exchange (IEC) of the fractions collected by the first column. In both columns, manganese elution is first monitored by a quadrupole inductively coupled plasma mass spectrometry (ICP-MS) instrument equipped with an octopole reaction cell and an ultraviolet (UV) detector. SEC is performed by using two columns covering the molecular weight range from pine nuts samples and the presence of Mn-citrate is confirmed by nanoelectrospray ionization quadrupole time-of-flight mass spectrometry (nESI-QqTOF-MS). In the same fraction, a third Mn-containing peak is detected in the IEC-UV-ICP-MS chromatogram. This peak corresponds to a protein containing Mn that was later submitted to a tryptic digestion and analyzed by nESI-QqTOF. The MS/MS data of a doubly charged peptide are used to obtain the sequence of the protein with the Mascot search engine. The peak turned out to be isocitrate dehydrogenase, a protein commonly associated with Mn.

  15. CD36 binds oxidized low density lipoprotein (LDL) in a mechanism dependent upon fatty acid binding.

    Science.gov (United States)

    Jay, Anthony G; Chen, Alexander N; Paz, Miguel A; Hung, Justin P; Hamilton, James A

    2015-02-20

    The association of unesterified fatty acid (FA) with the scavenger receptor CD36 has been actively researched, with focuses on FA and oxidized low density lipoprotein (oxLDL) uptake. CD36 has been shown to bind FA, but this interaction has been poorly characterized to date. To gain new insights into the physiological relevance of binding of FA to CD36, we characterized FA binding to the ectodomain of CD36 by the biophysical method surface plasmon resonance. Five structurally distinct FAs (saturated, monounsaturated (cis and trans), polyunsaturated, and oxidized) were pulsed across surface plasmon resonance channels, generating association and dissociation binding curves. Except for the oxidized FA HODE, all FAs bound to CD36, with rapid association and dissociation kinetics similar to HSA. Next, to elucidate the role that each FA might play in CD36-mediated oxLDL uptake, we used a fluorescent oxLDL (Dii-oxLDL) live cell assay with confocal microscopy imaging. CD36-mediated uptake in serum-free medium was very low but greatly increased when serum was present. The addition of exogenous FA in serum-free medium increased oxLDL binding and uptake to levels found with serum and affected CD36 plasma membrane distribution. Binding/uptake of oxLDL was dependent upon the FA dose, except for docosahexaenoic acid, which exhibited binding to CD36 but did not activate the uptake of oxLDL. HODE also did not affect oxLDL uptake. High affinity FA binding to CD36 and the effects of each FA on oxLDL uptake have important implications for protein conformation, binding of other ligands, functional properties of CD36, and high plasma FA levels in obesity and type 2 diabetes.

  16. Effect of antihypertensive drugs on plasma adiponection and retinol binding protein 4 in elderly patients with essential hypertension%降压药物对老年高血压患者血浆脂联素和视黄醛结合蛋白4含量的影响

    Institute of Scientific and Technical Information of China (English)

    曹萍; 沈丹; 钟亚; 李睿

    2013-01-01

    目的 观察老年高血压患者应用氨氯地平、培哚普利、缬沙坦3种药物降压治疗前后的血浆脂联素和血浆视黄醛结合蛋白含量的变化. 方法 选择2007年3月至2010年7月年在我院住院的老年高血压患者238例,完成研究1 93例,数字抽签随机分为3组,分别用氨氯地平、培哚普利、缬沙坦治疗,1 2周后观察血压、心率、身高、体质量、腹围、腰围、血脂、血浆脂联素及视黄醛结合蛋白水平等改变. 结果 氨氯地平组、培哚普利组、缬沙坦组治疗后血压均较治疗前下降(均P<0.01);治疗后3组血压比较,差异无统计学意义(均P>0.05).治疗前、后血浆脂联素水平培哚普利组(7.4±1.8)μg/L与(8.3±1.8)μg/L、缬沙坦组(7.5±1.7)μg/L与(8.4±1.9)μg/L较治疗前升高(均P<0.01);治疗后培哚普利组、缬沙坦组血浆脂联素水平高于氨氯地平组(7.6±1.8)μg/L(均P<0.05).治疗后培哚普利组、缬沙坦组血浆视黄醛结合蛋白水平较治疗前降低(P<0.01).氨氯地平组较治疗前血浆视黄醛结合蛋白水平降低,但差异无统计学意义(P>0.05).血浆视黄醛结合蛋白水平治疗后培哚普利组(36.6±14.2) mg/L、缬沙坦组(36.3±14.1)mg/L低于氨氯地平组(42.7±13.8)mg/L,差异有统计学意义(均P<0.01). 结论 培哚普利、缬沙坦能提高老年高血压病患者血浆脂联素水平,降低血浆视黄醛结合蛋白水平,从而起到降压作用以外的心血管保护作用.%Objective To explore the effects of amlodipine,perindopril and valsartan on plasma adiponectin and retinol binding protein 4 in elderly patients with essential hypertension.Methods From March 2007 to July 2010,238 elderly patients with essential hypertension were selected and 193 cases completed this study.Patients were randomly divided into 3 groups:amlodipine group (n=68),perindoprilgroup (n=60) and valsartan group (n=65).Patients in each group were treated with amlodipine

  17. Analyzing radioligand binding data.

    Science.gov (United States)

    Motulsky, Harvey; Neubig, Richard

    2002-08-01

    Radioligand binding experiments are easy to perform, and provide useful data in many fields. They can be used to study receptor regulation, discover new drugs by screening for compounds that compete with high affinity for radioligand binding to a particular receptor, investigate receptor localization in different organs or regions using autoradiography, categorize receptor subtypes, and probe mechanisms of receptor signaling, via measurements of agonist binding and its regulation by ions, nucleotides, and other allosteric modulators. This unit reviews the theory of receptor binding and explains how to analyze experimental data. Since binding data are usually best analyzed using nonlinear regression, this unit also explains the principles of curve fitting with nonlinear regression.

  18. Função renal de pacientes de unidade de terapia intensiva: creatinina plasmática e proteína carreadora do retinol urinário Renal function of intensive care unit patients: plasma creatinine and urinary retinol-binding protein

    Directory of Open Access Journals (Sweden)

    Cristina Satoko Mizoi

    2008-12-01

    ário. CONCLUSÃO: A proteina carreadora do retinol urinário, na prática clínica, pode ser considerada um marcador mais apropriado para o diagnóstico em pacientes com risco de desenvolver uma insuficiência renal aguda, quando comparada com outros marcadores usados rotineiramente. Ademais, a proteina carreadora do retinol urinário apresenta outros aspectos de um bom teste diagnóstico - é um método prático e não-invasivo.OBJECTIVES: The early assessment of renal dysfunction using common markers does not provide either a sensitive or specific indication of renal dysfunction in critically ill patients. More specific and sensitive markers are desirable for the early detection of an initial renal pathophysiological process. Urinary retinol-binding protein could be an alternative method to early evaluation of renal function in these patients. METHODS: This study followed-up 100 critical care patients and assessed their clinical and laboratory variables, including plasma creatinine and urinary retinol-binding ratio, and demographic variables. RESULTS: The sample was characterized by geriatric (63.4±15.6 years, male (68%, being 53% surgical patients. Statistical analysis showed association between plasma creatinine and the following variables: gender (p-0.026, age (p-0.038, use of vasoactive drugs (p-0.003, proteinuria (p-0.025, Acute Physiological Chronic Health Evaluation (APACHE II score (p-0.000, urea (p-0.000, potassium (p-0.003 and estimated creatinine clearance (p-0.000. Urinary retinol-binding protein was correlated with more variables: weight, use of invasive ventilation (p-0.000, use of nonsteroidal antiinflammatory drugs (p-0.018, use of vasoactive drugs (p-0.021, high temperature (>37.5ºC (p-0.005, proteinuria (p-0.000, bilirubinuria (p-0.004, urinary flow (p-0.019, minimal diastolic pressure (p-0.032, minimal systolic pressure (p-0.029, APACHE II (p-0.000, creatinine (p-0.001, urea (p-0.001, estimated creatinine clearance (p-0.000. Urinary retinol-binding protein

  19. Mediaeval manuscript bindings

    Directory of Open Access Journals (Sweden)

    Jedert Vodopivec

    1999-01-01

    Full Text Available The present article represents an excerpt from the final chapters of the research study titled "The development of structures in mediaeval manuscript bindings - interdependence with conservatory methods". In it, aims, methods of work, archive and library materials used and directions for conservatory methods are presented. Besides, the research study includes also a historcial overview of book bindings, detailed analysis of separate structural elements in Slovenian mediaeval bindings, comprehensive presentation of separate structures, the techniques of binding and materials of the preserved mediaeval bindings in Slovenian public archives and libraries, terminological dictionary of specific professional terms related to binding as a segment of a book, and a catalogue of all analysed bindings, containing a survey of ajI detectable data, sketches,graphite prints and photographs.

  20. Proteomic identification of novel differentiation plasma protein markers in hypobaric hypoxia-induced rat model.

    Directory of Open Access Journals (Sweden)

    Yasmin Ahmad

    Full Text Available BACKGROUND: Hypobaric hypoxia causes complex changes in the expression of genes, including stress related genes and corresponding proteins that are necessary to maintain homeostasis. Whereas most prior studies focused on single proteins, newer methods allowing the simultaneous study of many proteins could lead to a better understanding of complex and dynamic changes that occur during the hypobaric hypoxia. METHODS: In this study we investigated the temporal plasma protein alterations of rat induced by hypobaric hypoxia at a simulated altitude of 7620 m (25,000 ft, 282 mm Hg in a hypobaric chamber. Total plasma proteins collected at different time points (0, 6, 12 and 24 h, separated by two-dimensional electrophoresis (2-DE and identified using matrix assisted laser desorption ionization time of flight (MALDI-TOF/TOF. Biological processes that were enriched in the plasma proteins during hypobaric hypoxia were identified using Gene Ontology (GO analysis. According to their properties and obvious alterations during hypobaric hypoxia, changes of plasma concentrations of Ttr, Prdx-2, Gpx -3, Apo A-I, Hp, Apo-E, Fetub and Nme were selected to be validated by Western blot analysis. RESULTS: Bioinformatics analysis of 25 differentially expressed proteins showed that 23 had corresponding candidates in the database. The expression patterns of the eight selected proteins observed by Western blot were in agreement with 2-DE results, thus confirming the reliability of the proteomic analysis. Most of the proteins identified are related to cellular defense mechanisms involving anti-inflammatory and antioxidant activity. Their presence reflects the consequence of serial cascades initiated by hypobaric hypoxia. CONCLUSION/SIGNIFICANCE: This study provides information about the plasma proteome changes induced in response to hypobaric hypoxia and thus identification of the candidate proteins which can act as novel biomarkers.

  1. plasma treatment

    Directory of Open Access Journals (Sweden)

    Puač Nevena

    2014-11-01

    Full Text Available In this paper we will present results for plasma sterilization of planktonic samples of two reference strains of bacteria, Pseudomonas aeruginosa ATCC 27853 and Enterococcus faecalis ATCC 29212. We have used a plasma needle as a source of non-equilibrium atmospheric plasma in all treatments. This device is already well characterized by OES, derivative probes and mass spectrometry. It was shown that power delivered to the plasma is bellow 2 W and that it produces the main radical oxygen and nitrogen species believed to be responsible for the sterilization process. Here we will only present results obtained by electron paramagnetic resonance which was used to detect the OH, H and NO species. Treatment time and power delivered to the plasma were found to have the strongest influence on sterilization. In all cases we have observed a reduction of several orders of magnitude in the concentration of bacteria and for the longest treatment time complete eradication. A more efficient sterilization was achieved in the case of gram negative bacteria.

  2. Plasma metallization

    CERN Document Server

    Crowther, J M

    1997-01-01

    Many methods are currently used for the production of thin metal films. However, all of these have drawbacks associated with them, for example the need for UHV conditions, high temperatures, exotic metal precursors, or the inability to coat complex shaped objects. Reduction of supported metal salts by non-isothermal plasma treatment does not suffer from these drawbacks. In order to produce and analyse metal films before they become contaminated, a plasma chamber which could be attached directly to a UHV chamber with XPS capability was designed and built. This allowed plasma treatment of supported metal salts and surface analysis by XPS to be performed without exposure of the metal film to the atmosphere. Non-equilibrium plasma treatment of Nylon 66 supported gold(lll) chloride using hydrogen as the feed gas resulted in a 95% pure gold film, the remaining 5% of the film being carbon. If argon or helium were used as the feed gases during plasma treatment the resultant gold films were 100% pure. Some degree of s...

  3. 血浆脑钠肽及脂肪酸结合蛋白水平与慢性收缩性心力衰竭关系的探讨%Investigation of the relationships among plasma B type natriuretic peptide,fatty acid binding protein and chronic congestive heart failure

    Institute of Scientific and Technical Information of China (English)

    曹磊

    2011-01-01

    目的 测定慢性收缩性心力衰竭(CHF)患者的血浆脑钠肽(BNP)及血清脂肪酸结合蛋白(FABP)水平,探讨血浆BNP及血清FABP对CHF诊断及心功能评估的价值.方法 选择住院CHF患者60例,其中NYHA I级12例,II级18例,III级20例,IV级10例;原发疾病:冠心病30例,瓣膜性心脏病9例,扩张型心肌病10例,高血压性心脏病11例.另纳入20例健康体检者为健康对照组,分别测定BNP及FABP浓度,评价心功能.结果 CHF组血浆BNP及血清FABP水平显著高于健康对照组(PNYHA III级>NYHA II级,P均40%患者(P0.05).BNP与FABP呈正相关(r=0.801,P<0.01),两者与LVEF呈负相关(r=-0.718,r=-0.881,P<0.01).结论 血浆BNP及血清FABP测定对CHF的诊断和心功能的分级及预后判断都有重要的临床价值.%Objective To determine the plasma B type natriuretic peptide ( BNP ) and fatty acid binding protein ( FABP ) levels in patients with chronic heart failure ( CHF ), and to investigate the value of plasma BNP and FABP in diagnose of patients with chronic heart failure and in assessment of cardiac function. Methods There were 60 cases of CHF, according to NYHA classification, 12 cases were in degree of Ⅰ, 18 cases were Ⅱ, 20 were Ⅲ, 10 cases in IV. Primary disease:30 cases with CHD,9 cases with valvular heart disease,10 cases with dilated cardiomyopathy, 11 cases with hypertensive heart disease. The plasma BNP and FABP concentration in these 60 patients with chronic heart failure and 20 controls were determined, cardiac function assessment was conducted. Results Level of plasma BNP and FABP in CHF group were significantly higher than that of the control group( P < 0.01 ); and with the increase of degree of heart failure, plasma BNP and FABP levels gradually increased( NYHA Ⅳ > NYHA Ⅲ > NYHA Ⅱ, P < 0.01 ), meanwhile patients with LVEF ≤40% significantly higher than patients with LVEF > 40% ( P < 0.01 ). BNP was positively correlated with FABP( r = 0. 801, P < 0

  4. Ureaplasma urealyticum binds mannose-binding lectin.

    Science.gov (United States)

    Benstein, Barbara D; Ourth, Donald D; Crouse, Dennis T; Shanklin, D Radford

    2004-10-01

    Mannose-binding C-type lectin (MBL) is an important component of innate immunity in mammals. Mannose-binding lectin (MBL), an acute phase protein, acts as an opsonin for phagocytosis and also activates the mannan-binding lectin complement pathway. It may play a particularly significant role during infancy before adequate specific protection can be provided by the adaptive immune system. Ureaplasma urealyticum has been linked to several diseases including pneumonia and chronic lung disease (CLD) in premature infants. We therefore investigated the ability of U. urealyticum to bind MBL. A guinea pig IgG anti-rabbit-MBL antiserum was produced. An immunoblot (dot-blot) assay done on nitrocellulose membrane determined that the anti-MBL antibody had specificity against both rabbit and human MBL. Pure cultures of U. urealyticum, serotype 3, were used to make slide preparations. The slides containing the organisms were then incubated with nonimmune rabbit serum containing MBL. Ureaplasma was shown to bind rabbit MBL with an immunocytochemical assay using the guinea pig IgG anti-rabbit MBL antiserum. Horseradish peroxidase (HRP)-labeled anti-guinea pig IgG was used to localize the reaction. The anti-MBL antiserum was also used in an immunocytochemical assay to localize U. urealyticum in histological sections of lungs from mice specifically infected with this organism. The same method also indicated binding of MBL by ureaplasma in human lung tissue obtained at autopsy from culture positive infants. Our results demonstrate that ureaplasma has the capacity to bind MBL. The absence of MBL may play a role in the predisposition of diseases related to this organism.

  5. Ligand binding mechanics of maltose binding protein.

    Science.gov (United States)

    Bertz, Morten; Rief, Matthias

    2009-11-13

    In the past decade, single-molecule force spectroscopy has provided new insights into the key interactions stabilizing folded proteins. A few recent studies probing the effects of ligand binding on mechanical protein stability have come to quite different conclusions. While some proteins seem to be stabilized considerably by a bound ligand, others appear to be unaffected. Since force acts as a vector in space, it is conceivable that mechanical stabilization by ligand binding is dependent on the direction of force application. In this study, we vary the direction of the force to investigate the effect of ligand binding on the stability of maltose binding protein (MBP). MBP consists of two lobes connected by a hinge region that move from an open to a closed conformation when the ligand maltose binds. Previous mechanical experiments, where load was applied to the N and C termini, have demonstrated that MBP is built up of four building blocks (unfoldons) that sequentially detach from the folded structure. In this study, we design the pulling direction so that force application moves the two MBP lobes apart along the hinge axis. Mechanical unfolding in this geometry proceeds via an intermediate state whose boundaries coincide with previously reported MBP unfoldons. We find that in contrast to N-C-terminal pulling experiments, the mechanical stability of MBP is increased by ligand binding when load is applied to the two lobes and force breaks the protein-ligand interactions directly. Contour length measurements indicate that MBP is forced into an open conformation before unfolding even if ligand is bound. Using mutagenesis experiments, we demonstrate that the mechanical stabilization effect is due to only a few key interactions of the protein with its ligand. This work illustrates how varying the direction of the applied force allows revealing important details about the ligand binding mechanics of a large protein.

  6. Plasma dynamo

    CERN Document Server

    Rincon, F; Schekochihin, A A; Valentini, F

    2015-01-01

    Magnetic fields pervade the entire Universe and, through their dynamical interactions with matter, affect the formation and evolution of astrophysical systems from cosmological to planetary scales. How primordial cosmological seed fields arose and were further amplified to $\\mu$Gauss levels reported in nearby galaxy clusters, near equipartition with kinetic energy of plasma motions and on scales of at least tens of kiloparsecs, is a major theoretical puzzle still largely unconstrained by observations. Extragalactic plasmas are weakly collisional (as opposed to collisional magnetohydrodynamic fluids), and whether magnetic-field growth and its sustainment through an efficient dynamo instability driven by chaotic motions is possible in such plasmas is not known. Fully kinetic numerical simulations of the Vlasov equation in a six-dimensional phase space necessary to answer this question have until recently remained beyond computational capabilities. Here, we show by means of such simulations that magnetic-field a...

  7. 短期膳食赖氨酸摄入量对健康青年男性血浆PA、RBP水平的影响及意义%Effect of different dietary lysine intakes on the plasma prealbumin and retinol-binding protein levels of healthy male youths in short period

    Institute of Scientific and Technical Information of China (English)

    田颖; 彭景; 陈玉; 李哲光; 龚君俊; 丰晔; 陈俊

    2012-01-01

    Objective To provide evidence to establish the lysine reference intakes for different population. Methods Seven healthy young men were selected as subjects. Five dietary lysine intakes of 25 , 35 , 45, 55 , 65 mg/( kg · d) were given to subjects from high to low every week by turns. From the 1st day to the 6th day of each experiment week were adaptation days, and the 7 th day was blood sample collection day. Plasma prealbumin and retinol-binding protein levels were detected by ELISA method. Results The actual dietary lysine intakes were 26, 41, 49, 63, 77 mg/( kg · d). There were no significant differences between the five experimental results and the results before the experiment. Conclusion There is no effect of dietary lysine intakes between 26 to 77 mg/ ( kg · d ) on the plasma prealbumin and retinol-binding protein levels of healthy young male subjects in seven days.%目的 为确定健康人群赖氨酸需要量提供理论依据.方法 选取7例健康青年男性作为研究对象,设定25、35、45、55、65mg/(kg·d)5个膳食赖氨酸水平,由高到低依次给予,每周1个剂量,共5周;每周的第1~6天为适应期,第7天采集受试者静脉血,采用ELISA法检测血浆前白蛋白(PA)、视黄醇结合蛋白(RBP)水平.结果 膳食赖氨酸的实际摄入量为26、41、49、63、77 mg/(kg·d),与实验前相比,摄入5个膳食赖氨酸水平者PA、RBP变化无统计学意义.结论 健康青年男性7天膳食赖氨酸摄入量为26 ~77 mg/(kg·d)不会对血浆PA、RBP水平造成影响.

  8. Study of Plasma Protein Binding Rate and Intestinal Absorption Kinetics of m-Nisoldipine Despinner in Rats%间尼索地平消旋体在大鼠血浆中的蛋白结合率及其肠吸收动力学研究

    Institute of Scientific and Technical Information of China (English)

    李敏; 胡金梅; 王春英; 李小娜; 张志斐; 张兰桐

    2013-01-01

    OBJECTIVE: To study plasma protein binding rate and intestinal absorption kinetics of m-nisoldipine despinner in rats. METHODS: Equilibrium dialysis and HPLC were used to determine the protein binding rates of m-nisoldipine despinner (0.2, 0.5, 1.0, 2.0 μg/ml) in rat plasma. Apparent maximum capacity (βp) of drug binding by albumin and dissociation constant (Kdp) of drug-protein compound were calculated. In intestine reflux test, HPLC method was established for the content determination of m-ni-soldipine despinner and phenolsulfonphthalein in the circulation solution of duodenum, jejunum, ileum and colon to calculate the percentage of drug absorption in 4 h, and the effects of different concentrations of drug, pH value and tween-80 on the absorption kinetics were investigated. RESULTS: The protein, binding rates of m-nisoldipine despinner (0.2, 0.5, 1.0, 2.0 μg/ml) with rat plasma were (96.38 ±3.6)% ,(97.13 ±3.1)%, (97.27 ±2.8)% and(98.22 ± 3.6)%. βp was 4×l0-4 μmol/g and Kdp was 7×l0-4 umol/L. The percentages of drug absorption in 4 h were (51.26± 1.09)%, (47.8516.25)% ,(43.63± 1.986)% and (39.87±5.42)% in duodenum, jejunum, ileum and colon. Intestinal circulation solution with different pH value and different concentrations of tween-80 showed no significant effect on intestinal absorption percentage (P>0.05). A linear relation between the concentration of intestinal circulation solution and absorption percentage was found. CONCLUSIONS: The binding rates of m-nisoldipine despinner with rat plasma protein are in high level in concentration-dependant manner. The absorption of m-nisoldipine in rat's intestine is a first-order process with the passive diffusion mechanism.%目的:研究间尼索地平消旋体在大鼠血浆中的蛋白结合率及其肠吸收动力学.方法:采用平衡透析法和高效液相色谱(HPLC)法测定尼索地平消旋体(0.2、0.5、1.0、2.0 μg/ml)在大鼠血浆中的蛋白结合率,并计算白蛋白结合药物的表

  9. Plasma medicine

    CERN Document Server

    Fridman, Alexander

    2012-01-01

    This comprehensive text is suitable for researchers and graduate students of a 'hot' new topic in medical physics. Written by the world's leading experts,  this book aims to present recent developments in plasma medicine, both technological and scientific, reviewed in a fashion accessible to the highly interdisciplinary audience consisting of doctors, physicists, biologists, chemists and other scientists, university students and professors, engineers and medical practitioners. The book focuses on major topics and covers the physics required to develop novel plasma discharges relevant for medic

  10. The inhibition of anti-DNA binding to DNA by nucleic acid binding polymers.

    Directory of Open Access Journals (Sweden)

    Nancy A Stearns

    Full Text Available Antibodies to DNA (anti-DNA are the serological hallmark of systemic lupus erythematosus (SLE and can mediate disease pathogenesis by the formation of immune complexes. Since blocking immune complex formation can attenuate disease manifestations, the effects of nucleic acid binding polymers (NABPs on anti-DNA binding in vitro were investigated. The compounds tested included polyamidoamine dendrimer, 1,4-diaminobutane core, generation 3.0 (PAMAM-G3, hexadimethrine bromide, and a β-cylodextrin-containing polycation. As shown with plasma from patients with SLE, NABPs can inhibit anti-DNA antibody binding in ELISA assays. The inhibition was specific since the NABPs did not affect binding to tetanus toxoid or the Sm protein, another lupus autoantigen. Furthermore, the polymers could displace antibody from preformed complexes. Together, these results indicate that NABPs can inhibit the formation of immune complexes and may represent a new approach to treatment.

  11. Protein Binding Pocket Dynamics.

    Science.gov (United States)

    Stank, Antonia; Kokh, Daria B; Fuller, Jonathan C; Wade, Rebecca C

    2016-05-17

    The dynamics of protein binding pockets are crucial for their interaction specificity. Structural flexibility allows proteins to adapt to their individual molecular binding partners and facilitates the binding process. This implies the necessity to consider protein internal motion in determining and predicting binding properties and in designing new binders. Although accounting for protein dynamics presents a challenge for computational approaches, it expands the structural and physicochemical space for compound design and thus offers the prospect of improved binding specificity and selectivity. A cavity on the surface or in the interior of a protein that possesses suitable properties for binding a ligand is usually referred to as a binding pocket. The set of amino acid residues around a binding pocket determines its physicochemical characteristics and, together with its shape and location in a protein, defines its functionality. Residues outside the binding site can also have a long-range effect on the properties of the binding pocket. Cavities with similar functionalities are often conserved across protein families. For example, enzyme active sites are usually concave surfaces that present amino acid residues in a suitable configuration for binding low molecular weight compounds. Macromolecular binding pockets, on the other hand, are located on the protein surface and are often shallower. The mobility of proteins allows the opening, closing, and adaptation of binding pockets to regulate binding processes and specific protein functionalities. For example, channels and tunnels can exist permanently or transiently to transport compounds to and from a binding site. The influence of protein flexibility on binding pockets can vary from small changes to an already existent pocket to the formation of a completely new pocket. Here, we review recent developments in computational methods to detect and define binding pockets and to study pocket dynamics. We introduce five

  12. Plasma physics and engineering

    CERN Document Server

    Fridman, Alexander

    2011-01-01

    Part I: Fundamentals of Plasma Physics and Plasma ChemistryPlasma in Nature, in the Laboratory, and in IndustryOccurrence of Plasma: Natural and Man MadeGas DischargesPlasma Applications, Plasmas in IndustryPlasma Applications for Environmental ControlPlasma Applications in Energy ConversionPlasma Application for Material ProcessingBreakthrough Plasma Applications in Modern TechnologyElementary Processes of Charged Species in PlasmaElementary Charged Particles in Plasma and Their Elastic and Inelastic CollisionsIonization ProcessesMechanisms of Electron Losses: The Electron-Ion RecombinationEl

  13. Python bindings for libcloudph++

    OpenAIRE

    Jarecka, Dorota; Arabas, Sylwester; Del Vento, Davide

    2015-01-01

    This technical note introduces the Python bindings for libcloudph++. The libcloudph++ is a C++ library of algorithms for representing atmospheric cloud microphysics in numerical models. The bindings expose the complete functionality of the library to the Python users. The bindings are implemented using the Boost.Python C++ library and use NumPy arrays. This note includes listings with Python scripts exemplifying the use of selected library components. An example solution for using the Python ...

  14. DNS & Bind Cookbook

    CERN Document Server

    Liu, Cricket

    2011-01-01

    The DNS & BIND Cookbook presents solutions to the many problems faced by network administrators responsible for a name server. Following O'Reilly's popular problem-and-solution cookbook format, this title is an indispensable companion to DNS & BIND, 4th Edition, the definitive guide to the critical task of name server administration. The cookbook contains dozens of code recipes showing solutions to everyday problems, ranging from simple questions, like, "How do I get BIND?" to more advanced topics like providing name service for IPv6 addresses. It's full of BIND configuration files that yo

  15. Python bindings for libcloudph++

    CERN Document Server

    Jarecka, Dorota; Del Vento, Davide

    2015-01-01

    This technical note introduces the Python bindings for libcloudph++. The libcloudph++ is a C++ library of algorithms for representing atmospheric cloud microphysics in numerical models. The bindings expose the complete functionality of the library to the Python users. The bindings are implemented using the Boost.Python C++ library and use NumPy arrays. This note includes listings with Python scripts exemplifying the use of selected library components. An example solution for using the Python bindings to access libcloudph++ from Fortran is presented.

  16. Fibronectin binding to protein A-containing staphylococci.

    OpenAIRE

    Doran, J E; Raynor, R H

    1981-01-01

    Fibronectin (Fn) was found to bind to protein A-containing isolates of Staphylococcus aureus, but not to mutant strains devoid of this protein nor to clinical isolates of S. epidermidis. Fn was purified from human plasma by affinity chromatography on gelatin-Sepharose. After elution with 4 M urea, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of purified material detected no immunoglobulin contamination. This purified Fn was radiolabeled with 125I and used in binding assays. Quant...

  17. Magnetoresistive waves in plasmas

    Science.gov (United States)

    Felber, F. S.; Hunter, R. O., Jr.; Pereira, N. R.; Tajima, T.

    1982-10-01

    The self-generated magnetic field of a current diffusing into a plasma between conductors can magnetically insulate the plasma. Propagation of magnetoresistive waves in plasmas is analyzed. Applications to plasma opening switches are discussed.

  18. On Binding Domains

    NARCIS (Netherlands)

    Everaert, M.B.H.

    2005-01-01

    In this paper I want to explore reasons for replacing Binding Theory based on the anaphor-pronoun dichotomy by a Binding Theory allowing more domains restricting/defining anaphoric dependencies. This will, thus, have consequences for the partitioning of anaphoric elements, presupposing more types of

  19. Melanin-binding radiopharmaceuticals

    Energy Technology Data Exchange (ETDEWEB)

    Packer, S; Fairchild, R G; Watts, K P; Greenberg, D; Hannon, S J

    1980-01-01

    The scope of this paper is limited to an analysis of the factors that are important to the relationship of radiopharmaceuticals to melanin. While the authors do not attempt to deal with differences between melanin-binding vs. melanoma-binding, a notable variance is assumed. (PSB)

  20. DNS BIND Server Configuration

    Directory of Open Access Journals (Sweden)

    Radu MARSANU

    2011-01-01

    Full Text Available After a brief presentation of the DNS and BIND standard for Unix platforms, the paper presents an application which has a principal objective, the configuring of the DNS BIND 9 server. The general objectives of the application are presented, follow by the description of the details of designing the program.

  1. Electrosurgical plasmas

    Science.gov (United States)

    Stalder, Kenneth R.; McMillen, Donald F.; Woloszko, Jean

    2005-06-01

    Electrosurgical medical devices based on repetitively pulsed nonequilibrium micron-scale to millimetre-scale plasma discharges in saline solutions are described. The formation of vapour layers (bubbles) around active electrodes appears to be a common feature at moderate (<300 V rms) voltages, and dissociation, excitation and ionization of the vapour in these bubbles produces chemical conditions that are thought to be the source of beneficial tissue removal and treatment. Experimental data are discussed, as are the results of modelling efforts of the plasma chemistry. Hydroxyl radicals, hydrogen atoms and other species are observed spectroscopically and their interactions with collagen, a common component of tissue encountered in surgical situations, are considered. Several pathways by which hydroxyl radicals interacting with collagen can lead to tissue removal are discussed.

  2. Electrosurgical plasmas

    Energy Technology Data Exchange (ETDEWEB)

    Stalder, Kenneth R; McMillen, Donald F; Woloszko, Jean [ArthroCare Corp., Sunnyvale, CA 94085-3523 (United States)

    2005-06-07

    Electrosurgical medical devices based on repetitively pulsed nonequilibrium micron-scale to millimetre-scale plasma discharges in saline solutions are described. The formation of vapour layers (bubbles) around active electrodes appears to be a common feature at moderate (<300 V rms) voltages, and dissociation, excitation and ionization of the vapour in these bubbles produces chemical conditions that are thought to be the source of beneficial tissue removal and treatment. Experimental data are discussed, as are the results of modelling efforts of the plasma chemistry. Hydroxyl radicals, hydrogen atoms and other species are observed spectroscopically and their interactions with collagen, a common component of tissue encountered in surgical situations, are considered. Several pathways by which hydroxyl radicals interacting with collagen can lead to tissue removal are discussed.

  3. Fibrinogen binds to nontoxigenic and toxigenic Corynebacterium diphtheriae strains

    Directory of Open Access Journals (Sweden)

    Priscila Soares Sabbadini

    2010-08-01

    Full Text Available The production of fibrinous exudates may play an important role in determining the outcome of bacterial infection. Although pseudomembrane formation is a characteristic feature of diphtheria, little is known about the fibrinogen (Fbn-binding properties of Corynebacterium diphtheriae strains and the influence of the gene that codes for diphtheria toxin (tox gene in this process. In this study we demonstrated the ability of C. diphtheriae strains to bind to Fbn and to convert Fbn to fibrin. Bacterial interaction with rabbit plasma was evaluated by both slide and tube tests. Interaction of microorganisms with human Fbn was evaluated by both enzyme linked immunosorbent assay (ELISA and fluorescein isothiocyanate-conjugated (FITC Fbn binding assays. Nontoxigenic and toxigenic strains formed bacterial aggregates in the presence of plasma in the slide tests. The ability to convert Fbn to a loose web of fibrin in the plasma solution in the tube tests appeared to be a common characteristic of the species, including strains that do not carry the tox gene. Fbn binding to C. diphtheriae strains occurred at varying intensities, as demonstrated by the FITC-Fbn and ELISA binding assays. Our data suggest that the capacity to bind to Fbn and to convert Fbn to fibrin may play a role in pseudomembrane formation and act as virulence determinants of both nontoxigenic and toxigenic strains.

  4. Fibrinogen binds to nontoxigenic and toxigenic Corynebacterium diphtheriae strains.

    Science.gov (United States)

    Sabbadini, Priscila Soares; Genovez, Marcia Rocha Novais; Silva, Cecília Ferreira da; Adelino, Thelma Lúcia Novaes; Santos, Cintia Silva dos; Pereira, Gabriela Andrade; Nagao, Prescilla Emy; Dias, Alexandre Alves de Souza de Oliveira; Mattos-Guaraldi, Ana Luiza; Hirata Júnior, Raphael

    2010-08-01

    The production of fibrinous exudates may play an important role in determining the outcome of bacterial infection. Although pseudomembrane formation is a characteristic feature of diphtheria, little is known about the fibrinogen (Fbn)-binding properties of Corynebacterium diphtheriae strains and the influence of the gene that codes for diphtheria toxin (tox gene) in this process. In this study we demonstrated the ability of C. diphtheriae strains to bind to Fbn and to convert Fbn to fibrin. Bacterial interaction with rabbit plasma was evaluated by both slide and tube tests. Interaction of microorganisms with human Fbn was evaluated by both enzyme linked immunosorbent assay (ELISA) and fluorescein isothiocyanate-conjugated (FITC) Fbn binding assays. Nontoxigenic and toxigenic strains formed bacterial aggregates in the presence of plasma in the slide tests. The ability to convert Fbn to a loose web of fibrin in the plasma solution in the tube tests appeared to be a common characteristic of the species, including strains that do not carry the tox gene. Fbn binding to C. diphtheriae strains occurred at varying intensities, as demonstrated by the FITC-Fbn and ELISA binding assays. Our data suggest that the capacity to bind to Fbn and to convert Fbn to fibrin may play a role in pseudomembrane formation and act as virulence determinants of both nontoxigenic and toxigenic strains.

  5. Thermodynamics of fragment binding.

    Science.gov (United States)

    Ferenczy, György G; Keserű, György M

    2012-04-23

    The ligand binding pockets of proteins have preponderance of hydrophobic amino acids and are typically within the apolar interior of the protein; nevertheless, they are able to bind low complexity, polar, water-soluble fragments. In order to understand this phenomenon, we analyzed high resolution X-ray data of protein-ligand complexes from the Protein Data Bank and found that fragments bind to proteins with two near optimal geometry H-bonds on average. The linear extent of the fragment binding site was found not to be larger than 10 Å, and the H-bonding region was found to be restricted to about 5 Å on average. The number of conserved H-bonds in proteins cocrystallized with multiple different fragments is also near to 2. These fragment binding sites that are able to form limited number of strong H-bonds in a hydrophobic environment are identified as hot spots. An estimate of the free-energy gain of H-bond formation versus apolar desolvation supports that fragment sized compounds need H-bonds to achieve detectable binding. This suggests that fragment binding is mostly enthalpic that is in line with their observed binding thermodynamics documented in Isothermal Titration Calorimetry (ITC) data sets and gives a thermodynamic rationale for fragment based approaches. The binding of larger compounds tends to more rely on apolar desolvation with a corresponding increase of the entropy content of their binding free-energy. These findings explain the reported size-dependence of maximal available affinity and ligand efficiency both behaving differently in the small molecule region featured by strong H-bond formation and in the larger molecule region featured by apolar desolvation.

  6. Plasma physics

    CERN Document Server

    Cairns, R A

    1985-01-01

    This book is intended as an introduction to plasma physics at a level suitable for advanced undergraduates or beginning postgraduate students in physics, applied mathematics or astrophysics. The main prerequisite is a knowledge of electromagnetism and of the associated mathematics of vector calculus. SI units are used throughout. There is still a tendency amongst some plasma physics researchers to· cling to C.g.S. units, but it is the author's view that universal adoption of SI units, which have been the internationally agreed standard since 1960, is to be encouraged. After a short introductory chapter, the basic properties of a plasma con­ cerning particle orbits, fluid theory, Coulomb collisions and waves are set out in Chapters 2-5, with illustrations drawn from problems in nuclear fusion research and space physics. The emphasis is on the essential physics involved and (he theoretical and mathematical approach has been kept as simple and intuitive as possible. An attempt has been made to draw attention t...

  7. Plasma pharmacy - physical plasma in pharmaceutical applications.

    Science.gov (United States)

    von Woedtke, Th; Haertel, B; Weltmann, K-D; Lindequist, U

    2013-07-01

    During the last years the use of physical plasma for medical applications has grown rapidly. A multitude of findings about plasma-cell and plasma-tissue interactions and its possible use in therapy have been provided. One of the key findings of plasma medical basic research is that several biological effects do not result from direct plasma-cell or plasma-tissue interaction but are mediated by liquids. Above all, it was demonstrated that simple liquids like water or physiological saline, are antimicrobially active after treatment by atmospheric pressure plasma and that these effects are attributable to the generation of different low-molecular reactive species. Besides, it could be shown that plasma treatment leads to the stimulation of specific aspects of cell metabolism and to a transient and reversible increase of diffusion properties of biological barriers. All these results gave rise to think about another new and innovative field of medical plasma application. In contrast to plasma medicine, which means the direct use of plasmas on or in the living organism for direct therapeutic purposes, this field - as a specific field of medical plasma application - is called plasma pharmacy. Based on the present state of knowledge, most promising application fields of plasma pharmacy might be: plasma-based generation of biologically active liquids; plasma-based preparation, optimization, or stabilization of - mainly liquid - pharmaceutical preparations; support of drug transport across biological barriers; plasma-based stimulation of biotechnological processes.

  8. Mannan-binding lectin and healing of a radiation-induced chronic ulcer--a case report on mannan-binding lectin replacement therapy

    DEFF Research Database (Denmark)

    Maaløe, Nanna; Bonde, C; Laursen, I

    2011-01-01

    Mannan-binding lectin is an important component of innate immunity, and insufficiency is associated with several clinical disorders. Recently, experimental replacement therapy with plasma-derived mannan-binding lectin has become an option. The current article presents the case of a patient with a...... that deficiency of mannan-binding lectin can explain cases of otherwise unexplained impaired healing, and that replacement therapy is considered in such cases.......Mannan-binding lectin is an important component of innate immunity, and insufficiency is associated with several clinical disorders. Recently, experimental replacement therapy with plasma-derived mannan-binding lectin has become an option. The current article presents the case of a patient...... with an insufficient level of mannan-binding lectin and a chronic radiation-induced ulcer following the treatment of breast cancer. After 15 months of initially conservative treatment and thereafter plastic surgery, the healing was still impaired with necrosis in the periphery of the ulcer. Immunological work...

  9. Goodpasture Antigen-binding Protein/Ceramide Transporter Binds to Human Serum Amyloid P-Component and Is Present in Brain Amyloid Plaques

    NARCIS (Netherlands)

    Mencarelli, Chiara; Bode, Gerard H.; Losen, Mario; Kulharia, Mahesh; Molenaar, Peter C.; Veerhuis, Robert; Steinbusch, Harry W. M.; De Baets, Marc H.; Nicolaes, Gerry A. F.; Martinez-Martinez, Pilar

    2012-01-01

    Serum amyloid P component (SAP) is a non-fibrillar glycoprotein belonging to the pentraxin family of the innate immune system. SAP is present in plasma, basement membranes, and amyloid deposits. This study demonstrates, for the first time, that the Goodpasture antigen-binding protein (GPBP) binds to

  10. Development of atmospheric pressure large area plasma jet for sterilisation and investigation of molecule and plasma interaction

    Science.gov (United States)

    Zerbe, Kristina; Iberler, Marcus; Jacoby, Joachim; Wagner, Christopher

    2016-09-01

    The intention of the project is the development and improvement of an atmospheric plasma jet based on various discharge forms (e.g. DBD, RF, micro-array) for sterilisation of biomedical equipment and investigation of biomolecules under the influence of plasma stress. The major objective is to design a plasma jet with a large area and an extended length. Due to the success on small scale plasma sterilisation the request of large area plasma has increased. Many applications of chemical disinfection in environmental and medical cleaning could thereby be complemented. Subsequently, the interaction between plasma and biomolecules should be investigated to improve plasma strerilisation. Special interest will be on non equilibrium plasma electrons affecting the chemical bindings of organic molecules.

  11. Mechanism of Action and Clinical Application of Tafamidis in Hereditary Transthyretin Amyloidosis

    OpenAIRE

    Coelho, Teresa; Merlini, Giampaolo; Bulawa, Christine E.; Fleming, James A.; Judge, Daniel P.; Kelly, Jeffery W.; Maurer, Mathew S.; Planté-Bordeneuve, Violaine; Labaudinière, Richard; Mundayat, Rajiv; Riley, Steve; Lombardo, Ilise; Huertas, Pedro

    2016-01-01

    Transthyretin (TTR) transports the retinol-binding protein–vitamin A complex and is a minor transporter of thyroxine in blood. Its tetrameric structure undergoes rate-limiting dissociation and monomer misfolding, enabling TTR to aggregate or to become amyloidogenic. Mutations in the TTR gene generally destabilize the tetramer and/or accelerate tetramer dissociation, promoting amyloidogenesis. TTR-related amyloidoses are rare, fatal, protein-misfolding disorders, characterized by formation of ...

  12. Atovaquone and quinine anti-malarials inhibit ATP binding cassette transporter activity

    NARCIS (Netherlands)

    Rijpma, S.R.; Heuvel, J.J.; Velden, M. van der; Sauerwein, R.W.; Russel, F.G.; Koenderink, J.B.

    2014-01-01

    BACKGROUND: Therapeutic blood plasma concentrations of anti-malarial drugs are essential for successful treatment. Pharmacokinetics of pharmaceutical compounds are dependent of adsorption, distribution, metabolism, and excretion. ATP binding cassette (ABC) transport proteins are particularly involve

  13. Serum mannan-binding lectin-associated serine protease 2 levels in colorectal cancer: relation to recurrence and mortality

    DEFF Research Database (Denmark)

    Ytting, Henriette; Christensen, Ib Jarle; Thiel, Steffen;

    2005-01-01

    PURPOSE: Mannan-binding lectin-associated serine protease 2 (MASP-2) is a plasma protein involved in inflammatory processes. MASP-2 circulates in complex with the protein mannan-binding lectin (MBL) or ficolins, and is activated to recruit the complement system when MBL binds to its targets...

  14. Angiotensin II Inhibits Insulin Binding to Endothelial Cells

    Directory of Open Access Journals (Sweden)

    Su-Jin Oh

    2011-06-01

    Full Text Available BackgroundInsulin-mediated glucose uptake in insulin target tissues is correlated with interstitial insulin concentration, rather than plasma insulin concentration. Therefore, insulin delivery to the interstitium of target tissues is very important, and the endothelium may also play an important role in the development of insulin resistance.MethodsAfter treating bovine aortic endothelial cells with angiotensin II (ATII, we observed the changes in insulin binding capacity and the amounts of insulin receptor (IR on the cell membranes and in the cytosol.ResultsAfter treatment of 10-7M ATII, insulin binding was decreased progressively, up to 60% at 60 minutes (P<0.05. ATII receptor blocker (eprosartan dose dependently improved the insulin binding capacity which was reduced by ATII (P<0.05. At 200 µM, eprosartan fully restored insulin binding capacity, althogh it resulted in only a 20% to 30% restoration at the therapeutic concentration. ATII did not affect the total amount of IR, but it did reduce the amount of IR on the plasma membrane and increased that in the cytosol.ConclusionATII decreased the insulin binding capacity of the tested cells. ATII did not affect the total amount of IR but did decrease the amount of IR on the plasma membrane. Our data indicate that ATII decreases insulin binding by translocating IR from the plasma membrane to the cytosol. The binding of insulin to IR is important for insulin-induced vasodilation and transendothelial insulin transport. Therefore, ATII may cause insulin resistance through this endothelium-based mechanism.

  15. Binding of human serum albumin to PEGylated liposomes: insights into binding numbers and dynamics by fluorescence correlation spectroscopy

    DEFF Research Database (Denmark)

    Kristensen, Kasper; Urquhart, Andrew; Thormann, Esben

    2016-01-01

    understood. For example, there is generally a lack of knowledge about the liposome binding affinities and dynamics of common types of blood plasma proteins. Fluorescence correlation spectroscopy (FCS) is a powerful experimental technique that potentially can provide such knowledge. In this study, we have...... used FCS to investigate the binding of human serum albumin (HSA) to standard types of PEGylated fluid-phase liposomes (consisting of DOPC and DOPE-PEG2k) and PEGylated gel-phase liposomes (consisting of DSPC and DSPE-PEG2k) with various PEG chain surface densities. We detected no significant binding...

  16. SHBG (Sex Hormone Binding Globulin)

    Science.gov (United States)

    ... as: Testosterone-estrogen Binding Globulin; TeBG Formal name: Sex Hormone Binding Globulin Related tests: Testosterone , Free Testosterone, ... I should know? How is it used? The sex hormone binding globulin (SHBG) test may be used ...

  17. Analysis of clinical characteristics and plasma level of mannose-binding lectin in 68 children with febrile convulsion%68例热性惊厥患儿临床特点及其血浆甘露糖结合凝集素水平变化

    Institute of Scientific and Technical Information of China (English)

    王超前; 华春珍; 罗巧二; 王琦婧; 戴凯丽; 黄邢邢; 杨钦梳

    2012-01-01

    [Objective] To analyze the clinical characteristics of febrile convulsion (FC) occurred in children and study the mannose-binding lectin (MBL) levels in plasma at both acute and convalescent stages in the case group. [Methods] Plasma samples were separated from whole blood which was collected from children with FC at the acute and convalescent stages during August 2010 to July 2011. The MBL concentrations were measured by ELISA method and the SPSS 11. 0 software was used in the study. [Results] Among all 68 cases,The mean MBL levels were (595 + 559)ng/mL at acute stage and (585 ± 466)ng/mL in recovery stage,and no significant difference was found between the two groups (Z = 0. 207,P = 0. 836). However,there was a significant correlation of the MBL levels between the two stages (r=0. 809, P<0. 01). 29. 4% of all individuals had low MBL levels less than 200 ng/mL at convalescent stage. Acute reaction of MBL was observed in 38. 2% children with FC. No correlation was found between MBL levels and CRP levels(r =-0. 15,P=0. 22). It was found that 89. 7% of the FC children was caused by acute upper respiratory tract infection. The creatine kinase-MB exceeding normal value was found in 54. 1% of all individuals. [Conclusion] As a whole, convulsion acting as a kind of stress, had no significant effects on the MBL level in vivo.%[目的]了解热性惊厥患儿的临床特点及其血浆甘露糖结合凝集素(mannose-binding lectin,MBL)水平在惊厥前后的变化. [方法]对2010年8月-2011年7月在本院住院的68例热性惊厥患儿,应用ELISA法检测急性期与恢复期血浆MBL浓度,同时进行临床特点分析.统计分析采用SPSS 11.0. [结果]急性期和恢复期MBL平均浓度分别为(595±559) ng/mL和(585±466) ng/mL,二者差异无统计学意义(Z=0.207,P=0.836),急性期与恢复期MBL水平呈线性相关(r=0.809,P<0.01).其中恢复期MBL血浆浓度<200 ng/mL的病例占29.4%,MBL参与急性应答占全部病例的38.2%.MBL浓

  18. Microbes bind complement inhibitor factor H via a common site.

    Directory of Open Access Journals (Sweden)

    T Meri

    Full Text Available To cause infections microbes need to evade host defense systems, one of these being the evolutionarily old and important arm of innate immunity, the alternative pathway of complement. It can attack all kinds of targets and is tightly controlled in plasma and on host cells by plasma complement regulator factor H (FH. FH binds simultaneously to host cell surface structures such as heparin or glycosaminoglycans via domain 20 and to the main complement opsonin C3b via domain 19. Many pathogenic microbes protect themselves from complement by recruiting host FH. We analyzed how and why different microbes bind FH via domains 19-20 (FH19-20. We used a selection of FH19-20 point mutants to reveal the binding sites of several microbial proteins and whole microbes (Haemophilus influenzae, Bordetella pertussis, Pseudomonas aeruginosa, Streptococcus pneumonia, Candida albicans, Borrelia burgdorferi, and Borrelia hermsii. We show that all studied microbes use the same binding region located on one side of domain 20. Binding of FH to the microbial proteins was inhibited with heparin showing that the common microbial binding site overlaps with the heparin site needed for efficient binding of FH to host cells. Surprisingly, the microbial proteins enhanced binding of FH19-20 to C3b and down-regulation of complement activation. We show that this is caused by formation of a tripartite complex between the microbial protein, FH, and C3b. In this study we reveal that seven microbes representing different phyla utilize a common binding site on the domain 20 of FH for complement evasion. Binding via this site not only mimics the glycosaminoglycans of the host cells, but also enhances function of FH on the microbial surfaces via the novel mechanism of tripartite complex formation. This is a unique example of convergent evolution resulting in enhanced immune evasion of important pathogens via utilization of a "superevasion site."

  19. 液质联用超滤法测定注射用复方荭草中原儿茶酸、异荭草素及野黄芩苷的血浆蛋白结合率%Determination of Plasma Protein Binding Rates of Protocatechuic Acid, Isoorientin and Scutellarin in Compound Hongcao Freeze-Dried Powder for Injection by Ultrafiltration Combined with UPLC-ESI-MS/MS

    Institute of Scientific and Technical Information of China (English)

    黄勇; 陈慧; 郑林; 何峰; 张治蓉; 王永林

    2011-01-01

    OBJECTIVE To determine the plasma protein binding rates of protocatechuic acid, isoorientin and scutellarin in Compound Hongcao freeze-dried powder for injection by ultrafiltration method for providing on the safe use of medication. METHODS Ultrafiltration combined with liquid chromatography-tandem mass spectrometry (UPLC-ESI-MS/MS) was employed to determine the protein binding rats of protocatechuic acid, isoorientin and scutellarin in the compound hongcao freeze-dried powder. When determining the concentrations of the 3 compounds, 2 kinds of matrix as plasma and buffer solution were chosen. RESULTS The average human protein binding rates of protocatechuic acid, isoorientin and Scutellarin were (65. 33 ± 0.61) % , (85. 97 ± 0. 43)% and(90. 09 ± 0. 28 ) % with the drugplasma concentrations of 5,10,25,50 and 100 μg · mL-1, respectively. CONCLUSION The current study shows the medium binding ability of protocatechuic acid and high binding ability of isoorientin and scutellarin to plasma protein. The method has high sensitivity, good reproduction, with simple management thus fulfilling the requirement The plsma protein bindings of the three compounds were independent of the investigated concentrations.%目的 测定注射用复方荭草中原儿茶酸、异荭草素及野黄芩苷的血浆蛋白结合率,为临床安全用药提供参考.方法 以超高效液相色谱-质谱联用为检测手段,结合超滤离心法,由于基质背景的差异分别建立血浆样本及超滤液中的含量测定方法考察原儿茶酸、异荭草素及野黄芩苷与血浆蛋白的结合情况.结果 注射用复方荭草在人血浆中浓度为5~100 μg· mL-1时,原儿茶酸、异荭草素及野黄芩苷与血浆蛋白的平均结合率分别为(65.33±0.61)%+(85.97±0.43)%,(90.09±0.28)%.结论 原儿茶酸与血浆具有中等强度的结合,异荭草素及野黄芩苷与血浆结合力较强,它们与血浆蛋白结合能力在考察的浓度范围内无浓度

  20. Insulin-induced lipid binding to hemoglobin

    Directory of Open Access Journals (Sweden)

    VESNA NIKETIC

    2003-01-01

    Full Text Available Under hypoglycemic conditions, concomitant hyperinsulinism causes an apparent modification of hemoglobin (Hb which is manifested by its aggregation (Niketi} et al., Clin. Chim. Acta 197 (1991 47. In the present work the causes and mechanisms underlying this Hb modification were studied. Hemoglobin isolated from normal erythrocytes incubated with insulin was analyzed by applying 31P-spectrometry and lipid extraction and analysis. To study the dynamics of the plasma membrane during hyperinsulinism, a fluorescent lipid-analog was applied. In the presence of insulin, phosphatidylserine (PS, phosphatidylethanolamine (PE and cholesterol were found to bind to Hb. Lipid binding resulted in Hb aggregation, a condition that can be reproduced when phospholipids are incubated with Hb in vitro. Using a fluorescent lipid-analog, it was also shown that exposing erythrocytes to supraphysiological concentrations of insulin in vitro resulted in the internalization of lipids. The results presented in this work may have relevance to cases of diabetes mellitus and hypoglycemia.

  1. CARBOHYDRATE-CONTAINING COMPOUNDS WHICH BIND TO CARBOHYDRATE BINDING RECEPTORS

    DEFF Research Database (Denmark)

    1995-01-01

    Carbohydrate-containing compounds which contain saccharides or derivatives thereof and which bind to carbohydrate binding receptors are useful in pharmaceutical products for treatment of inflammatory diseases and other diseases.......Carbohydrate-containing compounds which contain saccharides or derivatives thereof and which bind to carbohydrate binding receptors are useful in pharmaceutical products for treatment of inflammatory diseases and other diseases....

  2. Tafamidis, a potent and selective transthyretin kinetic stabilizer that inhibits the amyloid cascade.

    Science.gov (United States)

    Bulawa, Christine E; Connelly, Stephen; Devit, Michael; Wang, Lan; Weigel, Charlotte; Fleming, James A; Packman, Jeff; Powers, Evan T; Wiseman, R Luke; Foss, Theodore R; Wilson, Ian A; Kelly, Jeffery W; Labaudinière, Richard

    2012-06-12

    The transthyretin amyloidoses (ATTR) are invariably fatal diseases characterized by progressive neuropathy and/or cardiomyopathy. ATTR are caused by aggregation of transthyretin (TTR), a natively tetrameric protein involved in the transport of thyroxine and the vitamin A-retinol-binding protein complex. Mutations within TTR that cause autosomal dominant forms of disease facilitate tetramer dissociation, monomer misfolding, and aggregation, although wild-type TTR can also form amyloid fibrils in elderly patients. Because tetramer dissociation is the rate-limiting step in TTR amyloidogenesis, targeted therapies have focused on small molecules that kinetically stabilize the tetramer, inhibiting TTR amyloid fibril formation. One such compound, tafamidis meglumine (Fx-1006A), has recently completed Phase II/III trials for the treatment of Transthyretin Type Familial Amyloid Polyneuropathy (TTR-FAP) and demonstrated a slowing of disease progression in patients heterozygous for the V30M TTR mutation. Herein we describe the molecular and structural basis of TTR tetramer stabilization by tafamidis. Tafamidis binds selectively and with negative cooperativity (K(d)s ~2 nM and ~200 nM) to the two normally unoccupied thyroxine-binding sites of the tetramer, and kinetically stabilizes TTR. Patient-derived amyloidogenic variants of TTR, including kinetically and thermodynamically less stable mutants, are also stabilized by tafamidis binding. The crystal structure of tafamidis-bound TTR suggests that binding stabilizes the weaker dimer-dimer interface against dissociation, the rate-limiting step of amyloidogenesis.

  3. Tetranectin Binds to the Kringle 1-4 Form of Angiostatin and Modifies Its Functional Activity

    DEFF Research Database (Denmark)

    Mogues, Tirsit; Etzerodt, Michael; Hall, Crystal

    2004-01-01

    Tetranectin is a plasminogen kringle 4 domain-binding protein present in plasma and various tissue locations. Decreased plasma tetranectin or increased tetranectin in stroma of cancers correlates with cancer progression and adverse prognosis. A possible mechanism through which tetranectin could i...

  4. Interspecies In Vitro Evaluation of Stereoselective Protein Binding for 3,4-Methylenedioxymethamphetamine

    Directory of Open Access Journals (Sweden)

    Wan Raihana Wan Aasim

    2017-01-01

    Full Text Available Abuse of 3,4-methylenedioxymethamphetamine (MDMA is becoming more common worldwide. To date, there is no information available on stereoselectivity of MDMA protein binding in humans, rats, and mice. Since stereoselectivity plays an important role in MDMA’s pharmacokinetics and pharmacodynamics, in this study we investigated its stereoselectivity in protein binding. The stereoselective protein binding of rac-MDMA was investigated using two different concentrations (20 and 200 ng/mL in human plasma and mouse and rat sera using an ultrafiltration technique. No significant stereoselectivity in protein binding was observed in both human plasma and rat serum; however, a significant stereoselective binding (p<0.05 was observed in mouse serum. Since the protein binding of MDMA in mouse serum is considerably lower than in humans and rats, caution should be exercised when using mice for in vitro studies involving MDMA.

  5. Acyl-CoA binding proteins; structural and functional conservation over 2000 MYA

    DEFF Research Database (Denmark)

    Faergeman, Nils J; Wadum, Majken; Feddersen, Søren

    2007-01-01

    -CoA binding protein, ACBP, has been proposed to play a pivotal role in the intracellular trafficking and utilization of long-chain fatty acyl-CoA esters. Depletion of acyl-CoA binding protein in yeast results in aberrant organelle morphology incl. fragmented vacuoles, multi-layered plasma membranes...

  6. 21 CFR 862.1685 - Thyroxine-binding globulin test system.

    Science.gov (United States)

    2010-04-01

    ... protein which binds thyroxine, in serum and plasma. Measurements obtained by this device are used in the... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Thyroxine-binding globulin test system. 862.1685 Section 862.1685 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN...

  7. Plasma Free Metanephrines

    Science.gov (United States)

    ... be limited. Home Visit Global Sites Search Help? Plasma Free Metanephrines Share this page: Was this page helpful? Also known as: Plasma Metanephrines Formal name: Fractionated Plasma Free Metanephrines (Metanephrine ...

  8. Improved plasma accelerator

    Science.gov (United States)

    Cheng, D. Y.

    1971-01-01

    Converging, coaxial accelerator electrode configuration operates in vacuum as plasma gun. Plasma forms by periodic injections of high pressure gas that is ionized by electrical discharges. Deflagration mode of discharge provides acceleration, and converging contours of plasma gun provide focusing.

  9. Cortisol levels, binding, and properties of corticosteroid-binding globulin in the serum of primates.

    Science.gov (United States)

    Klosterman, L L; Murai, J T; Siiteri, P K

    1986-01-01

    New World primates have exceptionally high plasma levels of cortisol and other steroid hormones when compared with humans and other primates. It has been suggested that this difference can be explained by either low affinity or concentration of cellular steroid receptors. We have assessed cortisol availability in serum from several species of New and Old World primates under physiological conditions (whole serum at 37 degrees C). Measurements were made of total and free cortisol, corticosteroid-binding globulin (CBG) binding capacity and affinity for cortisol, distribution of cortisol in serum, and its binding to albumin. In agreement with earlier reports, plasma free cortisol levels in Old World primates, prosimians, and humans range from 10-300 nM. However, very high total plasma cortisol together with low CBG binding capacity and affinity result in free cortisol concentrations of 1-4 microM in some New World primates (squirrel monkey and marmosets) but not in others such as the titi and capuchin. In squirrel monkeys, free cortisol levels are far greater than might be predicted from the affinity of the glucocorticoid receptor estimated in cultured skin fibroblasts. In addition to low affinity, CBG from squirrel monkeys and other New World primates exhibits differences in electrophoretic mobility and sedimentation behavior in sucrose density ultracentrifugation, suggestive of a molecular weight that is approximately twice that of CBG from other species. Together with other data these results indicate that the apparent glucocorticoid resistance found in New World primates is a complex phenomenon that is not easily explained by present concepts of glucocorticoid action.

  10. Plasma physics and fusion plasma electrodynamics

    CERN Document Server

    Bers, Abraham

    2016-01-01

    Plasma is a ubiquitous state of matter at high temperatures. The electrodynamics of plasmas encompasses a large number of applications, from understanding plasmas in space and the stars, to their use in processing semiconductors, and their role in controlled energy generation by nuclear fusion. This book covers collective and single particle dynamics of plasmas for fully ionized as well as partially ionized plasmas. Many aspects of plasma physics in current fusion energy generation research are addressed both in magnetic and inertial confinement plasmas. Linear and nonlinear dynamics in hydrodynamic and kinetic descriptions are offered, making both simple and complex aspects of the subject available in nearly every chapter. The approach of dividing the basic aspects of plasma physics as "linear, hydrodynamic descriptions" to be covered first because they are "easier", and postponing the "nonlinear and kinetic descriptions" for later because they are "difficult" is abandoned in this book. For teaching purpose...

  11. Chromatofocusing in the purification and separation of apo- and holo-(vitamin D-binding protein).

    OpenAIRE

    Keenan, M J; Holmes, R. P.

    1985-01-01

    Chromatofocusing was used to purify the vitamin D-binding protein (DBP) from pig plasma in a procedure that consisted of an initial DEAE-cellulose chromatography followed by DEAE-Sephadex chromatography, with final purification by chromatofocusing. The protein was purified 184-fold over its concentration in plasma. When the plasma was labelled with a tracer concentration of [3H]calcidiol, it was apparent that holo- and apo-DBP did not co-chromatograph on chromatofocusing. The separation of th...

  12. Signal transduction of erythrocytes after specific binding of ecdysterone and cholesterol immobilized on nanodispersed magnetite

    Energy Technology Data Exchange (ETDEWEB)

    Mykhaylyk, O.M. E-mail: helek@iptelecom.net.ua; Kotzuruba, A.V.; Buchanevich, O.M.; Korduban, A.M.; Meged, E.F.; Gulaya, N.M

    2001-07-01

    Concurrent binding of cholesterol and ecdysterone immobilized on nanodispersed magnetite to intact rat erythrocytes was investigated. Several binding components on erythrocyte plasma membrane with different affinities were revealed in the range of 10{sup -15}-10{sup -8} M. The specific binding modulates signal transduction through adenylate cyclase and guanylate cyclase systems as manifested by the decrease in cAMP and increase in cGMP second messenger production.

  13. The effect of polyamines on the binding of anti-DNA antibodies from patients with SLE and normal human subjects.

    Science.gov (United States)

    Wang, Xiao; Stearns, Nancy A; Li, Xingfu; Pisetsky, David S

    2014-07-01

    Antibodies to DNA (anti-DNA) are the serological hallmark of systemic lupus erythematosus (SLE). To elucidate specificity further, the effect of polyamines on the binding of anti-DNA antibodies from patients with lupus was tested by ELISA to calf thymus (CT) DNA; we also assessed the binding of plasmas of patients and normal human subjects (NHS) to Micrococcus luteus (MC) DNA. As these studies showed, spermine can dose-dependently inhibit SLE anti-DNA binding to CT DNA and can promote dissociation of preformed immune complexes. With MC DNA as antigen, spermine failed to inhibit the NHS anti-DNA binding. Studies using plasmas adsorbed to a CT DNA cellulose affinity indicated that SLE plasmas are mixtures of anti-DNA that differ in inhibition by spermine and binding to conserved and non-conserved determinants. Together, these studies demonstrate that spermine can influence the binding of anti-DNA autoantibodies and may contribute to the antigenicity of DNA.

  14. Fusion plasma physics

    CERN Document Server

    Stacey, Weston M

    2012-01-01

    This revised and enlarged second edition of the popular textbook and reference contains comprehensive treatments of both the established foundations of magnetic fusion plasma physics and of the newly developing areas of active research. It concludes with a look ahead to fusion power reactors of the future. The well-established topics of fusion plasma physics -- basic plasma phenomena, Coulomb scattering, drifts of charged particles in magnetic and electric fields, plasma confinement by magnetic fields, kinetic and fluid collective plasma theories, plasma equilibria and flux surface geometry, plasma waves and instabilities, classical and neoclassical transport, plasma-materials interactions, radiation, etc. -- are fully developed from first principles through to the computational models employed in modern plasma physics. The new and emerging topics of fusion plasma physics research -- fluctuation-driven plasma transport and gyrokinetic/gyrofluid computational methodology, the physics of the divertor, neutral ...

  15. Terms of Binding

    NARCIS (Netherlands)

    Sevcenco, A.

    2006-01-01

    The present dissertation aimed at achieving two goals. First, it constitutes an attempt to widen the search for phenomena that bear relevance to the idea that binding has a syntactic residue and is not, therefore, an exclusively semantic matter. Second, it tried to provide the technical means to acc

  16. Cellulose binding domain proteins

    Science.gov (United States)

    Shoseyov, Oded; Shpiegl, Itai; Goldstein, Marc; Doi, Roy

    1998-01-01

    A cellulose binding domain (CBD) having a high affinity for crystalline cellulose and chitin is disclosed, along with methods for the molecular cloning and recombinant production thereof. Fusion products comprising the CBD and a second protein are likewise described. A wide range of applications are contemplated for both the CBD and the fusion products, including drug delivery, affinity separations, and diagnostic techniques.

  17. Binding and Bulgarian

    NARCIS (Netherlands)

    Schürcks-Grozeva, Lilia Lubomirova

    2003-01-01

    In haar proefschrift analyseert Lilia Schürcks de anaforische verschijnselen in de Bulgaarse taal. Het gaat dan om wederkerende aspecten, uitgedrukt bij woorden als ‘zich’ en ‘elkaar’. De situatie in het Bulgaars blijkt moeilijk in te passen in de klassieke Binding Theory van Noam Chomsky. Bron: RUG

  18. Sequential memory: Binding dynamics

    Science.gov (United States)

    Afraimovich, Valentin; Gong, Xue; Rabinovich, Mikhail

    2015-10-01

    Temporal order memories are critical for everyday animal and human functioning. Experiments and our own experience show that the binding or association of various features of an event together and the maintaining of multimodality events in sequential order are the key components of any sequential memories—episodic, semantic, working, etc. We study a robustness of binding sequential dynamics based on our previously introduced model in the form of generalized Lotka-Volterra equations. In the phase space of the model, there exists a multi-dimensional binding heteroclinic network consisting of saddle equilibrium points and heteroclinic trajectories joining them. We prove here the robustness of the binding sequential dynamics, i.e., the feasibility phenomenon for coupled heteroclinic networks: for each collection of successive heteroclinic trajectories inside the unified networks, there is an open set of initial points such that the trajectory going through each of them follows the prescribed collection staying in a small neighborhood of it. We show also that the symbolic complexity function of the system restricted to this neighborhood is a polynomial of degree L - 1, where L is the number of modalities.

  19. International movement of plasma and plasma contracting.

    Science.gov (United States)

    Farrugia, A

    2005-01-01

    Plasma fractionation is a global business characterised by technological stability, increasing consolidation and a high level of regulatory oversight. All these factors affect the ease with which plasma derivatives can be accessed in the world market. As domestic regulatory measures in the first world blood economies become increasingly resonant to the precautionary approach, the availability of plasma as a raw material, as well as its cost, become an increasingly significant component in the cost of the final product. This decreases the amount of plasma which fractionators are able to allocate for export activities. Also, regulatory standards in the country of manufacture will reflect priorities in that country which may not be similar to those in export markets, but which will affect entry to those markets. While many countries possess a fractionation capacity, the limiting factor in supply worldwide is the amount of plasma available, and nationalistic drivers for each country to have its own plant are inimical to product safety and supply. Rather, the provision of sufficient supplies of domestic plasma should be the focus of resource allocation, with a choice of an appropriate contract fractionator. However, contract fractionation too may be affected by domestic considerations unrelated to the needs of the country of plasma origin. This chapter will review the global plasma market and the influences on plasma and plasma product movement across national borders. Problems in ensuring adequate safety and supply will be identified, and some tentative approaches to the amelioration of current barriers to the provision of plasma derivatives will be outlined.

  20. Decreased protein binding of moxifloxacin in patients with sepsis?

    Directory of Open Access Journals (Sweden)

    Dorn, Christoph

    2017-02-01

    Full Text Available The mean (SD unbound fraction of moxifloxacin in plasma from patients with severe sepsis or septic shock was determined by ultrafiltration to 85.5±3.0% (range 81.9 and 91.6% indicating a decreased protein binding of moxifloxacin in this population compared with the value of 58–60% provided in the Summary of Product Characteristics. However, previous investigations neglected the influence of pH and temperature on the protein binding of moxifloxacin. Maintaining physiological conditions (pH 7.4, 37°C – as in the present study – the unbound fraction of moxifloxacin in plasma from healthy volunteers was 84%. In contrast, the unbound fraction of moxifloxacin was 77% at 4°C and 66–68% in unbuffered plasma or at pH 8.5 in fair agreement with previously published data. PK/PD parameters e.g. AUC/MIC or ratios between interstitial fluid and free plasma concentrations, which were obtained assuming a protein binding rate of moxifloxacin of 40% or more, should be revised.

  1. Communication through Plasma Sheaths

    CERN Document Server

    Korotkevich, A O; Zakharov, V E

    2007-01-01

    We wish to transmit messages to and from a hypersonic vehicle around which a plasma sheath has formed. For long distance transmission, the signal carrying these messages must be necessarily low frequency, typically 2 GHz, to which the plasma sheath is opaque. The idea is to use the plasma properties to make the plasma sheath appear transparent.

  2. Introduction to plasma dynamics

    CERN Document Server

    Morozov, A I

    2013-01-01

    As the twenty-first century progresses, plasma technology will play an increasing role in our lives, providing new sources of energy, ion-plasma processing of materials, wave electromagnetic radiation sources, space plasma thrusters, and more. Studies of the plasma state of matter not only accelerate technological developments but also improve the understanding of natural phenomena. Beginning with an introduction to the characteristics and types of plasmas, Introduction to Plasma Dynamics covers the basic models of classical diffuse plasmas used to describe such phenomena as linear and shock w

  3. Collisionless plasmas in astrophysics

    CERN Document Server

    Belmont, Gerard; Mottez, Fabrice; Pantellini, Filippo; Pelletier, Guy

    2013-01-01

    Collisionless Plasmas in Astrophysics examines the unique properties of media without collisions in plasma physics. Experts in this field, the authors present the first book to concentrate on collisionless conditions in plasmas, whether close or not to thermal equilibrium. Filling a void in scientific literature, Collisionless Plasmas in Astrophysics explains the possibilities of modeling such plasmas, using a fluid or a kinetic framework. It also addresses common misconceptions that even professionals may possess, on phenomena such as "collisionless (Landau) damping". Abundant illustrations

  4. Association between codon 54 polymorphism of intestinal fatty acid-binding protein 2 gene and plasma lipids in middle-aged and old populations%中老年人群小肠脂肪酸结合蛋白FABP2基因54位密码子多态性与血脂水平的关系

    Institute of Scientific and Technical Information of China (English)

    常晓彤; 王振辉; 侯丽娟; 李彪英; 董明纲; 李桂喜

    2008-01-01

    背景:有研究表明,在不同人群中,突变型54T FABP2与血脂障碍以及代谢综合征的其他特征相关.目的: 调查中老年人群小肠脂肪酸结合蛋白FABP2基因多态性频率分布,分析突变型54T FABP2基因与血脂水平的关系.设计:病例-对照分析.单位: 河北北方学院生物化学教研室和解放军第二五一医院检验科.对象:选择2003-10/2005-04在解放军第二五一医院体检中心进行体检的中老年人469名,男217名,平均年龄(56±10)岁;女252名,平均年龄(55±13)岁.除外肝、肾功能异常者,相互间无血缘关系;患者均对本实验均知情同意.实验已经医院伦理委员会批准.方法:①空腹12 h后,采用全自动生化仪(Olympus AU 6400)测定血浆总胆固醇、三酰甘油、高密度脂蛋白胆固醇、低密度脂蛋白胆固醇、载脂蛋白AⅠ、载脂蛋白B水平.②采集空腹静脉血1mL,枸橼酸钠抗凝,分离白细胞,蛋白酶K消化,酚/氯仿抽提基因组DNA.采用PCR-限制性片段长度多态性技术检测各组基因型分布频率.主要观察指标:①血脂水平.②FABP2 54位点基因型分布频率.结果:①频率分布:男性54A/T FABP2基因型频率分布为A/A0.48,A/T 0.42,T/T0.10;女性为A/A0.44,A/T0.46,T/T 0.10.男性和女性突变型54T等位基因频率分别为0.31,0.33.男女间频率分布情况比较,差异无显著性意义(P > 0.05).②血脂:男性,54T等位基因携带者血浆低密度脂蛋白胆固醇、载脂蛋白B高于54A等位基因携带者,差异有显著性意义(P < 0.05);女性,54T等位基因携带者血浆总胆固醇、低密度脂蛋白胆固醇高于54A等位基因携带者,差异有显著性意义(P < 0.05).结论:在中老年人群中,FABP2基因多态性频率分布与性别无关;54T FABP2基因携带者有高血脂特征.%BACKGROUND: Studies have shown that alanine (A) to threonine (T) substitution at codon 54 of intestinal fatty acid-binding protein (FABP2) in different populations

  5. The plasma carboxypeptidases and the regulation of the plasminogen system.

    Science.gov (United States)

    Plow, E F; Allampallam, K; Redlitz, A

    1997-04-01

    Central to the regulation of the plasminogen system, a proteolytic network that mediates degradation of fibrin and facilitates cell migration, is the binding of plasminogen to carboxy-terminal lysines. These residues occur either naturally on plasmin substrates or cell surfaces or are generated as a consequence of partial plasmin degradation. The basic carboxypeptidases of plasma are capable of removing such carboxy-terminal lysines. Carboxypeptidase N, which is constitutively active, suppresses plasminogen binding to cell surfaces; plasma carboxypeptidase B, which must be proteolytically activated, not only suppresses cellular binding of plasminogen but also dampens fibrinolysis. Thus, the plasma carboxypeptidases may constitute an important regulatory pathway for controlling the activity of the plasminogen system in physiologic, pathophysiologic, and pharmacologic circumstances. (Trends Cardiovasc Med 1997;7:71-75). © 1997, Elsevier Science Inc.

  6. Effect of cysteamine on cytosolic somatostatin binding sites in rabbit duodenal mucosa.

    Science.gov (United States)

    Gonzalez-Guijarro, L; Lopez-Ruiz, M P; Bodegas, G; Prieto, J C; Arilla, E

    1987-04-01

    Administration of cysteamine in rabbits elicited a rapid depletion of both duodenal mucosa and plasma somatostatin. A significant reduction was observed within 5 min, returning toward control values by 150 min. The depletion of somatostatin was associated with an increase in the binding capacity and a decrease in the affinity of both high- and low-affinity binding sites present in cytosol of duodenal mucosa. Incubation of cytosolic fraction from control rabbits with 1 mM cysteamine did not modify somatostatin binding. Furthermore, addition of cysteamine at the time of binding assay did not affect the integrity of 125I-Tyr11-somatostatin. It is concluded that in vivo administration of cysteamine to rabbits depletes both duodenal mucosa and plasma somatostatin and leads to up-regulation of duodenal somatostatin binding sites.

  7. Colloidal Plasmas : Basic physics of colloidal plasmas

    Indian Academy of Sciences (India)

    C B Dwivedi

    2000-11-01

    Colloidal plasma is a distinct class of the impure plasmas with multispecies ionic composition. The distinction lies in the phase distribution of the impurity-ion species. The ability to tailor the electrostatic interactions between these colloidal particles provides a fertile ground for scientists to investigate the fundamental aspects of the Coulomb phase transition behavior. The present contribution will review the basic physics of the charging mechanism of the colloidal particles as well as the physics of the collective normal mode behavior of the general multi-ion species plasmas. Emphasis will be laid on the clarification of the prevailing confusing ideas about distinct qualities of the various acoustic modes, which are likely to exist in colloidal plasmas as well as in normal multi-ion species plasmas. Introductory ideas about the proposed physical models for the Coulomb phase transition in colloidal plasma will also be discussed.

  8. Stability of the transthyretin molecule as a key factor in the interaction with a-beta peptide--relevance in Alzheimer's disease.

    Directory of Open Access Journals (Sweden)

    Carlos A Ribeiro

    Full Text Available Transthyretin (TTR protects against A-Beta toxicity by binding the peptide thus inhibiting its aggregation. Previous work showed different TTR mutations interact differently with A-Beta, with increasing affinities correlating with decreasing amyloidogenecity of the TTR mutant; this did not impact on the levels of inhibition of A-Beta aggregation, as assessed by transmission electron microscopy. Our work aimed at probing differences in binding to A-Beta by WT, T119M and L55P TTR using quantitative assays, and at identifying factors affecting this interaction. We addressed the impact of such factors in TTR ability to degrade A-Beta. Using a dot blot approach with the anti-oligomeric antibody A11, we showed that A-Beta formed oligomers transiently, indicating aggregation and fibril formation, whereas in the presence of WT and T119M TTR the oligomers persisted longer, indicative that these variants avoided further aggregation into fibrils. In contrast, L55PTTR was not able to inhibit oligomerization or to prevent evolution to aggregates and fibrils. Furthermore, apoptosis assessment showed WT and T119M TTR were able to protect against A-Beta toxicity. Because the amyloidogenic potential of TTR is inversely correlated with its stability, the use of drugs able to stabilize TTR tetrameric fold could result in increased TTR/A-Beta binding. Here we showed that iododiflunisal, 3-dinitrophenol, resveratrol, [2-(3,5-dichlorophenylamino] (DCPA and [4-(3,5-difluorophenyl] (DFPB were able to increase TTR binding to A-Beta; however only DCPA and DFPB improved TTR proteolytic activity. Thyroxine, a TTR ligand, did not influence TTR/A-Beta interaction and A-Beta degradation by TTR, whereas RBP, another TTR ligand, not only obstructed the interaction but also inhibited TTR proteolytic activity. Our results showed differences between WT and T119M TTR, and L55PTTR mutant regarding their interaction with A-Beta and prompt the stability of TTR as a key factor in this

  9. Synaptotagmin-1 binds to PIP(2)-containing membrane but not to SNAREs at physiological ionic strength.

    Science.gov (United States)

    Park, Yongsoo; Seo, Jong Bae; Fraind, Alicia; Pérez-Lara, Angel; Yavuz, Halenur; Han, Kyungreem; Jung, Seung-Ryoung; Kattan, Iman; Walla, Peter Jomo; Choi, MooYoung; Cafiso, David S; Koh, Duk-Su; Jahn, Reinhard

    2015-10-01

    The Ca(2+) sensor synaptotagmin-1 is thought to trigger membrane fusion by binding to acidic membrane lipids and SNARE proteins. Previous work has shown that binding is mediated by electrostatic interactions that are sensitive to the ionic environment. However, the influence of divalent or polyvalent ions, at physiological concentrations, on synaptotagmin's binding to membranes or SNAREs has not been explored. Here we show that binding of rat synaptotagmin-1 to membranes containing phosphatidylinositol 4,5-bisphosphate (PIP2) is regulated by charge shielding caused by the presence of divalent cations. Surprisingly, polyvalent ions such as ATP and Mg(2+) completely abrogate synaptotagmin-1 binding to SNAREs regardless of the presence of Ca(2+). Altogether, our data indicate that at physiological ion concentrations Ca(2+)-dependent synaptotagmin-1 binding is confined to PIP2-containing membrane patches in the plasma membrane, suggesting that membrane interaction of synaptotagmin-1 rather than SNARE binding triggers exocytosis of vesicles.

  10. Binding Principles A and B

    Institute of Scientific and Technical Information of China (English)

    陈源

    2014-01-01

    This paper focuses on the discussion of how Binding Principle A and Binding Principe B help with the interpretation of reference in English and Chinese. They are supposedly universal across languages.

  11. Identification of actin binding protein, ABP-280, as a binding partner of human Lnk adaptor protein.

    Science.gov (United States)

    He, X; Li, Y; Schembri-King, J; Jakes, S; Hayashi, J

    2000-08-01

    Human Lnk (hLnk) is an adaptor protein with multiple functional domains that regulates T cell activation signaling. In order to identify cellular Lnk binding partners, a yeast two-hybrid screening of human spleen cDNA library was carried out using human hLnk as bait. A polypeptide sequence identical to the C-terminal segment of the actin binding protein (ABP-280) was identified as a hLnk binding protein. The expressed hLnk and the FLAG tagged C-terminal 673 amino acid residues of ABP-280 or the endogenous ABP-280 in COS-7 cells could be co-immunoprecipitated using antibodies either to hLnk, FLAG or ABP-280, respectively. Furthermore, immunofluorescence confocal microscope showed that hLnk and ABP-280 co-localized at the plasma membrane and at juxtanuclear region of COS-7 cells. In Jurkat cells, the endogenous hLnk also associates with the endogenous ABP-280 indicating that the association of these two proteins is physiological. The interacting domains of both proteins were mapped using yeast two-hybrid assays. Our results indicate that hLnk binds to the residues 2006-2454 (repeats 19-23C) of ABP-280. The domain in hLnk that associates with ABP-280 was mapped to an interdomain region of 56 amino acids between pleckstrin homology and Src homology 2 domains. These results suggest that hLnk may exert its regulatory role through its association with ABP-280.

  12. Nonlinear plasma wave in magnetized plasmas

    Energy Technology Data Exchange (ETDEWEB)

    Bulanov, Sergei V. [Kansai Photon Science Institute, JAEA, Kizugawa, Kyoto 619-0215 (Japan); Prokhorov Institute of General Physics, Russian Academy of Sciences, Moscow 119991 (Russian Federation); Moscow Institute of Physics and Technology, Dolgoprudny, Moscow region 141700 (Russian Federation); Esirkepov, Timur Zh.; Kando, Masaki; Koga, James K. [Kansai Photon Science Institute, JAEA, Kizugawa, Kyoto 619-0215 (Japan); Hosokai, Tomonao; Zhidkov, Alexei G. [Photon Pioneers Center, Osaka University, 2-8 Yamadaoka, Suita, Osaka 565-0871 (Japan); Japan Science and Technology Agency, CREST, 2-1, Yamadaoka, Suita, Osaka 565-0871 (Japan); Kodama, Ryosuke [Photon Pioneers Center, Osaka University, 2-8 Yamadaoka, Suita, Osaka 565-0871 (Japan); Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871 (Japan)

    2013-08-15

    Nonlinear axisymmetric cylindrical plasma oscillations in magnetized collisionless plasmas are a model for the electron fluid collapse on the axis behind an ultrashort relativisically intense laser pulse exciting a plasma wake wave. We present an analytical description of the strongly nonlinear oscillations showing that the magnetic field prevents closing of the cavity formed behind the laser pulse. This effect is demonstrated with 3D PIC simulations of the laser-plasma interaction. An analysis of the betatron oscillations of fast electrons in the presence of the magnetic field reveals a characteristic “Four-Ray Star” pattern.

  13. Elements of plasma technology

    CERN Document Server

    Wong, Chiow San

    2016-01-01

    This book presents some fundamental aspects of plasma technology that are important for beginners interested to start research in the area of plasma technology . These include the properties of plasma, methods of plasma generation and basic plasma diagnostic techniques. It also discusses several low cost plasma devices, including pulsed plasma sources such as plasma focus, pulsed capillary discharge, vacuum spark and exploding wire; as well as low temperature plasmas such as glow discharge and dielectric barrier discharge which the authors believe may have potential applications in industry. The treatments are experimental rather than theoretical, although some theoretical background is provided where appropriate. The principles of operation of these devices are also reviewed and discussed.

  14. Stability, protein binding and clearance studies of [99mTc]DTPA. Evaluation of a commercially available dry-kit

    DEFF Research Database (Denmark)

    Rehling, M

    1988-01-01

    [99mTc]DTPA has achieved widespread use for the measurement of glomerular filtration rate (GFR) with the single injection plasma clearance technique and for gamma-camera renography. However, the quality of the commercial preparations varies. The purpose of the present investigation was to study...... the quality of a commercial [99mTc]DTPA preparation (C.I.S., France) with reference to stability, protein binding and accuracy of the determined plasma clearance values as a measure of GFR. The stability of the preparations was studied by thin-layer chromatography, the in vitro protein binding by Sephadex...... filtration after incubation with human serum albumin and in vivo protein binding by filtration of human plasma. The accuracy of the plasma clearance values was investigated by comparison with the simultaneously measured plasma clearance of [51Cr]EDTA. There was no detectable free pertechnetate or hydrolysed...

  15. Free energy calculations on Transthyretin dissociation and ligand binding from Molecular Dynamics Simulations

    DEFF Research Database (Denmark)

    Sørensen, Jesper; Hamelberg, Donald; McCammon, J. Andrew

    experimental results have helped to explain this aberrant behavior of TTR, however, structural insights of the amyloidgenic process are still lacking. Therefore, we have used all-atom molecular dynamics simulation and free energy calculations to study the initial phase of this process. We have calculated...... the free energy changes of the initial tetramer dissociation under different conditions and in the presence of thyroxine....

  16. Free energy calculations on Transthyretin dissociation and ligand binding from Molecular Dynamics Simulations

    DEFF Research Database (Denmark)

    Sørensen, Jesper; Hamelberg, Donald; McCammon, J. Andrew

    experimental results have helped to explain this aberrant behavior of TTR, however, structural insights of the amyloidgenic process are still lacking. Therefore, we have used all-atom molecular dynamics simulation and free energy calculations to study the initial phase of this process. We have calculated...... the free energy changes of the initial tetramer dissociation under different conditions and in the presence of thyroxine....

  17. Reviews of plasma physics

    CERN Document Server

    2008-01-01

    "Reviews of Plasma Physics Volume 24," edited by V.D. Shafranov, presents two reviews from the cutting-edge of Russian plasma physics research. The first review by V.A. Rozhansky devoted to the mechanisms of transverse conductivity and generation of self-consistent electric fields in strongly ionized magnetized plasma. The second review by O.G. Bakunin considers numerous aspects of turbulent transport in plasma and fluids. This review is focused on scaling arguments for describing anomalous diffusion in the presence of complex structures. These topics are especially important for fusion plasma research, plasma astrophysics, discharge physics, and turbulence

  18. Reviews of plasma physics

    Energy Technology Data Exchange (ETDEWEB)

    Shafranov, Vitalii Dmitrievich (ed.); Bakunin, Oleg G. (comps.) [Rossijskij Nauchnyj Tsentr ' ' Kurchatovskij Inst.' ' , Moscow (Russian Federation). Nuclear Fusion Inst.; Rozhansky, V. [St. Petersburg State Polytechnical Univ. (Russian Federation)

    2008-07-01

    Reviews of Plasma Physics Volume 24, edited by V.D. Shafranov, presents two reviews from the cutting-edge of Russian plasma physics research. The first review by V.A. Rozhansky devoted to the mechanisms of transverse conductivity and generation of self-consistent electric fields in strongly ionized magnetized plasma. The second review by O.G. Bakunin considers numerous aspects of turbulent transport in plasma and fluids. This review is focused on scaling arguments for describing anomalous diffusion in the presence of complex structures. These topics are especially important for fusion plasma research, plasma astrophysics, discharge physics, and turbulence (orig.)

  19. Carboplatin binding to histidine

    Energy Technology Data Exchange (ETDEWEB)

    Tanley, Simon W. M. [University of Manchester, Brunswick Street, Manchester M13 9PL (United Kingdom); Diederichs, Kay [University of Konstanz, D-78457 Konstanz (Germany); Kroon-Batenburg, Loes M. J. [Utrecht University, Padualaan 8, 3584 CH Utrecht (Netherlands); Levy, Colin [University of Manchester, 131 Princess Street, Manchester M1 7DN (United Kingdom); Schreurs, Antoine M. M. [Utrecht University, Padualaan 8, 3584 CH Utrecht (Netherlands); Helliwell, John R., E-mail: john.helliwell@manchester.ac.uk [University of Manchester, Brunswick Street, Manchester M13 9PL (United Kingdom)

    2014-08-29

    An X-ray crystal structure showing the binding of purely carboplatin to histidine in a model protein has finally been obtained. This required extensive crystallization trials and various novel crystal structure analyses. Carboplatin is a second-generation platinum anticancer agent used for the treatment of a variety of cancers. Previous X-ray crystallographic studies of carboplatin binding to histidine (in hen egg-white lysozyme; HEWL) showed the partial conversion of carboplatin to cisplatin owing to the high NaCl concentration used in the crystallization conditions. HEWL co-crystallizations with carboplatin in NaBr conditions have now been carried out to confirm whether carboplatin converts to the bromine form and whether this takes place in a similar way to the partial conversion of carboplatin to cisplatin observed previously in NaCl conditions. Here, it is reported that a partial chemical transformation takes place but to a transplatin form. Thus, to attempt to resolve purely carboplatin binding at histidine, this study utilized co-crystallization of HEWL with carboplatin without NaCl to eliminate the partial chemical conversion of carboplatin. Tetragonal HEWL crystals co-crystallized with carboplatin were successfully obtained in four different conditions, each at a different pH value. The structural results obtained show carboplatin bound to either one or both of the N atoms of His15 of HEWL, and this particular variation was dependent on the concentration of anions in the crystallization mixture and the elapsed time, as well as the pH used. The structural details of the bound carboplatin molecule also differed between them. Overall, the most detailed crystal structure showed the majority of the carboplatin atoms bound to the platinum centre; however, the four-carbon ring structure of the cyclobutanedicarboxylate moiety (CBDC) remained elusive. The potential impact of the results for the administration of carboplatin as an anticancer agent are described.

  20. Sperm-Egg Interaction: Evidence for Boar Sperm Plasma Membrane Receptors for Porcine Zona Pellucida

    Science.gov (United States)

    Peterson, Rudolph N.; Russell, Lonnie; Bundman, Donna; Freund, Matthew

    1980-01-01

    Freshly ejaculated, noncapacitated boar sperm bind rapidly and in large numbers to pig egg zona pellucida in vitro. In the present study, the number of sperm bound decreased sharply when sperm motility was lowered by energy poisons or by reducing the temperature. Highly motile sperm from humans, guinea pigs, and rats, added at concentrations ten times higher than control sperm, did not bind to the porcine zona. At the same high concentration, a small number of hamster and bull sperm bound to the zona. Binding of boar sperm to the zona pellucida was blocked almost completely by diluted whole antiserum to sperm plasma membranes and by univalent (Fab) antibody to these membranes. When antibody to sperm plasma membrane was first absorbed with plasma membrane vesicles, sperm binding was not inhibited. These results provide direct evidence for the existence of sperm plasma membrane receptors for the zona pellucida of the pig.

  1. Does protein binding modulate the effect of angiotensin II receptor antagonists?

    Directory of Open Access Journals (Sweden)

    Marc P Maillard

    2001-03-01

    Full Text Available IntroductionAngiotensin II AT 1-receptor antagonists are highly bound to plasma proteins (≥ 99%. With some antagonists, such as DuP-532, the protein binding was such that no efficacy of the drug could be demonstrated clinically. Whether protein binding interferes with the efficacy of other antagonists is not known. We have therefore investigated in vitro how plasma proteins may affect the antagonistic effect of different AT1-receptor antagonists.MethodsA radio-receptor binding assay was used to analyse the interaction between proteins and the ability of various angiotensin II (Ang II antagonists to block AT1-receptors. In addition, the Biacore technology, a new technique which enables the real-time monitoring of binding events between two molecules, was used to evaluate the dissociation rate constants of five AT1-receptor antagonists from human serum albumin.ResultsThe in vitro AT 1-antagonistic effects of different Ang II receptor antagonists were differentially affected by the presence of human plasma, with rightward shifts of the IC50 ranging from one to several orders of magnitude. The importance of the shift correlates with the dissociation rate constants of these drugs from albumin. Our experiments also show that the way that AT1-receptor antagonists bind to proteins differs from one compound to another. These results suggest that the interaction with plasma proteins appears to modulate the efficacy of some Ang II antagonists.ConclusionAlthough the high binding level of Ang II receptor antagonist to plasma proteins appears to be a feature common to this class of compounds, the kinetics and characteristics of this binding is of great importance. With some antagonists, protein binding interferes markedly with their efficacy to block AT1-receptors.

  2. Evaluation of binding strength depending on the adhesive binding methods

    Directory of Open Access Journals (Sweden)

    Suzana Pasanec Preprotić

    2015-05-01

    Full Text Available A book with a personal value is worth remembering since it represents specific interests of an individual - author of the book. Therefore the original is the first issue of a book which is always bound manually. Due to cost-effectiveness, adhesive binding is most commonly used in author’s edition in paperback and hardback. Adhesive binding methods differ only if a paper leaf is a binding unit in adhesive binding form. The subject of the research is the quality of book block binding for two binding methods with/without mull fabric. The assumption is that double-fan adhesive binding method shows an extraordinary binding quality as compared to the rough spine method. For the needs of this research book block parameters remained unaltered: paper type, size and book volume. The results related to strength were obtained by using an experimental method of tensile strength for individual paper leaves. The rating of book block quality was conducted in accordance with FOGRA Nr.71006 guidelines for page pull-test. Furthermore, strength results for both methods were compared in order to evaluate the importance of changing the quality of adhesive binding. Statistical method ANOVA analysis of variance and Fisher’s F-test were used to evaluate the quality of book block binding.

  3. Clinical manifestations of mannan-binding lectin deficiency

    DEFF Research Database (Denmark)

    Thiel, Steffen; Frederiksen, Pernille Dorthea; Jensenius, Jens Christian

    2006-01-01

    Mannan-binding lectin (MBL) is a plasma protein of the innate immune system with the ability to initiate antimicrobial and inflammatory actions. MBL deficiency is common. More than 10% of the general population may, depending on definition, be classified as MBL deficient, underlining the redundan...... of the immune system. Ongoing research attempt to illuminate at which conditions MBL deficiency may lead to disease. With examples, this review illustrates the diversity of results obtained so far....

  4. Analysis of Hydrophilic Mercury-Binding Proteins in Medaka (Oryzias latipes) Using High Performance Liquid Chromatography Coupled with Inductively Coupled Plasma-Mass Spectrometry%高效液相色谱-电感耦合等离子体质谱研究Medaka体内水溶性汞结合蛋白

    Institute of Scientific and Technical Information of China (English)

    李璐; 何滨; 江桂斌

    2011-01-01

    The hydrophilic mercury-binding proteins in the brain and liver extractions of Medaka after methylmercury and inorganic mercury exposure were analyzed by high performance liquid chromatography coupled with inductively coupled plasma mass spectrometry. Size exclusion chromatography was selected in the experiment with a mobile phase of 0. 15 mol/L NHaAc, running at a 0.5 mL/min flow rate. Concentrations of total copper, zinc and mercury in the brain and liver extracts of Medaka were determined by ICP-MS. Detection limits for these three elements were in the range of 0.01 μg/L to 0.05 μg/L, and the spike recoveries ranged from 93. 1%-97. 0%. Compared with the control group, high level of mercury is accumulated in the brain and liver of exposure groups. The distribution patterns of mercury-containing proteins in the brain and liver extracts of Medaka between methylmercury and inorganic mercury exposure group were different. The concentrations of copper and zinc in the brain and liver extractions of both exposure groups were decreased. The studies of the mercurycontaining protein fractions in Medaka are helpful for the understanding of the mechanisms of mercury toxicity.%采用高效液相色谱与电感耦合等离子体质谱联用技术,对不同形态汞暴露后的Medaka肝脏和脑组织中水溶性蛋白进行研究,以体积排阻柱为液相分离柱,流动相为0.15 mol/L NH4Ac,流速为0.5 mL/min.利用电感耦合等离子体质谱对Medaka肝脏和脑组织提取液中铜、锌和汞的含量进行了分析,检出限为0.01~0.05 μg/L,元素的加标回收率为93.1%~97.0%.通过与空白组对比发现,汞暴露后的Medaka鱼体肝脏和脑组织对水体中的汞有一定程度的富集;不同形态汞暴露后,在Medaka肝脏和脑组织中的分布明显不同,并且都会在一定程度上降低Medaka肝脏和脑组织中铜与锌的含量,有利于进一步研究汞在生物体内的毒性机理.

  5. A role for carbohydrate recognition in mammalian sperm-egg binding

    Energy Technology Data Exchange (ETDEWEB)

    Clark, Gary F., E-mail: clarkgf@health.missouri.edu

    2014-08-01

    Highlights: • Mammalian sperm-egg binding as a carbohydrate dependent species recognition event. • The role of carbohydrate recognition in human, mouse and pig sperm-egg binding. • Historical perspective and future directions for research focused on gamete binding. - Abstract: Mammalian fertilization usually requires three sequential cell–cell interactions: (i) initial binding of sperm to the specialized extracellular matrix coating the egg known as the zona pellucida (ZP); (ii) binding of sperm to the ZP via the inner acrosomal membrane that is exposed following the induction of acrosomal exocytosis; and (iii) adhesion of acrosome-reacted sperm to the plasma membrane of the egg cell, enabling subsequent fusion of these gametes. The focus of this review is on the initial binding of intact sperm to the mammalian ZP. Evidence collected over the past fifty years has confirmed that this interaction relies primarily on the recognition of carbohydrate sequences presented on the ZP by lectin-like egg binding proteins located on the plasma membrane of sperm. There is also evidence that the same carbohydrate sequences that mediate binding also function as ligands for lectins on lymphocytes that can inactivate immune responses, likely protecting the egg and the developing embryo up to the stage of blastocyst hatching. The literature related to initial sperm-ZP binding in the three major mammalian models (human, mouse and pig) is discussed. Historical perspectives and future directions for research related to this aspect of gamete adhesion are also presented.

  6. Hot plasma dielectric tensor

    NARCIS (Netherlands)

    Westerhof, E.

    1996-01-01

    The hot plasma dielectric tensor is discussed in its various approximations. Collisionless cyclotron resonant damping and ion/electron Bernstein waves are discussed to exemplify the significance of a kinetic description of plasma waves.

  7. Special issue: Plasma Conversion

    NARCIS (Netherlands)

    Nozaki, T.; Bogaerts, A.; Tu, X.; van de Sanden, M. C. M.

    2017-01-01

    With growing concern of energy and environmental issues, the combination of plasma and heterogeneous catalysts receives special attention in greenhouse gas conversion, nitrogen fixation and hydrocarbon chemistry. Plasma gas conversion driven by renewable electricity is particularly important for the

  8. Treponema pallidum receptor binding proteins interact with fibronectin

    Energy Technology Data Exchange (ETDEWEB)

    Peterson, K.M.; Baseman, J.B.; Alderete, J.F.

    1983-06-01

    Analysis of plasma proteins avidly bound to T. pallidum surfaces revealed the ability of T. pallidum to acquire numerous host macromolecules. No acquisition was evident by the avirulent spirochete, T. phagedenis biotype Reiter. Western blotting technology using hyperimmune antifibronectin serum as a probe revealed the ability of virulent treponemes to avidly bind fibronectin from a complex medium such as plasma. The specificity of the tiplike adherence of motile T. pallidum to fibronectin-coated glass surfaces and to fibronectin on HEp-2 cells was reinforced by the observation that pretreatment of coverslips or cell monolayers with monospecific antiserum against fibronectin substantially reduced T. pallidum attachment. The stoichiometric binding of T. pallidum to fibronectin-coated coverslips and the inability of unlabeled or /sup 35/S-radiolabeled treponemes to interact with glass surfaces treated with other plasma proteins further established the specific nature of the interaction between virulent T. pallidum and fibronectin. The avid association between three outer envelope proteins of T. pallidum and fibronectin was also demonstrated. These treponemal surface proteins have been previously identified as putative receptor-binding proteins responsible for T. pallidum parasitism of host cells. The data suggest that surface fibronectin mediates tip-oriented attachment of T. pallidum to host cells via a receptor-ligand mechanism of recognition.

  9. Microwave Argon Plasma Torch

    Science.gov (United States)

    2013-07-01

    an electron-ion pair in the discharge. Fig. 2. EEDF is non - Maxwellian and changes along the plasma column The electron–neutral collision...plasma radius. Even at atmospheric pressure the EEDF is non - Maxwellian and it is changing along the plasma column. ...18 31st ICPIG, July 14-19, 2013, Granada, Spain EEDF usually strongly differs from Maxwellian and chages along the plasma column (this is

  10. Introduction to Plasma Physics

    Science.gov (United States)

    Gurnett, Donald A.; Bhattacharjee, Amitava

    2017-03-01

    Preface; 1. Introduction; 2. Characteristic parameters of a plasma; 3. Single particle motions; 4. Waves in a cold plasma; 5. Kinetic theory and the moment equations; 6. Magnetohydrodynamics; 7. MHD equilibria and stability; 8. Discontinuities and shock waves; 9. Electrostatic waves in a hot unmagnetized plasma; 10. Waves in a hot magnetized plasma; 11. Nonlinear effects; 12. Collisional processes; Appendix A. Symbols; Appendix B. Useful trigonometric identities; Appendix C. Vector differential operators; Appendix D. Vector calculus identities; Index.

  11. Plasma physics an introduction

    CERN Document Server

    Fitzpatrick, Richard

    2014-01-01

    Plasma Physics: An Introduction is based on a series of university course lectures by a leading name in the field, and thoroughly covers the physics of the fourth state of matter. This book looks at non-relativistic, fully ionized, nondegenerate, quasi-neutral, and weakly coupled plasma. Intended for the student market, the text provides a concise and cohesive introduction to plasma physics theory, and offers a solid foundation for students wishing to take higher level courses in plasma physics.

  12. The chaperone and potential mannan-binding lectin (MBL) co-receptor calreticulin interacts with MBL through the binding site for MBL-associated serine proteases.

    Science.gov (United States)

    Pagh, Rasmus; Duus, Karen; Laursen, Inga; Hansen, Paul R; Mangor, Julie; Thielens, Nicole; Arlaud, Gérard J; Kongerslev, Leif; Højrup, Peter; Houen, Gunnar

    2008-02-01

    The chaperone calreticulin has been suggested to function as a C1q and collectin receptor. The interaction of calreticulin with mannan-binding lectin (MBL) was investigated by solid-phase binding assays. Calreticulin showed saturable and time-dependent binding to recombinant MBL, provided that MBL was immobilized on a solid surface or bound to mannan on a surface. The binding was non-covalent and biphasic with an initial salt-sensitive phase followed by a more stable salt-insensitive interaction. For plasma-derived MBL, known to be complexed with MBL-associated serine proteases (MASPs), no binding was observed. Interaction of calreticulin with recombinant MBL was fully inhibited by recombinant MASP-2, MASP-3 and MAp19, but not by the MASP-2 D105G and MAp19 Y59A variants characterized by defective MBL binding ability. Furthermore, MBL point mutants with impaired MASP binding showed no interaction with calreticulin. Comparative analysis of MBL with complement component C1q, its counterpart of the classical pathway, revealed that they display similar binding characteristics for calreticulin, providing further indication that calreticulin is a common co-receptor/chaperone for both proteins. In conclusion, the potential MBL co-receptor calreticulin binds to MBL at the MASP binding site and the interaction may involve a conformational change in MBL.

  13. Lipids and lipid binding proteins: a perfect match.

    Science.gov (United States)

    Glatz, Jan F C

    2015-02-01

    Lipids serve a great variety of functions, ranging from structural components of biological membranes to signaling molecules affecting various cellular functions. Several of these functions are related to the unique physico-chemical properties shared by all lipid species, i.e., their hydrophobicity. The latter, however, is accompanied by a poor solubility in an aqueous environment and thus a severe limitation in the transport of lipids in aqueous compartments such as blood plasma and the cellular soluble cytoplasm. Specific proteins which can reversibly and non-covalently associate with lipids, designated as lipid binding proteins or lipid chaperones, greatly enhance the aqueous solubility of lipids and facilitate their transport between tissues and within tissue cells. Importantly, transport of lipids across biological membranes also is facilitated by specific (membrane-associated) lipid binding proteins. Together, these lipid binding proteins determine the bio-availability of their ligands, and thereby markedly influence the subsequent processing, utilization, or signaling effect of lipids. The bio-availability of specific lipid species thus is governed by the presence of specific lipid binding proteins, the affinity of these proteins for distinct lipid species, and the presence of competing ligands (including pharmaceutical compounds). Recent studies suggest that post-translational modifications of lipid binding proteins may have great impact on lipid-protein interactions. As a result, several levels of regulation exist that together determine the bio-availability of lipid species. This short review discusses the significance of lipid binding proteins and their potential application as targets for therapeutic intervention.

  14. Pleiotropic virulence factor - Streptococcus pyogenes fibronectin-binding proteins.

    Science.gov (United States)

    Yamaguchi, Masaya; Terao, Yutaka; Kawabata, Shigetada

    2013-04-01

    Streptococcus pyogenes causes a broad spectrum of infectious diseases, including pharyngitis, skin infections and invasive necrotizing fasciitis. The initial phase of infection involves colonization, followed by intimate contact with the host cells, thus promoting bacterial uptake by them. S. pyogenes recognizes fibronectin (Fn) through its own Fn-binding proteins to obtain access to epithelial and endothelial cells in host tissue. Fn-binding proteins bind to Fn to form a bridge to α5 β1 -integrins, which leads to rearrangement of cytoskeletal actin in host cells and uptake of invading S. pyogenes. Recently, several structural analyses of the invasion mechanism showed molecular interactions by which Fn converts from a compact plasma protein to a fibrillar component of the extracellular matrix. After colonization, S. pyogenes must evade the host innate immune system to spread into blood vessels and deeper organs. Some Fn-binding proteins contribute to evasion of host innate immunity, such as the complement system and phagocytosis. In addition, Fn-binding proteins have received focus as non-M protein vaccine candidates, because of their localization and conservation among different M serotypes.Here, we review the roles of Fn-binding proteins in the pathogenesis and speculate regarding possible vaccine antigen candidates. © 2012 Blackwell Publishing Ltd.

  15. Plasma membrane ATPases

    DEFF Research Database (Denmark)

    Palmgren, Michael Broberg; Bækgaard, Lone; Lopez Marques, Rosa Laura

    2011-01-01

    The plasma membrane separates the cellular contents from the surrounding environment. Nutrients must enter through the plasma membrane in order to reach the cell interior, and toxic metabolites and several ions leave the cell by traveling across the same barrier. Biological pumps in the plasma me...

  16. Plasma in dentistry

    OpenAIRE

    Cha, Seunghee; Park, Young-Seok

    2014-01-01

    This review describes the contemporary aspects of plasma application in dentistry. Previous studies on plasma applications were classified into two categories, surface treatment and direct applications, and were reviewed, respectively according to the approach. The current review discussed modification of dental implant surface, enhancing of adhesive qualities, enhancing of polymerization, surface coating and plasma cleaning under the topics of surface treatment. Microbicidal activities, deco...

  17. The Plasma Universe

    Science.gov (United States)

    Suplee, Curt

    2009-09-01

    Preface; 1. The fourth state of matter; 2. The music and dance of plasmas; 3. The Sun-Earth connection; 4. Bringing the Sun to Earth: the story of controlled thermonuclear fusion; 5. The cosmic plasma theater: galaxies, stars, and accretion disks; 6. Putting plasmas to work; Index.

  18. Kinetics characterization of c-Src binding to lipid membranes: Switching from labile to persistent binding.

    Science.gov (United States)

    Le Roux, Anabel-Lise; Busquets, Maria Antònia; Sagués, Francesc; Pons, Miquel

    2016-02-01

    Cell signaling by the c-Src proto-oncogen requires the attachment of the protein to the inner side of the plasma membrane through the myristoylated N-terminal region, known as the SH4 domain. Additional binding regions of lower affinity are located in the neighbor intrinsically disordered Unique domain and the structured SH3 domain. Here we present a surface plasmon resonance study of the binding of a myristoylated protein including the SH4, Unique and SH3 domains of c-Src to immobilized liposomes. Two distinct binding processes were observed: a fast and a slow one. The second process lead to a persistently bound form (PB) with a slower binding and a much slower dissociation rate than the first one. The association and dissociation of the PB form could be detected using an anti-SH4 antibody. The kinetic analysis revealed that binding of the PB form follows a second order rate law suggesting that it involves the formation of c-Src dimers on the membrane surface. A kinetically equivalent PB form is observed in a myristoylated peptide containing only the SH4 domain but not in a construct including the three domains but with a 12-carbon lauroyl substituent instead of the 14-carbon myristoyl group. The PB form is observed with neutral lipids but its population increases when the immobilized liposomes contain negatively charged lipids. We suggest that the PB form may represent the active signaling form of c-Src while the labile form provides the capacity for fast 2D search of the target signaling site on the membrane surface.

  19. Níveis plasmáticos de vitamina A, carotenóides e proteína ligadora de retinol em crianças com infecções respiratórias agudas e doenças diarréicas Plasma levels of vitamin A, carotenoids and retinol binding protein in children with acute respiratory infections and diarrhoeal diseases

    Directory of Open Access Journals (Sweden)

    Gustavo Velasquez-Melendez

    1994-10-01

    Full Text Available Planejou-se um estudo com o objetivo de se avaliar os níveis plasmáticos de vitamina A, carotenóides e proteína ligadora de retinol (RBP em 311 crianças, de 7 meses a onze anos de idade, com história de infecções das vias aéreas superiores (IVAS, pneumonia e diarréia, residentes na área urbana da Cidade de São Paulo, Brasil, e atendidas no serviço de pediatria de um hospital-escola. As dosagens de vitamina A e carotenóides realizaram-se pelo método de Neeld-Pearson e o RBP pelo método de Mancini. Os níveis plasmáticos de vitamina A (µg/dl e RBP (mg/dl foram mais baixos (pThe present study was carried out in order to assess the plasma levels of vitamin A, carotenoids and retinol binding protein (RBP of three-hundred and eleven children aged from seven months to eleven years, who had a history of upper respiratory infection (URI, pneumonia and diarrhoea. The children were resident in the urban area of the Municipality of S. Paulo, Brazil, and were seen at the pediatric service of the one school-hospital. The data show that plasma vitamin A (µg/dl and RBP (mg/ dl levels in the diarrhoea (15.2 µg/dl; 1.7 mg/dl and pneumonia (15.2 µg/dl; 0.7 mg/dl groups were lower (p<0.05 than those observed in the control (18.8 µg/dl; 2.6 mg/dl and URI (19.0 µg/dl; 2.4 mg/dl groups. The plasma carotenoid levels were lower in all groups than in the control group (p<0.05. These findings corroborate the results that show low levels of vitamin A in circulation during period of infection.

  20. Stability, protein binding and clearance studies of [99mTc]DTPA. Evaluation of a commercially available dry-kit

    DEFF Research Database (Denmark)

    Rehling, M

    1988-01-01

    the quality of a commercial [99mTc]DTPA preparation (C.I.S., France) with reference to stability, protein binding and accuracy of the determined plasma clearance values as a measure of GFR. The stability of the preparations was studied by thin-layer chromatography, the in vitro protein binding by Sephadex...

  1. Analytic QCD Binding Potentials

    CERN Document Server

    Fried, H M; Grandou, T; Sheu, Y -M

    2011-01-01

    This paper applies the analytic forms of a recent non-perturbative, manifestly gauge- and Lorentz-invariant description (of the exchange of all possible virtual gluons between quarks ($Q$) and/or anti-quarks ($\\bar{Q}$) in a quenched, eikonal approximation) to extract analytic forms for the binding potentials generating a model $Q$-$\\bar{Q}$ "pion", and a model $QQQ$ "nucleon". Other, more complicated $Q$, $\\bar{Q}$ contributions to such color-singlet states may also be identified analytically. An elementary minimization technique, relevant to the ground states of such bound systems, is adopted to approximate the solutions to a more proper, but far more complicated Schroedinger/Dirac equation; the existence of possible contributions to the pion and nucleon masses due to spin, angular momentum, and "deformation" degrees of freedom is noted but not pursued. Neglecting electromagnetic and weak interactions, this analysis illustrates how the one new parameter making its appearance in this exact, realistic formali...

  2. The chaperone and potential mannan-binding lectin (MBL) co-receptor calreticulin interacts with MBL through the binding site for MBL-associated serine proteases

    DEFF Research Database (Denmark)

    Pagh, Rasmus; Duus, Karen; Laursen, Inga;

    2008-01-01

    was immobilized on a solid surface or bound to mannan on a surface. The binding was non-covalent and biphasic with an initial salt-sensitive phase followed by a more stable salt-insensitive interaction. For plasma-derived MBL, known to be complexed with MBL-associated serine proteases (MASPs), no binding...... with calreticulin. Comparative analysis of MBL with complement component C1q, its counterpart of the classical pathway, revealed that they display similar binding characteristics for calreticulin, providing further indication that calreticulin is a common co-receptor/chaperone for both proteins. In conclusion...

  3. Cyclotron waves in plasma

    CERN Document Server

    Lominadze, D G

    2013-01-01

    Cyclotron Waves in Plasma is a four-chapter text that covers the basic physical concepts of the theory of cyclotron waves and cyclotron instabilities, brought about by the existence of steady or alternating plasma currents flowing perpendicular to the magnetic field.This book considers first a wide range of questions associated with the linear theory of cyclotron oscillations in equilibrium plasmas and in electron plasmas in metals and semiconductors. The next chapter deals with the parametric excitation of electron cyclotron oscillations in plasma in an alternating electric field. A chapter f

  4. Plasma Therapy: An Overview

    Directory of Open Access Journals (Sweden)

    Rajkumar Diwan

    2011-01-01

    Full Text Available Definition: Plasma, the fourth state of matter, is a collection of charged particles (electrons, ions, neutral atoms. Recent demonstration of plasma technology in treatment of living cells, tissue and organs are creating a new field at the intersection of plasma science and technology with biology and medicine known as plasma medicine. Plasma medicine is one of the newest fields of modem applied plasma chemistry. It appeared several years ago and comprises studies concerning the direct action of low-temperature, one atmosphere air plasma (cold plasma/nonthermal plasmalnonequilibrium on body tissues for various noninvasive therapeutic treatments or diagnostics purpose. The study of plasma holds promise for a myriad of applications ranging from lasers and electronics, hazardous decontamination, sterilization and disinfection of foods, soil, water, instruments, to medical uses in wound healing and treating certain types of tumors and cancers. Plasma represents a new state-of-the-art sterilization and disinfection treatment for certain oral and environmental pathogens, heat-sensitive materials, hard and soft surfaces, and may assist health care facilities in the management of various health concerns. The role that low temperature atmospheric pressure plasma (LTAPP could play in the inactivation of pathogenic microorganisms might prove to be a new, faster, more economical alternative.

  5. Arc Plasma Torch Modeling

    CERN Document Server

    Trelles, J P; Vardelle, A; Heberlein, J V R

    2013-01-01

    Arc plasma torches are the primary components of various industrial thermal plasma processes involving plasma spraying, metal cutting and welding, thermal plasma CVD, metal melting and remelting, waste treatment and gas production. They are relatively simple devices whose operation implies intricate thermal, chemical, electrical, and fluid dynamics phenomena. Modeling may be used as a means to better understand the physical processes involved in their operation. This paper presents an overview of the main aspects involved in the modeling of DC arc plasma torches: the mathematical models including thermodynamic and chemical non-equilibrium models, turbulent and radiative transport, thermodynamic and transport property calculation, boundary conditions and arc reattachment models. It focuses on the conventional plasma torches used for plasma spraying that include a hot-cathode and a nozzle anode.

  6. Laminar Plasma Dynamos

    CERN Document Server

    Wang, Z; Barnes, C W; Barnes, D C; Wang, Zhehui; Pariev, Vladimir I.; Barnes, Cris W.; Barnes, Daniel C.

    2002-01-01

    A new kind of dynamo utilizing flowing laboratory plasmas has been identified. Conversion of plasma kinetic energy to magnetic energy is verified numerically by kinematic dynamo simulations for magnetic Reynolds numbers above 210. As opposed to intrinsically-turbulent liquid-sodium dynamos, the proposed plasma dynamos correspond to laminar flow topology. Modest plasma parameters, 1-20 eV temperatures, 10^{19}-10^{20} m^{-3} densities in 0.3-1.0 m scale-lengths driven by velocities on the order of the Alfven Critical Ionization Velocity (CIV), self-consistently satisfy the conditions needed for the magnetic field amplication. Growth rates for the plasma dynamos are obtained numerically with different geometry and magnetic Reynolds numbers. Magnetic-field-free coaxial plasma guns can be used to sustain the plasma flow and the dynamo.

  7. Plasma polymerization by Softplasma

    DEFF Research Database (Denmark)

    Jiang, J.; Wu, Zhenning; Benter, Maike

    2008-01-01

    In the late 19th century, the first depositions - known today as plasma polymers, were reported. In the last century, more and more research has been put into plasma polymers. Many different deposition systems have been developed. [1, 2] Shi F. F. broadly classified them into internal electrode......, external electrode, and electrodeless microwave or high frequency reactors. [3] Softplasma™ is an internal electrode plasma setup powered by low frequenc~ gower supply. It was developed in late 90s for surface treatment of silicone rubber. [ ]- 5] It is a low pressure, low electron density, 3D homogenous...... plasma. In this study, we are presenting the surface modification"pf polymers by plasma polymerization using Softplasma™. Softplasma™ can be used for two major types of polymerization: polymerization of vinyl monomers, where plasma acts as initiator; chemical vapour deposition, where plasma acts...

  8. Plasmas for medicine

    Science.gov (United States)

    von Woedtke, Th.; Reuter, S.; Masur, K.; Weltmann, K.-D.

    2013-09-01

    Plasma medicine is an innovative and emerging field combining plasma physics, life science and clinical medicine. In a more general perspective, medical application of physical plasma can be subdivided into two principal approaches. (i) “Indirect” use of plasma-based or plasma-supplemented techniques to treat surfaces, materials or devices to realize specific qualities for subsequent special medical applications, and (ii) application of physical plasma on or in the human (or animal) body to realize therapeutic effects based on direct interaction of plasma with living tissue. The field of plasma applications for the treatment of medical materials or devices is intensively researched and partially well established for several years. However, plasma medicine in the sense of its actual definition as a new field of research focuses on the use of plasma technology in the treatment of living cells, tissues, and organs. Therefore, the aim of the new research field of plasma medicine is the exploitation of a much more differentiated interaction of specific plasma components with specific structural as well as functional elements or functionalities of living cells. This interaction can possibly lead either to stimulation or inhibition of cellular function and be finally used for therapeutic purposes. During recent years a broad spectrum of different plasma sources with various names dedicated for biomedical applications has been reported. So far, research activities were mainly focused on barrier discharges and plasma jets working at atmospheric pressure. Most efforts to realize plasma application directly on or in the human (or animal) body for medical purposes is concentrated on the broad field of dermatology including wound healing, but also includes cancer treatment, endoscopy, or dentistry. Despite the fact that the field of plasma medicine is very young and until now mostly in an empirical stage of development yet, there are first indicators of its enormous

  9. Nonlinear Plasma Wave in Magnetized Plasmas

    CERN Document Server

    Bulanov, Sergei V; Kando, Masaki; Koga, James K; Hosokai, Tomonao; Zhidkov, Alexei G; Kodama, Ryosuke

    2013-01-01

    Nonlinear axisymmetric cylindrical plasma oscillations in magnetized collisionless plasmas are a model for the electron fluid collapse on the axis behind an ultrashort relativisically intense laser pulse exciting a plasma wake wave. We present an analytical description of the strongly nonlinear oscillations showing that the magnetic field prevents closing of the cavity formed behind the laser pulse. This effect is demonstrated with 3D PIC simulations of the laser-plasma interaction. An analysis of the betatron oscillations of fast electrons in the presence of the magnetic field reveals a characteristic "Four-Ray Star" pattern which has been observed in the image of the electron bunch in experiments [T. Hosokai, et al., Phys. Rev. Lett. 97, 075004 (2006)].

  10. Identification of consensus binding sites clarifies FMRP binding determinants.

    Science.gov (United States)

    Anderson, Bart R; Chopra, Pankaj; Suhl, Joshua A; Warren, Stephen T; Bassell, Gary J

    2016-08-19

    Fragile X mental retardation protein (FMRP) is a multifunctional RNA-binding protein with crucial roles in neuronal development and function. Efforts aimed at elucidating how FMRP target mRNAs are selected have produced divergent sets of target mRNA and putative FMRP-bound motifs, and a clear understanding of FMRP's binding determinants has been lacking. To clarify FMRP's binding to its target mRNAs, we produced a shared dataset of FMRP consensus binding sequences (FCBS), which were reproducibly identified in two published FMRP CLIP sequencing datasets. This comparative dataset revealed that of the various sequence and structural motifs that have been proposed to specify FMRP binding, the short sequence motifs TGGA and GAC were corroborated, and a novel TAY motif was identified. In addition, the distribution of the FCBS set demonstrates that FMRP preferentially binds to the coding region of its targets but also revealed binding along 3' UTRs in a subset of target mRNAs. Beyond probing these putative motifs, the FCBS dataset of reproducibly identified FMRP binding sites is a valuable tool for investigating FMRP targets and function. © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

  11. Determination of activated plasma fibronectin using radioactive labelled collagen I

    DEFF Research Database (Denmark)

    Fenger, M

    1984-01-01

    The plasma concentration of biological active fibronectin was assayed by a protein binding assay using 125I-collagen I as ligand and heparin as activator. The standard curve is linear for a fibronectin range of 1.1-11 pmol (0.5-5.0 micrograms) and the coefficient of variation was less than 10%. T...

  12. Determination of activated plasma fibronectin using radioactive labelled collagen I

    DEFF Research Database (Denmark)

    Fenger, M

    1984-01-01

    The plasma concentration of biological active fibronectin was assayed by a protein binding assay using 125I-collagen I as ligand and heparin as activator. The standard curve is linear for a fibronectin range of 1.1-11 pmol (0.5-5.0 micrograms) and the coefficient of variation was less than 10...

  13. Antioxidant flavonoids bind human serum albumin

    Science.gov (United States)

    Kanakis, C. D.; Tarantilis, P. A.; Polissiou, M. G.; Diamantoglou, S.; Tajmir-Riahi, H. A.

    2006-10-01

    Human serum albumin (HSA) is a principal extracellular protein with a high concentration in blood plasma and carrier for many drugs to different molecular targets. Flavonoids are powerful antioxidants and prevent DNA damage. The antioxidative protections are related to their binding modes to DNA duplex and complexation with free radicals in vivo. However, flavonoids are known to inhibit the activities of several enzymes such as calcium phospholipid-dependent protein kinase, tyrosine protein kinase from rat lung, phosphorylase kinase, phosphatidylinositol 3-kinase and DNA topoisomerases that exhibit the importance of flavonoid-protein interaction. This study was designed to examine the interaction of human serum albumin (HSA) with quercetin (que), kaempferol (kae) and delphinidin (del) in aqueous solution at physiological conditions, using constant protein concentration of 0.25 mM (final) and various drug contents of 1 μM-1 mM. FTIR and UV-vis spectroscopic methods were used to determine the polyphenolic binding mode, the binding constant and the effects of flavonoid complexation on protein secondary structure. The spectroscopic results showed that flavonoids are located along the polypeptide chains through H-bonding interactions with overall affinity constant of Kque = 1.4 × 10 4 M -1, Kkae = 2.6 × 10 5 M -1 and Kdel = 4.71 × 10 5 M -1. The protein secondary structure showed no alterations at low pigment concentration (1 μM), whereas at high flavonoid content (1 mM), major reduction of α-helix from 55% (free HSA) to 42-46% and increase of β-sheet from 15% (free HSA) to 17-19% and β-anti from 7% (free HSA) to 10-20% occurred in the flavonoid-HSA adducts. The major reduction of HSA α-helix is indicative of a partial protein unfolding upon flavonoid interaction.

  14. Relationship between Plasma Albumin Concentration and Plasma Volume in 5 Inbred Rat Strains.

    Science.gov (United States)

    Rose, Rajiv; Klemcke, Harold G

    2015-09-01

    Using the Evans Blue procedure, we previously found strain-related differences in plasma volumes in 5 inbred rat strains. Because albumin binds strongly with Evans blue, this protein is important in the Evans blue method of plasma volume determination. Therefore, we speculated that interstrain differences in plasma albumin concentration (PAC) could distort calculated plasma volumes. To address this concern, we used ELISA techniques to measure PAC in these inbred rat strains. In study A, the blood volume was measured by using Evans blue dye, and albumin was measured at the start of hemorrhage. In study B, blood volume was not measured, and albumin was measured twice, near the start and end of hemorrhage (approximately 14 min apart). Neither study revealed any interstrain differences in PAC, which decreased after hemorrhage in all 5 strains. No correlation was found between PAC and plasma volume, survival time, blood lactate, or blood base excess. Percentage changes in PAC during hemorrhage were greater in salt-sensitive compared with Lewis rats. Moreover, these percentage changes were associated with survival time in Fawn hooded hypertensive rats. Our data show that the plasma volumes we measured previously were not misrepresented due to variations in PAC.

  15. Signal transduction by erythrocytes on specific binding of doxorubicin immobilized on nanodispersed magnetite

    Energy Technology Data Exchange (ETDEWEB)

    Mykhaylyk, Olga [Institute Applied Problems Physics and Biophysics, NAS, Sluzhbova 3, UA-03142 Kyiv (Ukraine)]. E-mail: Olga.Mykhaylyk@gmx.net; Kotzuruba, Anatoliy [Institute of Biochemistry, NAS, Leontovicha 9, UA-01030 Kyiv (Ukraine); Dudchenko, Nataliya [Institute Applied Problems Physics and Biophysics, NAS, Sluzhbova 3, UA-03142 Kyiv (Ukraine); Toerok, Gyula [Research Institute for Solid State Physics and Optics, H-1525 Budapest, P.O. Box 49 (Hungary)

    2005-05-15

    Two specific binding sites for doxorubicin were revealed at the plasma membrane of human erythrocytes on investigation of the binding of doxorubicin magnetic nanoconjugates. Free and conjugated doxorubicins modulated signal transduction in erythrocytes in a similar way. Both up-regulated nitric oxide and cyclic GMP (cGMP) and down-regulated cyclic AMP (cAMP) production and stabilize the membranes of damaged erythrocytes.

  16. Plasma Biomedicine in Orthopedics

    Science.gov (United States)

    Hamaguchi, Satsohi

    2012-10-01

    Various effects of plasmas irradiation on cells, tissues, and biomaterials relevant for orthopedic applications have been examined. For direct application of plasmas to living cells or tissues, dielectric barrier discharges (DBDs) with helium flows into ambient air were used. For biomaterial processing, on the other hand, either helium DBDs mentioned above or low-pressure discharges generated in a chamber were used. In this presentation, plasma effects on cell proliferation and plasma treatment for artificial bones will be discussed. First, the conditions for enhanced cell proliferation in vitro by plasma applications have been examined. The discharge conditions for cell proliferation depend sensitively on cell types. Since cell proliferation can be enhanced even when the cells are cultured in a plasma pre-treated medium, long-life reactive species generated in the medium by plasma application or large molecules (such as proteins) in the medium modified by the plasma are likely to be the cause of cell proliferation. It has been found that there is strong correlation between (organic) hydroperoxide generation and cell proliferation. Second, effects of plasma-treated artificial bones made of porous hydroxyapatite (HA) have been examined in vitro and vivo. It has been found that plasma treatment increases hydrophilicity of the surfaces of microscopic inner pores, which directly or indirectly promotes differentiation of mesenchymal stem cells introduced into the pores and therefore causes faster bone growth. The work has been performed in collaboration with Prof. H. Yoshikawa and his group members at the School of Medicine, Osaka University.

  17. Plasma detachment in linear devices

    Science.gov (United States)

    Ohno, N.

    2017-03-01

    Plasma detachment research in linear devices, sometimes called divertor plasma simulators, is reviewed. Pioneering works exploring the concept of plasma detachment were conducted in linear devices. Linear devices have contributed greatly to the basic understanding of plasma detachment such as volume plasma recombination processes, detached plasma structure associated with particle and energy transport, and other related issues including enhancement of convective plasma transport, dynamic response of plasma detachment, plasma flow reversal, and magnetic field effect. The importance of plasma detachment research using linear devices will be highlighted aimed at the design of future DEMO.

  18. Advanced plasma diagnostics for plasma processing

    Science.gov (United States)

    Malyshev, Mikhail Victorovich

    1999-10-01

    A new, non-intrusive, non-perturbing diagnostic method was developed that can be broadly applied to low pressure, weakly ionized plasmas and glow discharges-trace rare gases optical emission spectroscopy (TRG-OES). The method is based on a comparison of intensities of atomic emission from trace amounts of inert gases (He, Ne, Ar, Kr, and Xe) that are added to the discharge to intensities calculated from the theoretical model. The model assumes a Maxwellian electron energy distribution function (EEDF), computes the population of emitting levels both from the ground state and the metastable states of rare gases, and from the best fit between theory and experiment determines electron temperature (Te). Subject to conditions, TRG-OES can also yield electron density or its upper or lower limit. From the comparison of the emission from levels excited predominantly by high energy electrons to that excited by low energy electrons, information about the EEDF can be obtained. The use of TRG-OES also allows a traditionally qualitative actinometry technique (determination of concentration of radical species in plasma through optical emission) to become a precise quantitative method by including Te and rare gases metastables effects. A combination of TRG-OES, advanced actinometry, and Langmuir probe measurements was applied to several different plasma reactors and regimes of operation. Te measurements and experiments to correct excitation cross section were conducted in a laboratory helical resonator. Two chamber configuration of a commercial (Lam Research) metal etcher were studied to determine the effects of plasma parameters on plasma-induced damage. Two different methods (RF inductive coupling and ultra-high frequency coupling) for generating a plasma in a prototype reactor were also studied. Pulsed plasmas, a potential candidate to eliminate the plasma-induced damage to microelectronics devices that occurs in manufacturing due to differential charging of the wafer, have

  19. Binding Energy and Enzymatic Catalysis.

    Science.gov (United States)

    Hansen, David E.; Raines, Ronald T.

    1990-01-01

    Discussed is the fundamental role that the favorable free energy of binding of the rate-determining transition state plays in catalysis. The principle that all of the catalytic factors discussed are realized by the use of this binding energy is reviewed. (CW)

  20. The electron-atom interaction in partially ionized dense plasmas

    Energy Technology Data Exchange (ETDEWEB)

    Omarbakiyeva, Yu A; Ramazanov, T S; Roepke, G [IETP, Al Farabi Kazakh National University, Tole Bi 96a, Almaty 050012 (Kazakhstan)], E-mail: yultuz@physics.kz

    2009-05-29

    The electron-atom interaction is considered in dense partially ionized plasmas. The separable potential is constructed from scattering data using effective radius theory. Parameters of the interaction potential were obtained from phase shifts, scattering length and effective radius. The binding energy of the electron in the H{sup -} ion is determined for the singlet channel on the basis of the reconstructed separable potential. In dense plasmas, the influence of the Pauli exclusion principle on the phase shifts and the binding energy is considered. Due to the Pauli blocking, the binding energy vanishes at the Mott density. At that density the behavior of the phase shifts is drastically changed. This leads to modifications of macroscopic properties such as composition and transport coefficients.

  1. Expressions of effective molecules transcriptional coactivator with plasma dissociated zircon-binding motif of Hippo signaling pathway in human osteosarcomas and osteosarcoma stem cells%Hippo信号通路作用分子TAZ在人骨肉瘤及骨肉瘤干细胞中的表达

    Institute of Scientific and Technical Information of China (English)

    朱忠胜; 张春林; 汪泱

    2014-01-01

    目的:检测在人骨肉瘤组织、骨肉瘤MG63细胞及其干细胞中Hippo信号分子TAZ的表达情况,比较骨肉瘤细胞和骨肉瘤干细胞中 TAZ 的表达差异,并探讨其可能的临床意义。方法2010年1月至2012年1月在我院治疗的骨肉瘤患者中,选取原发骨肉瘤组织标本12例,复发的骨肉瘤标本6例,另选6例骨纤维结构不良组织作阴性对照,并贴壁培养骨肉瘤MG63细胞,采用免疫组化方法检测骨肉瘤组织和骨肉瘤MG63细胞中 TAZ 的表达情况;用无血清悬浮培养骨肉瘤 MG63细胞,分离并收集骨肉瘤细胞球,通过定量反转录-聚合酶链反应( RT-PCR )法检测骨肉瘤MG63细胞及骨肉瘤细胞球中胚胎干细胞标志基因( Nanog, Oct4)和 Hippo 信号分子 TAZ 的表达,Western 印记法检测 MG63细胞和细胞球中 TAZ 蛋白的表达情况。结果在12例人骨肉瘤原发组织标本中,3例骨肉瘤组织TAZ表达阳性,而6例骨肉瘤复发组织中TAZ表达全部阳性,在骨肉瘤细胞MG63中TAZ分子表达也呈阳性,而骨纤维结果不良中全部表达阴性。与骨肉瘤MG63细胞相比,RT-PCR显示培养的骨肉瘤细胞球中干细胞标志基因Nanog,Oct4和Hippo信号分子TAZ明显高表达。Western blot 提示细胞球中 TAZ 蛋白表达明显高于骨肉瘤 MG63细胞。结论骨肉瘤组织中存在TAZ分子的表达,在复发骨肉瘤组织中呈高表达;在骨肉瘤细胞株中也存在TAZ分子的表达,而在干细胞中TAZ分子表达更高;提示其与骨肉瘤干细胞的特征具有一定的相关性。%Objective To detect the expressions of transcriptional coactivator with plasma dissociated zircon ( PDZ )-binding motif ( TAZ ) molecules of Hippo signaling pathway in human osteosarcoma tissues, osteosarcoma MG63 cells and stem cells, to compare the expression differences of TAZ molecules in osteosarcoma cells and osteosarcoma stem cells, and to investigate possible clinical

  2. Prognostic values of heart-type fatty acid-binding protein and D-dimer level in plasma in acute pulmonary embolism patients%心肌型脂肪酸结合蛋白联合血浆D-二聚体对肺栓塞预后评估的临床意义

    Institute of Scientific and Technical Information of China (English)

    陶琳; 杨毅

    2015-01-01

    ObjectiveTo evaluate the clinical value of heart-type fatty acid-binding protein (H-FABP) and D-dimer in assessing the prognosis of acute pulmonary embolism (APE). MethodsTotaly 120 APE patients were hospitalized from January 2011 to December 2014 and enroled in this study. The plasma H-FABP and D-dimer level were measured by enzyme-linked immunosorbent assay. The APE patients were divided three groups including low-risk, middle-risk and high-risk group by European heart association new guidelines. According to the clinical prognosis, the APE patients were divided into survival and death group. The H-FABP and D-dimer level in different groups and the relationship with the prognosis were assessed.ResultsWith the increased severity in patients, the H-FABP and D-dimer level were significantly elevated (P<0.05); the H-FABP and D-dimer level were significantly higher in death group as compared with survival group (P<0.05). The H-FABP and D-dimer level were positively correlated (r=0.693,P=0.000). ROC curves analysis results showed that the area under curve of H-FABP was 0.845 (95%CI: 0.752-0.918), and optimal operating point (OOP) was 8.65 µg/L, which had 81.24% sensitivity and 84.14% specificity; ACU of D-dimer was 0.832 (95%CI: 0.728-0.899), and OOP was 1.25 mg/L, which had 83.72% sensitivity and 82.65% specificity.Conclusion The H-FABP and D-dimer can effectively assess severity and prognosis of APE patients, meanwhile, it provide an objective basis for the clinical individual treatment and reducing the mortality rate of APE patients.%目的:探讨心肌型脂肪酸结合蛋白(H-FABP )联合血浆 D-二聚体在评价急性肺栓塞(APE)患者预后中的临床价值。方法选取本院2011年1月至2014年12月确诊的APE患者120例为研究对象,采用酶联免疫吸附法分别测定外周血H-FABP及D-二聚体浓度。根据病情严重程度将APE患者分为低危组、中危组及高危组;根据临床转归,分为存活组和死亡组

  3. 人视黄醇结合蛋白4水平升高与冠心病及其预后相关%Elevated plasma levels of retinol-binding protein-4 are associated coronary artery disease and one-year prognosis

    Institute of Scientific and Technical Information of China (English)

    谢芳艺; 陈忠; 丁震; 范沛英; 王鑫; 冯毅; 马根山

    2012-01-01

    Objective: To examine whether plasma levels of retinol-binding protein-4 (RBP4) are associated with coronary artery disease (CAD) and one-year prognosis. Methods: RBP4 levels were determined by ELISA kit (IBL. Germany) in 140 patients without type 2 diabetes undergoing coronary angiography for the concern of CAD. Patients were divided into controls (normal coronary arteries, n =46) and CAD (≥50% diameter stenosis, n = 94). CAD patients were followed up for average 12 months. Major adverse cardiac events (MACEs) including recurrent anginal, recurrent infarction, deteriorated heart failure and cardiac death were analyzed. Multivariate analysis was done to determine the association between RBP4 and composite MACEs during one- year follow up. Results: Compared to the controls, patients with CAD were older, had higher prevalence of hypertension and smokers (all P <0. 05). Meanwhile, patients suffering from MACEs (n = 8) had higher levels of RBP4 than those without [ (76. 12 ± 42. 89) μg·ml-1 vs (46. 24 ± 17. 66) μg·ml-1 , P <0. 01 ] ; these patients suffering from MACEs had higher ratio of RBP4 levels 3=70 μg · ml-1 than those without (37.50% vs 9.30%, P =0.018). Multivariate analysis demonstrated that RBP4 was an independent predictor of composite MACEs during one-year follow up ( OR: 1. 091 , 95% CI:1.015-1. 172, P =0. 018) . Conclusion-. RBP-4 levels are associated with CAD and one-year prognosis. Patients with higher RBP-4 levels are at higher risk of MACEs.%目的:研究人视黄醇结合蛋白4 (RBP4)水平与冠心病及1年预后关系.方法:选择因疑诊冠心病而行冠状动脉造影的140例非糖尿病患者,其中确诊冠心病患者94例,非冠心病患者46例(作为对照).采用ELISA试剂盒(德国IBL公司)检测140例患者血浆RBP4水平.冠心病患者出院后完成1年随访,记录主要不良心脏事件(MACEs),即再发心绞痛、再发心肌梗死、心力衰竭恶化和心源性死亡.多变量回归分析RBP4与1年随

  4. Pulsed plasma arc cladding

    Institute of Scientific and Technical Information of China (English)

    龙; 白钢; 李振民; 张赋升; 杨思乾

    2004-01-01

    A prototype of Pulsed Plasma Arc Cladding system was developed, in which single power source supplies both transferred plasma arc (TPA) and non-transferred plasma arc (N-TPA). Both plasmas work in turn in a high frequency controlled by an IGBT connecting nozzle and workpiece. The working frequency of IGBT ranges from 50 ~ 7000Hz, in which the plasmas can work in turn smoothly. Higher than 500 Hz of working frequency is suggested for promotion of cladding quality and protection of IGBT. Drag phenomenon of TPA intensifies as the frequency goes up, which tends to increase the current proportion of TPA and suppress N-TPA. The occupation ratio of IGBT can be regulated from 5% ~ 95%, which balances the power supplies of both plasmas. An occupation ratio higher than 50% gives adequate proportion of arc current for N-TPA to preheat powder.

  5. Introduction to Complex Plasmas

    CERN Document Server

    Bonitz, Michael; Ludwig, Patrick

    2010-01-01

    Complex plasmas differ from traditional plasmas in many ways: these are low-temperature high pressure systems containing nanometer to micrometer size particles which may be highly charged and strongly interacting. The particles may be chemically reacting or be in contact with solid surfaces, and the electrons may show quantum behaviour. These interesting properties have led to many applications of complex plasmas in technology, medicine and science. Yet complex plasmas are extremely complicated, both experimentally and theoretically, and require a variety of new approaches which go beyond standard plasma physics courses. This book fills this gap presenting an introduction to theory, experiment and computer simulation in this field. Based on tutorial lectures at a very successful recent Summer Institute, the presentation is ideally suited for graduate students, plasma physicists and experienced undergraduates.

  6. Ultracold Neutral Plasmas

    CERN Document Server

    Killian, T C; Gupta, P; Laha, S; Martinez, Y N; Mickelson, P G; Nagel, S B; Saenz, A D; Simien, C E; Killian, Thomas C.

    2005-01-01

    Ultracold neutral plasmas are formed by photoionizing laser-cooled atoms near the ionization threshold. Through the application of atomic physics techniques and diagnostics, these experiments stretch the boundaries of traditional neutral plasma physics. The electron temperature in these plasmas ranges from 1-1000 K and the ion temperature is around 1 K. The density can approach $10^{11}$ cm$^{-3}$. Fundamental interest stems from the possibility of creating strongly-coupled plasmas, but recombination, collective modes, and thermalization in these systems have also been studied. Optical absorption images of a strontium plasma, using the Sr$^+$ ${^2S_{1/2}} -> {^2P_{1/2}}$ transition at 422 nm, depict the density profile of the plasma, and probe kinetics on a 50 ns time-scale. The Doppler-broadened ion absorption spectrum measures the ion velocity distribution, which gives an accurate measure of the ion dynamics in the first microsecond after photoionization.

  7. What is a plasma?

    Energy Technology Data Exchange (ETDEWEB)

    Intrator, Thomas P. [Los Alamos National Laboratory

    2012-08-30

    This introduction will define the plasma fourth state of matter, where we find plasmas on earth and beyond, and why they are useful. There are applications to many consumer items, fusion energy, scientific devices, satellite communications, semiconductor processing, spacecraft propulsion, and more. Since 99% of our observable universe is ionized gas, plasma physics determines many important features of astrophysics, space physics, and magnetosphere physics in our solar system. We describe some plasma characteristics, examples in nature, some useful applications, how to create plasmas. A brief introduction to the theoretical framework includes the connection between kinetic and fluid descriptions, quasi neutrality, Debye shielding, ambipolar electric fields, some plasma waves. Hands-on demonstrations follow. More complete explanations will follow next week.

  8. Pulsed plasma electron sourcesa)

    Science.gov (United States)

    Krasik, Ya. E.; Yarmolich, D.; Gleizer, J. Z.; Vekselman, V.; Hadas, Y.; Gurovich, V. Tz.; Felsteiner, J.

    2009-05-01

    There is a continuous interest in research of electron sources which can be used for generation of uniform electron beams produced at E ≤105 V/cm and duration ≤10-5 s. In this review, several types of plasma electron sources will be considered, namely, passive (metal ceramic, velvet and carbon fiber with and without CsI coating, and multicapillary and multislot cathodes) and active (ferroelectric and hollow anodes) plasma sources. The operation of passive sources is governed by the formation of flashover plasma whose parameters depend on the amplitude and rise time of the accelerating electric field. In the case of ferroelectric and hollow-anode plasma sources the plasma parameters are controlled by the driving pulse and discharge current, respectively. Using different time- and space-resolved electrical, optical, spectroscopical, Thomson scattering and x-ray diagnostics, the parameters of the plasma and generated electron beam were characterized.

  9. Binding of straight-chain saturated dicarboxylic acids to albumin.

    Science.gov (United States)

    Tonsgard, J H; Mendelson, S A; Meredith, S C

    1988-11-01

    Dicarboxylic acids are prominent features of several diseases, including Reye's syndrome. Long-chain dicarboxylic acids have profound effects on the function and structure of isolated mitochondria, suggesting that they could contribute to the mitochondrial dysfunction in Reye's syndrome. Binding of fatty acids to albumin and the intracellular fatty acid-binding proteins is important in regulating the transport and metabolism of fatty acids and protects against the toxic effects of unbound fatty acids. We studied the binding of dicarboxylic acids to defatted albumin using equilibrium dialysis to assess to what extent dicarboxylic acids are likely to be bound in the plasma of patients. Dicarboxylic acids bind weakly to albumin in a molar ratio of 3.8, 4.2, 1.6, 0.8, and 0.7 to 1 for octadecanedioic, hexadecanedioic, tetradecanedioic, dodecanedioic, and decanedioic acid, respectively. The dissociation constants for long-chain dicarboxylic acids are 100-1,000-fold larger than those of comparable monocarboxylic acids. Oleate competes with dicarboxylic acid and reduces the moles of dicarboxylic acid bound per mol of albumin to less than 1. Octanoate inhibits dicarboxylic acid binding. Our observations indicate that in Reye's syndrome, substantial concentrations of dicarboxylic acids of patients may be free and potentially toxic to mitochondria and other cellular processes.

  10. Radioiodination of chicken luteinizing hormone without affecting receptor binding potency

    Energy Technology Data Exchange (ETDEWEB)

    Kikuchi, M.; Ishii, S. (Waseda Univ., Tokyo (Japan))

    1989-12-01

    By improving the currently used lactoperoxidase method, we were able to obtain radioiodinated chicken luteinizing hormone (LH) that shows high specific binding and low nonspecific binding to a crude plasma membrane fraction of testicular cells of the domestic fowl and the Japanese quail, and to the ovarian granulosa cells of the Japanese quail. The change we made from the original method consisted of (1) using chicken LH for radioiodination that was not only highly purified but also retained a high receptor binding potency; (2) controlling the level of incorporation of radioiodine into chicken LH molecules by employing a short reaction time and low temperature; and (3) fractionating radioiodinated chicken LH further by gel filtration using high-performance liquid chromatography. Specific radioactivity of the final {sup 125}I-labeled chicken LH preparation was 14 microCi/micrograms. When specific binding was 12-16%, nonspecific binding was as low as 2-4% in the gonadal receptors. {sup 125}I-Labeled chicken LH was displaced by chicken LH and ovine LH but not by chicken follicle-stimulating hormone. The equilibrium association constant of quail testicular receptor was 3.6 x 10(9) M-1. We concluded that chicken LH radioiodinated by the present method is useful for studies of avian LH receptors.

  11. Space plasma physics research

    Science.gov (United States)

    Comfort, Richard H.; Horwitz, James L.

    1993-01-01

    During the course of this grant, work was performed on a variety of topics and there were a number of significant accomplishments. A summary of these accomplishments is included. The topics studied include empirical model data base, data reduction for archiving, semikinetic modeling of low energy plasma in the inner terrestrial magnetosphere and ionosphere, O(+) outflows, equatorial plasma trough, and plasma wave ray-tracing studies. A list of publications and presentations which have resulted from this research is also included.

  12. Atmospheric Plasma Depainting

    Science.gov (United States)

    2014-11-19

    Plasma Carbon Dioxide Water Vapor 11 Atmospheric Plasma Depainting, ASETSDefense, Nov 19, 2014 Features and Benefits of APCR Technology Feature...Depainting, ASETSDefense, Nov 19, 2014 14 APC on Aluminum Removal to Primer RAM on Carbon Fiber Partial Topcoat Removal APC Topcoat RAM...60Hz Plasma Flux™ Power Supply VENT To Facility HEPA <= Filtration COTS Six-Axis Robot Aircraft part Particulate Collection System

  13. Plasma nitriding of steels

    CERN Document Server

    Aghajani, Hossein

    2017-01-01

    This book focuses on the effect of plasma nitriding on the properties of steels. Parameters of different grades of steels are considered, such as structural and constructional steels, stainless steels and tools steels. The reader will find within the text an introduction to nitriding treatment, the basis of plasma and its roll in nitriding. The authors also address the advantages and disadvantages of plasma nitriding in comparison with other nitriding methods. .

  14. Plasma adiabatic lapse rate

    CERN Document Server

    Amendt, Peter; Wilks, Scott

    2012-01-01

    The plasma analog of an adiabatic lapse rate (or temperature variation with height) in atmospheric physics is obtained. A new source of plasma temperature gradient in a binary ion species mixture is found that is proportional to the concentration gradient and difference in average ionization states . Application to inertial-confinement-fusion implosions indicates a potentially strong effect in plastic (CH) ablators that is not modeled with mainline (single-fluid) simulations. An associated plasma thermodiffusion coefficient is derived, and charge-state diffusion in a single-species plasma is also predicted.

  15. Nonlinear Physics of Plasmas

    CERN Document Server

    Kono, Mitsuo

    2010-01-01

    A nonlinearity is one of the most important notions in modern physics. A plasma is rich in nonlinearities and provides a variety of behaviors inherent to instabilities, coherent wave structures and turbulence. The book covers the basic concepts and mathematical methods, necessary to comprehend nonlinear problems widely encountered in contemporary plasmas, but also in other fields of physics and current research on self-organized structures and magnetized plasma turbulence. The analyses make use of strongly nonlinear models solved by analytical techniques backed by extensive simulations and available experiments. The text is written for senior undergraduates, graduate students, lecturers and researchers in laboratory, space and fusion plasmas.

  16. Physics of Plasmas

    CERN Document Server

    Woods, Leslie Colin

    2003-01-01

    A short, self-sufficient introduction to the physics of plasma for beginners as well as researchers in a number of fields. The author looks at the dynamics and stability of magnetoplasma and discusses wave and transport in this medium. He also looks at such applications as fusion research using magnetic confinement of Deuterium plasma, solar physics with its plasma loops reaching high into the corona, sunspots and solar wind, engineering applications to metallurgy, MHD direct generation of electricity, and railguns, finally touching on the relatively new and difficult subject of dusty plasmas.

  17. Princeton Plasma Physics Laboratory

    Energy Technology Data Exchange (ETDEWEB)

    1990-01-01

    This report discusses the following topics: principal parameters achieved in experimental devices fiscal year 1990; tokamak fusion test reactor; compact ignition tokamak; Princeton beta experiment- modification; current drive experiment-upgrade; international collaboration; x-ray laser studies; spacecraft glow experiment; plasma processing: deposition and etching of thin films; theoretical studies; tokamak modeling; international thermonuclear experimental reactor; engineering department; project planning and safety office; quality assurance and reliability; technology transfer; administrative operations; PPPL patent invention disclosures for fiscal year 1990; graduate education; plasma physics; graduate education: plasma science and technology; science education program; and Princeton Plasma Physics Laboratory reports fiscal year 1990.

  18. Solid expellant plasma generator

    Science.gov (United States)

    Stone, Nobie H. (Inventor); Poe, Garrett D. (Inventor); Rood, Robert (Inventor)

    2010-01-01

    An improved solid expellant plasma generator has been developed. The plasma generator includes a support housing, an electrode rod located in the central portion of the housing, and a mass of solid expellant material that surrounds the electrode rod within the support housing. The electrode rod and the solid expellant material are made of separate materials that are selected so that the electrode and the solid expellant material decompose at the same rate when the plasma generator is ignited. This maintains a point of discharge of the plasma at the interface between the electrode and the solid expellant material.

  19. Plasma processing for VLSI

    CERN Document Server

    Einspruch, Norman G

    1984-01-01

    VLSI Electronics: Microstructure Science, Volume 8: Plasma Processing for VLSI (Very Large Scale Integration) discusses the utilization of plasmas for general semiconductor processing. It also includes expositions on advanced deposition of materials for metallization, lithographic methods that use plasmas as exposure sources and for multiple resist patterning, and device structures made possible by anisotropic etching.This volume is divided into four sections. It begins with the history of plasma processing, a discussion of some of the early developments and trends for VLSI. The second section

  20. Plasma and particles

    Science.gov (United States)

    Špatenka, Petr; Vacková, Tat'ana; Nováček, Vojtěch; Jeníková, Zdenka

    2016-12-01

    Plasma has been proved as a standard industrial method for surface treatment of solid bulk materials. Recently plasma has also been used in connection with production, treatment and functionalization of powder and granulate materials. Functionalization was originally developed for hydrophylization of hydrophobic surfaces of particles made from various materials. An industrial scale device with a capacity of several hundreds of tons per year based on plasma treatment will be presented. As examples of the applications are given plasma treated polyethylene powder dispersed in the water; and very good adhesion of polymer powders to metals or glass, which is promising for development of new generation of thermoplastic composites.

  1. ApoD mediates binding of HDL to LDL and to growing T24 carcinoma.

    Directory of Open Access Journals (Sweden)

    Sten Braesch-Andersen

    Full Text Available Apolipoprotein (Apo D is an important protein produced in many parts of the body. It is necessary for the development and repair of the brain and protection from oxidative stress. The purpose of this study was to investigate the extent to which apoD interacts with lipoproteins in human plasma. By using detergent-free ELISA, we show that immobilized monoclonal antibodies against apoD very efficiently bind to low density lipoprotein (LDL from plasma; this binding is as equally efficient as binding to an anti-apoB monoclonal antibody. Adding detergent to the plasma inhibited the binding, suggesting that the binding is dependent on the presence of intact lipoprotein particles. Reversing the system by using immobilized anti-apoB revealed that the affinity of apoD for LDL is rather low, suggesting that multiple bindings are needed for a durable connection. Biosensor experiments using purified lipoproteins also showed that purified apoD and high density lipoprotein 3 (HDL3, a lipoprotein fraction rich in apoD, were both able to bind LDL very efficiently, indicating that the HDL3-LDL interaction may be a physiological consequence of the affinity of apoD for LDL. Furthermore, we found that apoD increases the binding of HDL to actively growing T24 bladder carcinoma cells but not to quiescent, contact-inhibited, confluent T24 cells. This result is especially intriguing given that the T24 supernatant only contained detectable levels of apoD after growth inhibition, raising the possibility that alternating the expression of apoD and a putative apoD-receptor could give direction to the flow of lipids. In the current paper, we conclude that apoD mediates binding of HDL to LDL and to growing T24 carcinomas, thereby highlighting the importance of apoD in lipid metabolism.

  2. ApoD mediates binding of HDL to LDL and to growing T24 carcinoma.

    Science.gov (United States)

    Braesch-Andersen, Sten; Beckman, Lena; Paulie, Staffan; Kumagai-Braesch, Makiko

    2014-01-01

    Apolipoprotein (Apo) D is an important protein produced in many parts of the body. It is necessary for the development and repair of the brain and protection from oxidative stress. The purpose of this study was to investigate the extent to which apoD interacts with lipoproteins in human plasma. By using detergent-free ELISA, we show that immobilized monoclonal antibodies against apoD very efficiently bind to low density lipoprotein (LDL) from plasma; this binding is as equally efficient as binding to an anti-apoB monoclonal antibody. Adding detergent to the plasma inhibited the binding, suggesting that the binding is dependent on the presence of intact lipoprotein particles. Reversing the system by using immobilized anti-apoB revealed that the affinity of apoD for LDL is rather low, suggesting that multiple bindings are needed for a durable connection. Biosensor experiments using purified lipoproteins also showed that purified apoD and high density lipoprotein 3 (HDL3), a lipoprotein fraction rich in apoD, were both able to bind LDL very efficiently, indicating that the HDL3-LDL interaction may be a physiological consequence of the affinity of apoD for LDL. Furthermore, we found that apoD increases the binding of HDL to actively growing T24 bladder carcinoma cells but not to quiescent, contact-inhibited, confluent T24 cells. This result is especially intriguing given that the T24 supernatant only contained detectable levels of apoD after growth inhibition, raising the possibility that alternating the expression of apoD and a putative apoD-receptor could give direction to the flow of lipids. In the current paper, we conclude that apoD mediates binding of HDL to LDL and to growing T24 carcinomas, thereby highlighting the importance of apoD in lipid metabolism.

  3. Identification of fibrin clot-bound plasma proteins.

    Directory of Open Access Journals (Sweden)

    Simone Talens

    Full Text Available Several proteins are known to bind to a fibrin network and to change clot properties or function. In this study we aimed to get an overview of fibrin clot-bound plasma proteins. A plasma clot was formed by adding thrombin, CaCl(2 and aprotinin to citrated platelet-poor plasma and unbound proteins were washed away with Tris-buffered saline. Non-covalently bound proteins were extracted, separated with 2D gel electrophoresis and visualized with Sypro Ruby. Excised protein spots were analyzed with mass spectrometry. The identity of the proteins was verified by checking the mass of the protein, and, if necessary, by Western blot analysis. Next to established fibrin-binding proteins we identified several novel fibrin clot-bound plasma proteins, including α(2-macroglobulin, carboxypeptidase N, α(1-antitrypsin, haptoglobin, serum amyloid P, and the apolipoproteins A-I, E, J, and A-IV. The latter six proteins are associated with high-density lipoprotein particles. In addition we showed that high-density lipoprotein associated proteins were also present in fibrinogen preparations purified from plasma. Most plasma proteins in a fibrin clot can be classified into three groups according to either blood coagulation, protease inhibition or high-density lipoprotein metabolism. The presence of high-density lipoprotein in clots might point to a role in hemostasis.

  4. Cooperative binding: a multiple personality.

    Science.gov (United States)

    Martini, Johannes W R; Diambra, Luis; Habeck, Michael

    2016-06-01

    Cooperative binding has been described in many publications and has been related to or defined by several different properties of the binding behavior of the ligand to the target molecule. In addition to the commonly used Hill coefficient, other characteristics such as a sigmoidal shape of the overall titration curve in a linear plot, a change of ligand affinity of the other binding sites when a site of the target molecule becomes occupied, or complex roots of the binding polynomial have been used to define or to quantify cooperative binding. In this work, we analyze how the different properties are related in the most general model for binding curves based on the grand canonical partition function and present several examples which highlight differences between the cooperativity characterizing properties which are discussed. Our results mainly show that among the presented definitions there are not two which fully coincide. Moreover, this work poses the question whether it can make sense to distinguish between positive and negative cooperativity based on the macroscopic binding isotherm only. This article shall emphasize that scientists who investigate cooperative effects in biological systems could help avoiding misunderstandings by stating clearly which kind of cooperativity they discuss.

  5. Drug binding to human serum albumin: abridged review of results obtained with high-performance liquid chromatography and circular dichroism.

    Science.gov (United States)

    Ascoli, Giorgio A; Domenici, Enrico; Bertucci, Carlo

    2006-09-01

    The drug binding to plasma and tissue proteins are fundamental factors in determining the overall pharmacological activity of a drug. Human serum albumin (HSA), together with alpha1-acid glycoprotein (AGP), are the most important plasma proteins, which act as drug carriers, with drug pharmacokinetic implications, resulting in important clinical impacts for drugs that have a relatively narrow therapeutic index. This review focuses on the combination of biochromatography and circular dichroism as an effective approach for the characterization of albumin binding sites and their enantioselectivity. Furthermore, their applications to the study of changes in the binding properties of the protein arising by the reversible or covalent binding of drugs are discussed, and examples of physiological relevance reported. Perspectives of these studies reside in supporting the development of new drugs, which require miniaturization to facilitate the screening of classes of compounds for their binding to the target protein, and a deeper characterization of the mechanisms involved in the molecular recognition processes.

  6. Determination of human serum alpha1-acid glycoprotein and albumin binding of various marketed and preclinical kinase inhibitors.

    Science.gov (United States)

    Zsila, Ferenc; Fitos, Ilona; Bencze, Gyula; Kéri, György; Orfi, László

    2009-01-01

    There are about 380 protein kinase inhibitors in drug development as of today and 15 drugs have been marketed already for the treatment of cancer. This time 139 validated kinase targets are in the focus of drug research of pharmaceutical companies and big efforts are made for the development of new, druglike kinase inhibitors. Plasma protein binding is an important factor of the ADME profiling of a drug compound. Human serum albumin (HSA) and alpha(1)-acid glycoprotein (AAG) are the most relevant drug carriers in blood plasma. Since previous literature data indicated that AAG is the principal plasma binding component of some kinase inhibitors the present work focuses on the comprehensive evaluation of AAG binding of a series of marketed and experimental kinase inhibitors by using circular dichroism (CD) spectroscopy approach. HSA binding was also evaluated by affinity chromatography. Protein binding interactions of twenty-six kinase inhibitors are characterized. The contribution of AAG and HSA binding data to the pharmacokinetic profiles of the investigated therapeutic agents is discussed. Structural, biological and drug binding properties of AAG as well as the applicability of the CD method in studying drug-protein binding interactions are also briefly reviewed.

  7. Ultracold neutral plasmas

    Science.gov (United States)

    Lyon, M.; Rolston, S. L.

    2017-01-01

    By photoionizing samples of laser-cooled atoms with laser light tuned just above the ionization limit, plasmas can be created with electron and ion temperatures below 10 K. These ultracold neutral plasmas have extended the temperature bounds of plasma physics by two orders of magnitude. Table-top experiments, using many of the tools from atomic physics, allow for the study of plasma phenomena in this new regime with independent control over the density and temperature of the plasma through the excitation process. Characteristic of these systems is an inhomogeneous density profile, inherited from the density distribution of the laser-cooled neutral atom sample. Most work has dealt with unconfined plasmas in vacuum, which expand outward at velocities of order 100 m/s, governed by electron pressure, and with lifetimes of order 100 μs, limited by stray electric fields. Using detection of charged particles and optical detection techniques, a wide variety of properties and phenomena have been observed, including expansion dynamics, collective excitations in both the electrons and ions, and collisional properties. Through three-body recombination collisions, the plasmas rapidly form Rydberg atoms, and clouds of cold Rydberg atoms have been observed to spontaneously avalanche ionize to form plasmas. Of particular interest is the possibility of the formation of strongly coupled plasmas, where Coulomb forces dominate thermal motion and correlations become important. The strongest impediment to strong coupling is disorder-induced heating, a process in which Coulomb energy from an initially disordered sample is converted into thermal energy. This restricts electrons to a weakly coupled regime and leaves the ions barely within the strongly coupled regime. This review will give an overview of the field of ultracold neutral plasmas, from its inception in 1999 to current work, including efforts to increase strong coupling and effects on plasma properties due to strong coupling.

  8. Mechanism of Action and Clinical Application of Tafamidis in Hereditary Transthyretin Amyloidosis.

    Science.gov (United States)

    Coelho, Teresa; Merlini, Giampaolo; Bulawa, Christine E; Fleming, James A; Judge, Daniel P; Kelly, Jeffery W; Maurer, Mathew S; Planté-Bordeneuve, Violaine; Labaudinière, Richard; Mundayat, Rajiv; Riley, Steve; Lombardo, Ilise; Huertas, Pedro

    2016-06-01

    Transthyretin (TTR) transports the retinol-binding protein-vitamin A complex and is a minor transporter of thyroxine in blood. Its tetrameric structure undergoes rate-limiting dissociation and monomer misfolding, enabling TTR to aggregate or to become amyloidogenic. Mutations in the TTR gene generally destabilize the tetramer and/or accelerate tetramer dissociation, promoting amyloidogenesis. TTR-related amyloidoses are rare, fatal, protein-misfolding disorders, characterized by formation of soluble aggregates of variable structure and tissue deposition of amyloid. The TTR amyloidoses present with a spectrum of manifestations, encompassing progressive neuropathy and/or cardiomyopathy. Until recently, the only accepted treatment to halt progression of hereditary TTR amyloidosis was liver transplantation, which replaces the hepatic source of mutant TTR with the less amyloidogenic wild-type TTR. Tafamidis meglumine is a rationally designed, non-NSAID benzoxazole derivative that binds with high affinity and selectivity to TTR and kinetically stabilizes the tetramer, slowing monomer formation, misfolding, and amyloidogenesis. Tafamidis is the first pharmacotherapy approved to slow the progression of peripheral neurologic impairment in TTR familial amyloid polyneuropathy. Here we describe the mechanism of action of tafamidis and review the clinical data, demonstrating that tafamidis treatment slows neurologic deterioration and preserves nutritional status, as well as quality of life in patients with early-stage Val30Met amyloidosis.

  9. Plasma Physics An Introduction to Laboratory, Space, and Fusion Plasmas

    CERN Document Server

    Piel, Alexander

    2010-01-01

    Plasma Physics gives a comprehensive introduction to the basic processes in plasmas and demonstrates that the same fundamental concepts describe cold gas-discharge plasmas, space plasmas, and hot fusion plasmas. Starting from particle drifts in magnetic fields, the principles of magnetic confinement fusion are explained and compared with laser fusion. Collective processes are discussed in terms of plasma waves and instabilities. The concepts of plasma description by magnetohydrodynamics, kinetic theory, and particle simulation are stepwise introduced. Space charge effects in sheath regions, double layers and plasma diodes are given the necessary attention. The new fundamental mechanisms of dusty plasmas are explored and integrated into the framework of conventional plasmas. The book concludes with a brief introduction to plasma discharges. Written by an internationally renowned researcher in experimental plasma physics, the text keeps the mathematical apparatus simple and emphasizes the underlying concepts. T...

  10. Agglomeration of microparticles in complex plasmas

    CERN Document Server

    Du, Cheng-Ran; Ivlev, Alexei; Konopka, Uwe; Morfill, Gregor

    2010-01-01

    Agglomeration of highly charged microparticles was observed and studied in complex plasma experiments carried out in a capacitively coupled rf discharge. The agglomeration was caused by strong dust density waves triggered in a particle cloud by decreasing neutral gas pressure. Using a high-speed camera during this unstable regime, it was possible to resolve the motion of individual microparticles and to show that the relative velocities of some particles were sufficiently high to overcome the mutual Coulomb repulsion and hence to result in agglomeration. After stabilising the cloud again through the increase of the pressure, we were able to observe the aggregates directly with a long-distance microscope. We show that the agglomeration rate deduced from our experiments is in good agreement with theoretical estimates. In addition, we briefly discuss the mechanisms that can provide binding of highly charged microparticles in a plasma.

  11. Proteins of the canine seminal plasma

    Directory of Open Access Journals (Sweden)

    Annice Aquino-Cortez

    2016-05-01

    Full Text Available ABSTRACT: Studies have been performed to identify the proteins present in canine seminal plasma (SP and relate them to sperm quality as well as to discover molecular markers of reproductive tract diseases. There is evidence that heparin-binding proteins, zinc-binding proteins, and lactoferrin as well as the matrix metalloproteinase, superoxide dismutase, catalase, and glutathione peroxidase enzymes are associated with canine sperm quality. Other studies indicate that prolactin and enzymes like arginine esterase, acid phosphatase, and alkaline phosphatase could be successfully used as biomarkers of reproductive disorders. Thus, the present literature review aims to address aspects related to proteins of the canine SP, their influence on fertility, and their importance as biomarkers of reproductive disorders.

  12. Estimation of angiotensin-converting enzyme inhibitors protein binding degree using chromatographic hydrophobicity data

    Directory of Open Access Journals (Sweden)

    Trbojević-Stanković Jasna

    2015-01-01

    Full Text Available Introduction. Angiotensin-converting enzyme (ACE inhibitors represent a significant group of drugs primarily used in the treatment of hypertension and congestive heart failure. Objective. Selected ACE inhibitors (enalapril, quinapril, fosinopril, lisinopril, cilazapril were studied in order to establish a fast and easy estimation method of their plasma protein binding degree based on their lipophilicity data. Methods. Chromatographic hydrophobicity data (parameter C0 were obtained on cellulose layers under conditions of normal-phase thin-layer chromatography (NPTLC, using different binary solvent systems. The ACE inhibitors lipophilicity descriptors (logP values were calculated using the software package Virtual Computational Chemistry Laboratory. The ACE inhibitors plasma protein binding data were collected from relevant literature. Results. ACE inhibitors protein binding data varied from negligible (lisinopril to 99% (fosinopril. The calculated lipophilicity descriptors, logPKOWWIN values ranged from -0.94 (lisinopril to 6.61 (fosinopril. Good correlations were established between plasma protein binding values and calculated logPKOWWIN values (R2=0.8026 as well as chromatographic hydrophobicity data, C0 parameters (R2=0.7662. Even though good correlation coefficients (R2 were obtained in both relations, unacceptable probability value with p>0.05 was found in relation between protein binding data and calculated logPKOWWIN values. Subsequently, taking into consideration the request for probability value lower than 0.05, a better relationship was observed between protein binding data and chromatographically obtained hydrophobicity parameters C0 values. Conclusion. Cellulose layers are easily available and cost effective sorbent to assess hydrophobicity. Experimentally obtained data on ACE inhibitors hydrophobicity and plasma protein binding estimation are important parameters in evaluating bioavailability of these drugs. [Projekat Ministarstva

  13. EDITORIAL: Plasma jets and plasma bullets Plasma jets and plasma bullets

    Science.gov (United States)

    Kong, M. G.; Ganguly, B. N.; Hicks, R. F.

    2012-06-01

    Plasma plumes, or plasma jets, belong to a large family of gas discharges whereby the discharge plasma is extended beyond the plasma generation region into the surrounding ambience, either by a field (e.g. electromagnetic, convective gas flow, or shock wave) or a gradient of a directionless physical quantity (e.g. particle density, pressure, or temperature). This physical extension of a plasma plume gives rise to a strong interaction with its surrounding environment, and the interaction alters the properties of both the plasma and the environment, often in a nonlinear and dynamic fashion. The plasma is therefore not confined by defined physical walls, thus extending opportunities for material treatment applications as well as bringing in new challenges in science and technology associated with complex open-boundary problems. Some of the most common examples may be found in dense plasmas with very high dissipation of externally supplied energy (e.g. in electrical, optical or thermal forms) and often in or close to thermal equilibrium. For these dense plasmas, their characteristics are determined predominantly by strong physical forces of different fields, such as electrical, magnetic, thermal, shock wave, and their nonlinear interactions [1]. Common to these dense plasma plumes are significant macroscopic plasma movement and considerable decomposition of solid materials (e.g. vaporization). Their applications are numerous and include detection of elemental traces, synthesis of high-temperature materials and welding, laser--plasma interactions, and relativistic jets in particle accelerators and in space [2]-[4]. Scientific challenges in the understanding of plasma jets are exciting and multidisciplinary, involving interweaving transitions of all four states of matter, and their technological applications are wide-ranging and growing rapidly. Using the Web of Science database, a search for journal papers on non-fusion plasma jets reveals that a long initial phase up

  14. [Acute plasma cell leukemia].

    Science.gov (United States)

    Monsalbe, V; Domíngues, C; Roa, I; Busel, D; González, S

    1989-01-01

    Plasma Cell Leukemia is a very rare form of plasmocytic dyscrasia, whose clinical and pathological characteristics warrant its recognition as a distinct subentity. We report the case of a 60 years old man who presented a rapidly fatal acute plasma cell leukemia, with multiple osteolytic lesions, hipercalcemia, renal and cardiac failure.

  15. Plasma polarization spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Fujimoto, Takashi [Department of Engineering Physics and Mechanics, Graduate School of Engineering, Kyoto University, Kyoto (Japan)

    2000-03-01

    Polarization of radiation emitted from a plasma reflects the anisotropic properties of the plasma, especially the angular anisotropic distribution of electron velocities. Polarization has been observed on impurity ion lines from the WT-3 tokamak and the GAMMA-10 tandem mirror machines. The soft x-ray laser line from the neonlike germanium was also found polarized. (author)

  16. Plasma polarization spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Iwamae, Atsushi; Inoue, Takeru; Tanaka, Akihiro; Kawakami, Kazuki; Fujimoto, Takashi [Kyoto Univ., Dept. of Engineering Physics, Kyoto (Japan)

    2000-03-01

    Polarization of radiation emitted from plasma reflects the anisotropic properties of the plasma, especially the angular isotropic distribution of electron velocities. Polarization has been observed on impurity ion lines from the WT-3 tokamak and GAMMA 10 tandem mirror device. (author)

  17. Atoms in dense plasmas

    Energy Technology Data Exchange (ETDEWEB)

    More, R.M.

    1986-01-01

    Recent experiments with high-power pulsed lasers have strongly encouraged the development of improved theoretical understanding of highly charged ions in a dense plasma environment. This work examines the theory of dense plasmas with emphasis on general rules which govern matter at extreme high temperature and density. 106 refs., 23 figs.

  18. Plasma etching an introduction

    CERN Document Server

    Manos, Dennis M

    1989-01-01

    Plasma etching plays an essential role in microelectronic circuit manufacturing. Suitable for researchers, process engineers, and graduate students, this book introduces the basic physics and chemistry of electrical discharges and relates them to plasma etching mechanisms. Throughout the volume the authors offer practical examples of process chemistry, equipment design, and production methods.

  19. Modelling of Complex Plasmas

    NARCIS (Netherlands)

    Akdim, M.R. (Mohamed Reda)

    2003-01-01

    Nowadays plasmas are used for various applications such as the fabrication of silicon solar cells, integrated circuits, coatings and dental cleaning. In the case of a processing plasma, e.g. for the fabrication of amorphous silicon solar cells, a mixture of silane and hydrogen gas is injected in a r

  20. "Angular" plasma cell cheilitis.

    Science.gov (United States)

    da Cunha Filho, Roberto Rheingantz; Tochetto, Lucas Baldissera; Tochetto, Bruno Baldissera; de Almeida, Hiram Larangeira; Lorencette, Nádia Aparecida; Netto, José Fillus

    2014-03-17

    Plasma cell cheilitis is an extremely rare disease, characterized by erythematous-violaceous, ulcerated and asymptomatic plaques, which evolve slowly. The histological characteristics include dermal infiltrate composed of mature plasmocytes. We report a case of Plasma cell angular cheilitis in a 58-year-old male, localized in the lateral oral commissure.

  1. "Angular" plasma cell cheilitis

    OpenAIRE

    da Cunha Filho, Roberto Rheingantz; Tochetto, Lucas Baldissera; Tochetto, Bruno Baldissera; de Almeida Jr, Hiram Larangeira; Lorencette, Nadia Aparecida; Netto, Jose Fillus

    2014-01-01

    Plasma cell cheilitis is an extremely rare disease, characterized by erythematous-violaceous, ulcerated and asymptomatic plaques, which evolve slowly. The histological characteristics include dermal infiltrate composed of mature plasmocytes. We report a case of Plasma cell angular cheilitis in a 58-year-old male, localized in the lateral oral commissure.

  2. Introduction to Plasma Spectroscopy

    CERN Document Server

    Kunze, H-J

    2009-01-01

    Based on lectures given at the Ruhr-University of Bochum for graduate students and postgraduates starting in plasma physics as well as from low- to high-density hot plasmas, this book introduces basic ideas and fundamental concepts and typical instrumentation from the X-ray to the infrared spectral regions

  3. Innovations in plasma sensors

    Science.gov (United States)

    Zurbuchen, Thomas H.; Gershman, Daniel J.

    2016-04-01

    During the history of space exploration, ever improving instruments have continued to enable new measurements and discoveries. Focusing on plasma sensors, we examine the processes by which such new instrument innovations have occurred over the past decades. Due to risk intolerance prevalent in many NASA space missions, innovations in plasma instrumentation occur primarily when heritage systems fail to meet science requirements, functional requirements as part of its space platform, or design constraints. We will review such innovation triggers in the context of the design literature and with the help of two case studies, the Fast Imaging Plasma Spectrometer on MErcury Surface, Space ENvironment, GEochemistry, and Ranging and the Fast Plasma Investigation on Magnetosphere Multiscale. We will then discuss the anticipated needs for new plasma instrument innovations to enable the science program of the next decade.

  4. Wakes in inhomogeneous plasmas

    CERN Document Server

    Kompaneets, Roman; Nosenko, Vladimir; Morfill, Gregor E

    2014-01-01

    The Debye shielding of a charge immersed in a flowing plasma is an old classic problem in plasma physics. It has been given renewed attention in the last two decades in view of experiments with complex plasmas, where charged dust particles are often levitated in a region with strong ion flow. Efforts to describe the shielding of the dust particles in such conditions have been focused on the homogeneous plasma approximation, which ignores the substantial inhomogeneity of the levitation region. We address the role of the plasma inhomogeneity by rigorously calculating the point charge potential in the collisionless Bohm sheath. We demonstrate that the inhomogeneity can dramatically modify the wake, making it non-oscillatory and weaker.

  5. Basic plasma physics

    CERN Document Server

    Ghosh, Basudev

    2014-01-01

    Basic Plasma Physics is designed to serve as an introductory compact textbook for advanced undergraduate, postgraduate and research students taking plasma physics as one of their subject of study for the first time. It covers the current syllabus of plasma physics offered by the most universities and technical institutions. The book requires no background in plasma physics but only elementary knowledge of basic physics and mathematics. Emphasis has been given on the analytical approach. Topics are developed from first principle so that the students can learn through self-study. One chapter has been devoted to describe some practical aspects of plasma physics. Each chapter contains a good number of solved and unsolved problems and a variety of review questions, mostly taken from recent examination papers. Some classroom experiments described in the book will surely help students as well as instructors.

  6. Microphysics of cosmic plasmas

    CERN Document Server

    Bykov, Andrei; Cargill, Peter; Dendy, Richard; Wit, Thierry; Raymond, John

    2014-01-01

    This title presents a review of the detailed aspects of the physical processes that underlie the observed properties, structures and dynamics of cosmic plasmas. An assessment of the status of understanding of microscale processes in all astrophysical collisionless plasmas is provided. The topics discussed include  turbulence in astrophysical and solar system plasmas as a phenomenological description of their dynamic properties on all scales; observational, theoretical and modelling aspects of collisionless magnetic reconnection; the formation and dynamics of shock waves; and a review and assessment of microprocesses, such as the hierarchy of plasma instabilities, non-local and non-diffusive transport processes and ionisation and radiation processes.  In addition, some of the lessons that have been learned from the extensive existing knowledge of laboratory plasmas as applied to astrophysical problems are also covered.   This volume is aimed at graduate students and researchers active in the areas of cosmi...

  7. SUPERFAST THERMALIZATION OF PLASMA

    Science.gov (United States)

    Chang, C.C.

    1962-06-12

    A method is given for the superfast thermalization of plasma by shock conversion of the kinetic energy stored in rotating plasma rings or plasmoids colliding at near supersonic speeds in a containment field to heat energy in the resultant confined plasma mass. The method includes means for generating rotating plasmoids at the opposite ends of a Pyrotron or Astron containment field. The plasmoids are magnetically accelerated towards each other into the opposite ends of time containment field. During acceleration of the plasmoids toward the center of the containment field, the intensity of the field is sequentially increased to adiabatically compress the plasmoids and increase the plasma energy. The plasmoids hence collide with a violent shock at the eenter of the containment field, causing the substantial kinetic energy stored in the plasmoids to be converted to heat in the resultant plasma mass. (AEC)

  8. Plasma polymerization by Softplasma

    DEFF Research Database (Denmark)

    Jiang, J.; Wu, Zhenning; Benter, Maike

    2008-01-01

    as reactive splvent (as shown in Figure 1). 1] H. Biederman, in Plasma Polymer Films. (ed.) H. Biederman. Imperial College Press, Singapore, 13-24 ~OO~· '. , [2] R. d'Agostino et.a!. in Plasma Depd~itiqn, 'Treatment, and Etching ofPolymers. (ed.) R. d'Agostino, Academic Press, U.S. (1990). [3] F. F. Shi......In the late 19th century, the first depositions - known today as plasma polymers, were reported. In the last century, more and more research has been put into plasma polymers. Many different deposition systems have been developed. [1, 2] Shi F. F. broadly classified them into internal electrode......, external electrode, and electrodeless microwave or high frequency reactors. [3] Softplasma™ is an internal electrode plasma setup powered by low frequenc~ gower supply. It was developed in late 90s for surface treatment of silicone rubber. [ ]- 5] It is a low pressure, low electron density, 3D homogenous...

  9. Gingival plasma cell granuloma

    Directory of Open Access Journals (Sweden)

    Phadnaik Mangesh

    2010-01-01

    Full Text Available Plasma cell granuloma is a rare reactive lesion composed of polyclonal plasma cells. It manifests primarily in the lungs, but may occur in various other anatomic locations like the oral cavity. Intraoral plasma cell granulomas involving the tongue, lip, oral mucosa and gingiva have been reported in the past. This case presents a 54-year-old female with chronic periodontitis and mandibular anterior gingival overgrowth treated by Phase I therapy (scaling and root planing and excisional biopsy. Histological examination revealed inflammatory cell infiltrate containing sheets of plasma cells. Immunohistochemistry for kappa and lambda light chains showed a polyclonal staining pattern confirming a diagnosis of plasma cell granuloma. This case highlights the need to biopsy for unusual lesions to rule out potential neoplasms.

  10. Helical plasma thruster

    Energy Technology Data Exchange (ETDEWEB)

    Beklemishev, A. D., E-mail: bekl@bk.ru [Budker Institute of Nuclear Physics SB RAS, Novosibirsk (Russian Federation)

    2015-10-15

    A new scheme of plasma thruster is proposed. It is based on axial acceleration of rotating magnetized plasmas in magnetic field with helical corrugation. The idea is that the propellant ionization zone can be placed into the local magnetic well, so that initially the ions are trapped. The E × B rotation is provided by an applied radial electric field that makes the setup similar to a magnetron discharge. Then, from the rotating plasma viewpoint, the magnetic wells of the helically corrugated field look like axially moving mirror traps. Specific shaping of the corrugation can allow continuous acceleration of trapped plasma ions along the magnetic field by diamagnetic forces. The accelerated propellant is expelled through the expanding field of magnetic nozzle. By features of the acceleration principle, the helical plasma thruster may operate at high energy densities but requires a rather high axial magnetic field, which places it in the same class as the VASIMR{sup ®} rocket engine.

  11. Diagnostics of Nanodusty Plasma

    Science.gov (United States)

    Greiner, Franko; Groth, Sebastian; Tadsen, Bejamin; Piel, Alexander

    2015-11-01

    The diagnostic of nanodusty plasmas, i.e. plasmas including nano-sized dust particles, is a challenging task. For both, the diagnostic of the nanodusty plasma itself, and the in-situ diagnostic of the nanoparticles, no standard diagnostic exist. Nanodust particle size and density can be estimated using light scattering techniques, namely kinetic Mie ellipsometry and extinction measurements. The charge of the nanoparticles can be estimated from the analysis of dust density waves (DDW). Parameters like the electron density, which give information about the plasma itself, may be deduced from the DDW analysis. We present detailed investigations on nanodust in a reactive Argon-Acetylene plasma created in an rf-driven parallel plate reactor at low pressure using the above mentioned portfolio of diagnostic. Funded by DFG under contract SFB TR-24/A2.

  12. Solar system plasma waves

    Science.gov (United States)

    Gurnett, Donald A.

    1995-01-01

    An overview is given of spacecraft observations of plasma waves in the solar system. In situ measurements of plasma phenomena have now been obtained at all of the planets except Mercury and Pluto, and in the interplanetary medium at heliocentric radial distances ranging from 0.29 to 58 AU. To illustrate the range of phenomena involved, we discuss plasma waves in three regions of physical interest: (1) planetary radiation belts, (2) planetary auroral acceleration regions and (3) the solar wind. In each region we describe examples of plasma waves that are of some importance, either due to the role they play in determining the physical properties of the plasma, or to the unique mechanism involved in their generation.

  13. Plasma and synovial fluid meclofenamic acid concentrations in patients with rheumatoid arthritis of the knee.

    Science.gov (United States)

    Koup, J R; Thomas, D; Tucker, E; Black, A; Ruderman, M; Dixon, J A; Kinkel, A

    1988-01-01

    We have measured plasma and synovial fluid concentrations of meclofenamic acid at 2, 4, 8, and 12 h during steady-state administration (100 mg three times daily for 4-7 days). Paired plasma and synovial samples were obtained pre-treatment and at one of the above times in twelve patients with a diagnosis of rheumatoid arthritis. In addition, the extent of protein binding of meclofenamic acid was assessed in vitro in the pre-treatment plasma and synovial fluid specimens. Peak total concentrations of 1.73 and 0.86 micrograms.ml-1 were observed in plasma (at 2 h) and synovial fluid (at 4 h) respectively. The extent of protein binding was 99.7 and 99.6% (not significantly different) in plasma and synovial fluid respectively. The results of this study are compared to those from similar reported studies of other nonsteroidal anti-inflamatory compounds.

  14. Electronegative Plasma Instabilities in Industrial Pulsed Plasmas

    Science.gov (United States)

    Pribyl, Patrick; Hansen, Anders; Gekelman, Walter

    2016-10-01

    Electronegative gases that are important for industrial etch processes have a series of instabilities that occur at process relevant conditions. These have been studied since the 1990s, but are becoming a much more important today as plasma reactors are being pushed to produce ever finer features, and tight control of the etch process is becoming crucial. The experiments are being done in a plasma etch tool that closely simulates a working industrial device. ICP coils in different configurations are driven by a pulsed RF generators operating at 2-5 MHz. A computer controlled automated probe drive can access a volume above the substrate. The probe can be a Langmuir probe, a ``Bdot'' probe, or an emissive probe the latter used for more accurate determination of plasma potential. A microwave interferometer is available to measure line-averaged electron density. The negative ion instability is triggered depending upon the gas mix (Ar,SF6) , pressure and RF power. The instability can be ``burned through'' by rapidly pulsing the RF power. In this study we present measurements of plasma current and density distribution over the wafer before, after and during the rapid onset of the instability. Work suported by NSF-GOALI Award and done at the BAPSF.

  15. Study of oxygen plasma pre-treatment of polyester fabric for improved polypyrrole adhesion

    Energy Technology Data Exchange (ETDEWEB)

    Mehmood, Tariq [School of Engineering, Deakin University, Geelong, Victoria 3216 (Australia); Kaynak, Akif, E-mail: akaynak@deakin.edu.au [School of Engineering, Deakin University, Geelong, Victoria 3216 (Australia); Dai, Xiujuan J. [Institute for Frontier Materials, Deakin University, Geelong, Victoria 3216 (Australia); Kouzani, Abbas [School of Engineering, Deakin University, Geelong, Victoria 3216 (Australia); Magniez, Kevin; Rubin de Celis, David; Hurren, Christopher J. [Institute for Frontier Materials, Deakin University, Geelong, Victoria 3216 (Australia); Plessis, Johan du [School of Applied Sciences, RMIT University, GPO Box 2476V, Melbourne, Victoria 3001 (Australia)

    2014-01-15

    In this work, we have systematically studied the improvement of binding of polypyrrole with polyethylene terephthalate (PET) thin films and fabrics using low pressure oxygen plasma. A range of plasma treatment times were employed to investigate plasma induced effects on surface roughness, surface chemistry and hydrophilicity. Modifications of PET films were studied with respect to surface morphology by means of atomic force and scanning electron microscopy. Chemical effects of plasma treatment were studied using X-ray photoelectron spectroscopy. Results showed that both the increase in surface functionalisation by carboxylic groups and formation of nano size roughness contributed to improved adhesion and conductivity. - Highlights: • Improved adhesion between polypyrrole and polyester due to oxygen plasma treatment. • Formation of additional C–O and O–C=O groups due to plasma reactions. • Plasma induced increase in surface energy and roughness.

  16. Asymmetric cation-binding catalysis

    DEFF Research Database (Denmark)

    Oliveira, Maria Teresa; Lee, Jiwoong

    2017-01-01

    and KCN, are selectively bound to the catalyst, providing exceptionally high enantioselectivities for kinetic resolutions, elimination reactions (fluoride base), and Strecker synthesis (cyanide nucleophile). Asymmetric cation-binding catalysis was recently expanded to silicon-based reagents, enabling...... solvents, thus increasing their applicability in synthesis. The expansion of this concept to chiral polyethers led to the emergence of asymmetric cation-binding catalysis, where chiral counter anions are generated from metal salts, particularly using BINOL-based polyethers. Alkali metal salts, namely KF...

  17. Effects of protein binding on the biodistribution of PEGylated PLGA nanoparticles post oral administration

    CSIR Research Space (South Africa)

    Semete, B

    2012-03-01

    Full Text Available detected in plasma was higher than that of uncoated PLGA particles, indicating that systemic circulation time can also be increased with oral formulations. The difference in the in vitro protein binding as a result of the different poloxamers used versus...

  18. Sex Hormone Binding Globulin Deficiency Due to a Homozygous Missense Mutation

    NARCIS (Netherlands)

    Vos, M. J.; Mijnhout, G. S.; Rondeel, J. M. M.; Baron, W.; Groeneveld, P. H. P.

    2014-01-01

    Context: SHBG is known as the major sex steroid binding protein in plasma, and it regulates the bioavailability of both T and estradiol levels required for effects on target tissues. We identified a man with an undetectable SHBG concentration in combination with low total T. He presented with a 7-ye

  19. Effect of fibronectin on the binding of antithrombin III to immobilized heparin

    NARCIS (Netherlands)

    Byun, Youngro; Jacobs, Harvey A.; Feijen, Jan; Kim, Sung Wan

    1996-01-01

    An objective of this research is to verify the mechanism of anticoagulant activity of surface-immobilized heparin in the presence of plasma proteins. The competition and binding interaction between immobilized heparin and antithrombin III (ATIII)/thrombin have been described in vitro. However, the s

  20. A comparison of circulating and regional growth hormone-binding protein in cirrhosis

    DEFF Research Database (Denmark)

    Møller, S; Fisker, S; Becker, U;

    2001-01-01

    The growth hormone (GH)/insulin-like growth factor-I (IGF-I) axis is disturbed in cirrhosis, with elevated basal GH and low IGF-I levels relating to liver function and prognosis. In plasma, GH is bound to a high-affinity GH-binding protein (GHBP), which has been found to be slightly reduced...

  1. Metal-Binding Characteristics of the Gamma-Glutamyl Capsular Polymer of Bacillus licheniformis ATCC 9945.

    Science.gov (United States)

    McLean, R J; Beauchemin, D; Clapham, L; Beveridge, T J

    1990-12-01

    The metal-binding affinity of the anionic poly-gamma-d-glutamyl capsule of Bacillus licheniformis was investigated by using Na, Mg, Al, Ca, Cr, Mn, Fe, Ni, and Cu. Purified capsule was suspended in various concentrations of the chloride salts of the various metals, and after dialysis the bound metals were analyzed either by graphite furnace atomic absorption spectroscopy or by inductively coupled plasma-mass spectrometry. Exposure of purified capsule to excess concentrations of Na revealed it to contain 8.2 mumol of anionic sites per mg on the basis of Na binding. This was confirmed by titration of the capsule with HCl and NaOH. Other metal ions were then added in ionic concentrations equivalent to 25, 50, 75, 100, 200, and 400% of the available anionic sites. The binding characteristics varied with the metal being investigated. Addition of Cu, Al, Cr, or Fe induced flocculation. These metal ions showed the greatest affinity for B. licheniformis capsule in competitive-binding experiments. Flocculation was not seen with the addition of other metal ions. With the exception of Ni and Fe all capsule-metal-binding sites readily saturated. Ni had low affinity for the polymer, and its binding was increased at high metal concentrations. Fe binding resulted in the development of rust-colored ferrihydrite which itself could bind additional metal. Metal-binding characteristics of B. licheniformis capsule appear to be influenced by the chemical and physical properties of both the capsule and the metal ions.

  2. Nonthermal plasma chemistry and physics

    CERN Document Server

    Meichsner, Jurgen; Schneider, Ralf; Wagner, Hans-Erich

    2013-01-01

    In addition to introducing the basics of plasma physics, Nonthermal Plasma Chemistry and Physics is a comprehensive presentation of recent developments in the rapidly growing field of nonthermal plasma chemistry. The book offers a detailed discussion of the fundamentals of plasma chemical reactions and modeling, nonthermal plasma sources, relevant diagnostic techniques, and selected applications.Elucidating interconnections and trends, the book focuses on basic principles and illustrations across a broad field of applications. Expert contributors address environmental aspects of plasma chemist

  3. The large intracellular loop of hZIP4 is an intrinsically disordered zinc binding domain†

    Science.gov (United States)

    Bafaro, Elizabeth M.; Antala, Sagar; Nguyen, Tuong-Vi; Dzul, Stephen P.; Doyon, Brian; Stemmler, Timothy L.; Dempski, Robert E.

    2015-01-01

    The human (h) ZIP4 transporter is a plasma membrane protein which functions to increase the cytosolic concentration of zinc. hZIP4 transports zinc into intestinal cells and therefore has a central role in the absorption of dietary zinc. hZIP4 has eight transmembrane domains and encodes a large intracellular loop between transmembrane domains III and IV, M3M4. Previously, it has been postulated that this domain regulates hZIP4 levels in the plasma membrane in a zinc-dependent manner. The objective of this research was to examine the zinc binding properties of the large intracellular loop of hZIP4. Therefore, we have recombineantly expressed and purified M3M4 and showed that this domain binds two zinc ions. Using a combination of site-directed mutagenesis, metal binding affinity assays, and X-ray absorption spectroscopy, we demonstrated that the two Zn2+ ions bind sequentially, with the first Zn2+ binding to a CysHis3 site with a nanomolar binding affinity, and the second Zn2+ binding to a His4 site with a weaker affinity. Circular dichroism spectroscopy revealed that the M3M4 domain is intrinsically disordered, with only a small structural change induced upon Zn2+ coordination. Our data supports a model in which the intracellular M3M4 domain senses high cytosolic Zn2+ concentrations and regulates the plasma membrane levels of the hZIP4 transporter in response to Zn2+ binding. PMID:25882556

  4. Plasma physics an introduction to laboratory, space, and fusion plasmas

    CERN Document Server

    Piel, Alexander

    2017-01-01

    The enlarged new edition of this textbook provides a comprehensive introduction to the basic processes in plasmas and demonstrates that the same fundamental concepts describe cold gas-discharge plasmas, space plasmas, and hot fusion plasmas. Starting from particle drifts in magnetic fields, the principles of magnetic confinement fusion are explained and compared with laser fusion. Collective processes are discussed in terms of plasma waves and instabilities. The concepts of plasma description by magnetohydrodynamics, kinetic theory, and particle simulation are stepwise introduced. Space charge effects in sheath regions, double layers and plasma diodes are given the necessary attention. The novel fundamental mechanisms of dusty plasmas are explored and integrated into the framework of conventional plasmas. The book concludes with a concise description of modern plasma discharges. Written by an internationally renowned researcher in experimental plasma physics, the text keeps the mathematical apparatus simple a...

  5. The heparin-binding site in tetranectin is located in the N-terminal region and binding does not involve the carbohydrate recognition domain

    DEFF Research Database (Denmark)

    Lorentsen, R H; Graversen, Jonas Heilskov; Caterer, N R

    2000-01-01

    in three exons. Exon 3 encodes the carbohydrate recognition domain, which binds to kringle 4 in plasminogen at low levels of Ca(2+). Exon 2 encodes an alpha-helix, which is necessary and sufficient to govern the trimerization of tetranectin by assembling into a triple-helical coiled-coil structural element......Tetranectin is a homotrimeric plasma and extracellular-matrix protein that binds plasminogen and complex sulphated polysaccharides including heparin. In terms of primary and tertiary structure, tetranectin is related to the collectin family of Ca(2+)-binding C-type lectins. Tetranectin is encoded....... Here we show that the heparin-binding site in tetranectin resides not in the carbohydrate recognition domain but within the N-terminal region, comprising the 16 amino acid residues encoded by exon 1. In particular, the lysine residues in the decapeptide segment KPKKIVNAKK (tetranectin residues 6...

  6. Evidence for the presence of lipid-free monomolecular apolipoprotein A-1 in plasma[S

    Science.gov (United States)

    Miyazaki, Osamu; Ogihara, Jun; Fukamachi, Isamu; Kasumi, Takafumi

    2014-01-01

    The first step in reverse cholesterol transport is a process by which lipid-free or lipid-poor apoA-1 removes cholesterol from cells through the action of ATP binding cassette transporter A1 at the plasma membrane. However the structure and composition of lipid-free or -poor apoA-1 in plasma remains obscure. We previously obtained a monoclonal antibody (MAb) that specifically recognizes apoA-1 in preβ1-HDL, the smallest apoA-1-containing particle in plasma, which we used to establish a preβ1-HDL ELISA. Here, we purified preβ1-HDL from fresh normal plasma using said antibody, and analyzed the composition and structure. ApoA-1 was detected, but neither phospholipid nor cholesterol were detected in the purified preβ1-HDL. Only globular, not discoidal, particles were observed by electron microscopy. In nondenaturing PAGE, no difference in the mobility was observed between the purified preβ1-HDL and original plasma preβ1-HDL, or between the preβ1-HDL and lipid-free apoA-1 prepared by delipidating HDL. In sandwich ELISA using two anti-preβ1-HDL MAbs, reactivity with intact plasma preβ1-HDL was observed in ELISA using two MAbs with distinct epitopes but no reactivity was observed in ELISA using a single MAb, and the same phenomenon was observed with monomolecular lipid-free apoA-1. These results suggest that plasma preβ1-HDL is lipid-free monomolecular apoA-1. PMID:24304668

  7. Calcium-binding proteins from human platelets

    Energy Technology Data Exchange (ETDEWEB)

    Gogstad, G.O.; Krutnes, M.B.; Solum, N.O.

    1983-06-01

    Calcium-binding platelet proteins were examined by crossed immunoelectrophoresis of solubilized platelets against antibodies to whole platelets followed by incubation of the immunoplates with /sup 45/Ca/sup 2 +/ and autoradiography. When the immunoplates had been pretreated with EDTA at pH 9.0 in order to remove divalent cations, three immunoprecipitates were markedly labelled with /sup 45/Ca/sup 2 +/. These corresponded to the glycoprotein IIb-IIIa complex, glycoprotein Ia and a presently unidentified antigen termed G18. These antigens were membrane-bound and surface-oriented. When an excess of EDTA was introduced in the incubation media the results revealed that the glycoprotein IIb-IIIa complex and antigen G18, but not glycoprotein Ia, contained sites with a stronger affinity for calcium than has EDTA at pH 7.4. Immunoprecipitates of the separate glycoproteins IIb and IIIa both bound calcium in the same manner as the glycoprotein IIb-IIIa complex. As another approach, platelet-rich plasma was incubated with /sup 45/Ca/sup 2 +/ prior to crossed immunoelectrophoresis of the solubilized platelets. A single immunoprecipitate was weakly labelled. This did not correspond to any of the immunoprecipitates which were visible after staining with Coomassie blue. The labelling of this antigen was markedly increased when the platelet-rich plasma had been preincubated with EDTA and in this case a weak labelling of the glycoprotein IIB-IIIa precipitate also became apparent. No increased incorporation of calcium occured in any of these immunoprecipitates when the platelets were aggregated with ADP in the presence of /sup 45/Ca/sup 2 +/.

  8. Mapping cocaine binding sites in human and baboon brain in vivo.

    Science.gov (United States)

    Fowler, J S; Volkow, N D; Wolf, A P; Dewey, S L; Schlyer, D J; Macgregor, R R; Hitzemann, R; Logan, J; Bendriem, B; Gatley, S J

    1989-01-01

    The first direct measurements of cocaine binding in the brain of normal human volunteers and baboons have been made by using positron emission tomography (PET) and tracer doses of [N-11C-methyl]-(-)-cocaine ([11C]cocaine). Cocaine's binding and release from brain are rapid with the highest regional uptake of carbon-11 occurring in the corpus striatum at 4-10 minutes after intravenous injection of labeled cocaine. This was followed by a clearance to half the peak value at about 25 minutes with the overall time course paralleling the previously documented time course of the euphoria experienced after intravenous cocaine administration. Blockade of the dopamine reuptake sites with nomifensine reduced the striatal but not the cerebellar uptake of [11C]cocaine in baboons indicating that cocaine binding is associated with the dopamine reuptake site in the corpus striatum. A comparison of labeled metabolites of cocaine in human and baboon plasma showed that while cocaine is rapidly metabolized in both species, the profile of labeled metabolites is different, with baboon plasma containing significant amounts of labeled carbon dioxide, and human plasma containing no significant labeled carbon dioxide. These studies demonstrate the feasibility of using [11C]cocaine and PET to map binding sites for cocaine in human brain, to monitor its kinetics, and to characterize its binding mechanism by using appropriate pharmacological challenges.

  9. Early results of microwave transmission experiments through an overly dense rectangular plasma sheet with microparticle injection

    Science.gov (United States)

    Gillman, Eric D.; Amatucci, W. E.

    2014-06-01

    These experiments utilize a linear hollow cathode to create a dense, rectangular plasma sheet to simulate the plasma layer surrounding vehicles traveling at hypersonic velocities within the Earth's atmosphere. Injection of fine dielectric microparticles significantly reduces the electron density and therefore lowers the electron plasma frequency by binding a significant portion of the bulk free electrons to the relatively massive microparticles. Measurements show that microwave transmission through this previously overly dense, impenetrable plasma layer increases with the injection of alumina microparticles approximately 60 μm in diameter. This method of electron depletion is a potential means of mitigating the radio communications blackout experienced by hypersonic vehicles.

  10. Urine and plasma propranolol.

    Science.gov (United States)

    Andreasen, F; Jakobsen, P; Kornerup, H J; Pedersen, E B; Pedersen, O L

    1983-01-01

    Eight hypertensive patients who had been followed in an outpatient clinic during long-term therapy with propranolol (40 to 160 mg twice daily) were studied during a 24-hr stay in the ward. The usual oral dose was given and the total and free plasma concentrations were determined during the 24 hr and the urinary excretion of unchanged drug was measured. Average free plasma concentration of propranolol (y free) was calculated from: y free = Excreted propranolol (ng/24 hr)/Creatinine clearance (ml/24 hr). There was a significant relationship between log y free and average free plasma concentration (means free) determined from the directly measured plasma concentration curve: log y free = 0.0743 means free - 0.0466 (r = 0.98, P less than 0.001). In another group of propranolol-treated hypertensive patients there was a significant positive relationship between orosomucoid concentration and reciprocal of the free propranolol fraction in plasma. From this relationship the average total drug concentration (y total) was calculated from y free; there was a significant correlation with directly measured total plasma level: log y total = 0.0038 . means total + 1.0895 (r = 0.91, P less than 0.001). It is suggested that individually determined values of y free below 30 ng/ml and y total below 400 ng/ml (the concentration range studied) can be used to calculate the average mean 24-hr free and total plasma concentrations.

  11. Dense Hypervelocity Plasma Jets

    Science.gov (United States)

    Case, Andrew; Witherspoon, F. Douglas; Messer, Sarah; Bomgardner, Richard; Phillips, Michael; van Doren, David; Elton, Raymond; Uzun-Kaymak, Ilker

    2007-11-01

    We are developing high velocity dense plasma jets for fusion and HEDP applications. Traditional coaxial plasma accelerators suffer from the blow-by instability which limits the mass accelerated to high velocity. In the current design blow-by is delayed by a combination of electrode shaping and use of a tailored plasma armature created by injection of a high density plasma at a few eV generated by arrays of capillary discharges or sparkgaps. Experimental data will be presented for a complete 32 injector gun system built for driving rotation in the Maryland MCX experiment, including data on penetration of the plasma jet through a magnetic field. We present spectroscopic measurements of plasma velocity, temperature, and density, as well as total momentum measured using a ballistic pendulum. Measurements are in agreement with each other and with time of flight data from photodiodes and a multichannel PMT. Plasma density is above 10^15 cm-3, velocities range up to about 100 km/s. Preliminary results from a quadrature heterodyne HeNe interferometer are consistent with these results.

  12. Plasma Science Committee (PLSC)

    Science.gov (United States)

    1990-12-01

    The Plasma Science Committee (PLSC) is a standing committee under the auspices of the Board on Physics and Astronomy, Commission on Physical Sciences, Mathematics, and Applications of the National Academy of Sciences - National Research Council. Plasma sciences represent a broad and diverse field. The PLSC has accepted the responsibility of monitoring the continuing development and assessing the general health of the field as whole. Although select advisory bodies have been created to address specific issues that affect plasma science, such as the Fusion Policy Advisory Committee (FPAC), the PLSC provides a focus for the plasma science community that is unique and essential. The membership of the PLSC is drawn from research laboratories in universities, industry, and government. Areas of expertise on the committee include accelerators and beams, space physics, astrophysics, computational physics and applied mathematics, fusion plasmas, fundamental experiments and theory, radiation sources, low temperature plasmas, and plasma-surface interactions. The PLSC is well prepared to respond to requests for studies on specific issues.

  13. Influence of plasma density and plasma sheath dynamics on the ion implantation by plasma immersion technique

    OpenAIRE

    Ensinger, Wolfgang

    1996-01-01

    Influence of plasma density and plasma sheath dynamics on the ion implantation by plasma immersion technique / B. Rauschenbach ... - In: Nuclear instruments and methods in physics research. B. 113. 1996. S. 266-269

  14. Tafamidis, a potent and selective transthyretin kinetic stabilizer that inhibits the amyloid cascade

    OpenAIRE

    Bulawa, Christine E.; Connelly, Stephen; DeVit, Michael; Wang, Lan; Weigel, Charlotte; Fleming, James A.; Packman, Jeff; Powers, Evan T.; Wiseman, R Luke; Foss, Theodore R.; Wilson, Ian A.; Kelly, Jeffery W.; Labaudinière, Richard

    2012-01-01

    The transthyretin amyloidoses (ATTR) are invariably fatal diseases characterized by progressive neuropathy and/or cardiomyopathy. ATTR are caused by aggregation of transthyretin (TTR), a natively tetrameric protein involved in the transport of thyroxine and the vitamin A–retinol-binding protein complex. Mutations within TTR that cause autosomal dominant forms of disease facilitate tetramer dissociation, monomer misfolding, and aggregation, although wild-type TTR can also form amyloid fibrils ...

  15. Receptor-binding radiotracers: a class of potential radiopharmaceuticals

    Energy Technology Data Exchange (ETDEWEB)

    Eckelman, W.C.; Reba, R.C.; Gibson, R.E.; Rzeszotarski, W.J.; Vieras, F.; Mazaitis, J.K.; Francis, B.

    1979-04-01

    To date no radiopharmaceutical is routinely used to study changes in receptor concentration. Frequently changes in receptor concentration, or the appearance of receptors in tumors, indicates a specific pathologic state. With a receptor-binding radiotracer, in vivo studies of these changes will be possible. A reversible bimolecular model and in vitro tests were used to determine equilibrium constants and maximal target-to-blood ratios for new derivatives. Theoretical calculations showed that derivatives binding to the estrogen receptor, the beta adrenoceptor, or the cholinergic receptor are capable of achieving satisfactory target-to-blood ratios. Using in vitro tests, the apparent affinity constant was determined for five iodinated estrogen derivatives and five derivatives of beta blockers. Results of the in vitro study with derivatives of beta blockers. Results of the in vitro study with derivatives of beta blockers, and in vivo displacement studies using propranolol, indicated that the high heart-to-blood ratios (5 to 20) obtained with the new derivatives were not the result of a specific interaction with the receptor. In this instance factors other than receptor binding control the in vivo distribution. The in vitro assay using estrogen receptors showed that of the five derivatives, iodohexestrol and 17-alpha-iodoethynylestradiol bind to the receptor with the highest affinity. In vivo studies confirmed these results; iodohexestrol gave a uterus-to-blood ratio of 10 in immature rats when plasma-protein binding was blocked. With a tritiated muscarinic cholinergic blocking agent, heart-to-blood ratios near the theoretical maximum were obtained. This compound most closely follows the mechanism described by the model. Use of the theoretical model in conjunction with in vitro assays can greatly aid in the design of this new class of receptor-binding radiopharmaceuticals.

  16. Stirring Unmagnetized Plasma

    CERN Document Server

    Collins, C; Wallace, J; Jara-Almonte, J; Reese, I; Zweibel, E; Forest, C B; 10.1103/PhysRevLett.108.115001

    2012-01-01

    A new concept for spinning unmagnetized plasma is demonstrated experimentally. Plasma is confined by an axisymmetric multi-cusp magnetic field and biased cathodes are used to drive currents and impart a torque in the magnetized edge. Measurements show that flow viscously couples momentum from the magnetized edge (where the plasma viscosity is small) into the unmagnetized core (where the viscosity is large) and that the core rotates as a solid body. To be effective, collisional viscosity must overcome the ion-neutral drag due to charge exchange collisions.

  17. Optical plasma microelectronic devices