WorldWideScience

Sample records for stable protein concentrates

  1. Concentration of stable elements in food products

    International Nuclear Information System (INIS)

    Montford, M.A.; Shank, K.E.; Hendricks, C.; Oakes, T.W.

    1980-01-01

    Food samples were taken from commercial markets and analyzed for stable element content. The concentrations of most stable elements (Ag, Al, As, Au, Ba, Br, Ca, Ce, Cl, Co, Cr, Cs, Cu, Fe, Hf, I, K, La, Mg, Mn, Mo, Na, Rb, Sb, Sc, Se, Sr, Ta, Th, Ti, V, Zn, Zr) were determined using multiple-element neutron activation analysis, while the concentrations of other elements (Cd, Hg, Ni, Pb) were determined using atomic absorption. The relevance of the concentrations found are noted in relation to other literature values. An earlier study was extended to include the determination of the concentration of stable elements in home-grown products in the vicinity of the Oak Ridge National Laboratory. Comparisons between the commercial and local food-stuff values are discussed

  2. Protein-based stable isotope probing.

    Science.gov (United States)

    Jehmlich, Nico; Schmidt, Frank; Taubert, Martin; Seifert, Jana; Bastida, Felipe; von Bergen, Martin; Richnow, Hans-Hermann; Vogt, Carsten

    2010-12-01

    We describe a stable isotope probing (SIP) technique that was developed to link microbe-specific metabolic function to phylogenetic information. Carbon ((13)C)- or nitrogen ((15)N)-labeled substrates (typically with >98% heavy label) were used in cultivation experiments and the heavy isotope incorporation into proteins (protein-SIP) on growth was determined. The amount of incorporation provides a measure for assimilation of a substrate, and the sequence information from peptide analysis obtained by mass spectrometry delivers phylogenetic information about the microorganisms responsible for the metabolism of the particular substrate. In this article, we provide guidelines for incubating microbial cultures with labeled substrates and a protocol for protein-SIP. The protocol guides readers through the proteomics pipeline, including protein extraction, gel-free and gel-based protein separation, the subsequent mass spectrometric analysis of peptides and the calculation of the incorporation of stable isotopes into peptides. Extraction of proteins and the mass fingerprint measurements of unlabeled and labeled fractions can be performed in 2-3 d.

  3. Protein labelling with stable isotopes: strategies

    International Nuclear Information System (INIS)

    Lirsac, P.N.; Gilles, N.; Jamin, N.; Toma, F.; Gabrielsen, O.; Boulain, J.C.; Menez, A.

    1994-01-01

    A protein labelling technique with stable isotopes has been developed at the CEA: a labelled complete medium has been developed, performing as well as the Luria medium, but differing from it because it contains not only free aminated acids and peptides, but also sugars (96% of D-glucopyrannose) and labelled nucleosides. These precursors are produced from a labelled photosynthetic micro-organisms biomass, obtained with micro-algae having incorporated carbon 13, nitrogen 15 and deuterium during their culture. Labelling costs are reduced. 1 fig., 1 tab., 3 refs

  4. Formulation of stable protein powders by supercritical fluid drying

    NARCIS (Netherlands)

    Jovanović, N.

    2007-01-01

    Protein pharmaceuticals are potent drugs for the treatment of several chronic and life-threatening diseases. However, the complex and sensitive nature of protein molecules requires special attention in the development of stable dosage forms. Developing stable aqueous protein formulations is often a

  5. Radionuclides and selected trace elements in marine protein concentrates

    Energy Technology Data Exchange (ETDEWEB)

    Beasley, T M; Jokela, T A; Eagle, R J

    1971-12-01

    The concentrations of various trace elements and radionuclides have been measured in marine protein concentrates prepared from surface feeding fishes. As with concentrates prepared from benthic fishes, the /sup 210/Pb-/sup 210/Po pair are the most significant radionuclides present. Concentrations of stable Pb, Co and Ag in certain concentrates are sufficiently high to contribute substantially to estimated current intakes of these elements.

  6. A model to secure a stable iodine concentration in milk

    Directory of Open Access Journals (Sweden)

    Gisken Trøan

    2015-12-01

    Full Text Available Background: Dairy products account for approximately 60% of the iodine intake in the Norwegian population. The iodine concentration in cow's milk varies considerably, depending on feeding practices, season, and amount of iodine and rapeseed products in cow fodder. The variation in iodine in milk affects the risk of iodine deficiency or excess in the population. Objective: The first goal of this study was to develop a model to predict the iodine concentration in milk based on the concentration of iodine and rapeseed or glucosinolate in feed, as a tool to securing stable iodine concentration in milk. A second aim was to estimate the impact of different iodine levels in milk on iodine nutrition in the Norwegian population. Design: Two models were developed on the basis of results from eight published and two unpublished studies from the past 20 years. The models were based on different iodine concentrations in the fodder combined with either glucosinolate (Model 1 or rapeseed cake/meal (Model 2. To illustrate the impact of different iodine concentrations in milk on iodine intake, we simulated the iodine contribution from dairy products in different population groups based on food intake data in the most recent dietary surveys in Norway. Results: The models developed could predict iodine concentration in milk. Cross-validation showed good fit and confirmed the explanatory power of the models. Our calculations showed that dairy products with current iodine level in milk (200 µg/kg cover 68, 49, 108 and 56% of the daily iodine requirements for men, women, 2-year-old children, and pregnant women, respectively. Conclusions: Securing a stable level of iodine in milk by adjusting iodine concentration in different cow feeds is thus important for preventing excess intake in small children and iodine deficiency in pregnant and non-pregnant women.

  7. Perioperative plasma concentrations of stable nitric oxide products are predictive of cognitive dysfunction after laparoscopic cholecystectomy.

    LENUS (Irish Health Repository)

    Iohom, G

    2012-02-03

    In this study our objectives were to determine the incidence of postoperative cognitive dysfunction (POCD) after laparoscopic cholecystectomy under sevoflurane anesthesia in patients aged >40 and <85 yr and to examine the associations between plasma concentrations of i) S-100beta protein and ii) stable nitric oxide (NO) products and POCD in this clinical setting. Neuropsychological tests were performed on 42 ASA physical status I-II patients the day before, and 4 days and 6 wk after surgery. Patient spouses (n = 13) were studied as controls. Cognitive dysfunction was defined as deficit in one or more cognitive domain(s). Serial measurements of serum concentrations of S-100beta protein and plasma concentrations of stable NO products (nitrate\\/nitrite, NOx) were performed perioperatively. Four days after surgery, new cognitive deficit was present in 16 (40%) patients and in 1 (7%) control subject (P = 0.01). Six weeks postoperatively, new cognitive deficit was present in 21 (53%) patients and 3 (23%) control subjects (P = 0.03). Compared with the "no deficit" group, patients who demonstrated a new cognitive deficit 4 days postoperatively had larger plasma NOx at each perioperative time point (P < 0.05 for each time point). Serum S-100beta protein concentrations were similar in the 2 groups. In conclusion, preoperative (and postoperative) plasma concentrations of stable NO products (but not S-100beta) are associated with early POCD. The former represents a potential biochemical predictor of POCD.

  8. Cytocompatible and water stable ultrafine protein fibers for tissue engineering

    Science.gov (United States)

    Jiang, Qiuran

    This dissertation proposal focuses on the development of cytocompatible and water stable protein ultrafine fibers for tissue engineering. The protein-based ultrafine fibers have the potential to be used for biomedicine, due to their biocompatibility, biodegradability, similarity to natural extracellular matrix (ECM) in physical structure and chemical composition, and superior adsorption properties due to their high surface to volume ratio. However, the current technologies to produce the protein-based ultrafine fibers for biomedical applications still have several problems. For instance, the current electrospinning and phase separation technologies generate scaffolds composed of densely compacted ultrafine fibers, and cells can spread just on the surface of the fiber bulk, and hardly penetrate into the inner sections of scaffolds. Thus, these scaffolds can merely emulate the ECM as a two dimensional basement membrane, but are difficult to mimic the three dimensional ECM stroma. Moreover, the protein-based ultrafine fibers do not possess sufficient water stability and strength for biomedical applications, and need modifications such as crosslinking. However, current crosslinking methods are either high in toxicity or low in crosslinking efficiency. To solve the problems mentioned above, zein, collagen, and gelatin were selected as the raw materials to represent plant proteins, animal proteins, and denatured proteins in this dissertation. A benign solvent system was developed specifically for the fabrication of collagen ultrafine fibers. In addition, the gelatin scaffolds with a loose fibrous structure, high cell-accessibility and cell viability were produced by a novel ultralow concentration phase separation method aiming to simulate the structure of three dimensional (3D) ECM stroma. Non-toxic crosslinking methods using citric acid as the crosslinker were also developed for electrospun or phase separated scaffolds from these three proteins, and proved to be

  9. Recombinant protein production from stable mammalian cell lines and pools.

    Science.gov (United States)

    Hacker, David L; Balasubramanian, Sowmya

    2016-06-01

    We highlight recent developments for the production of recombinant proteins from suspension-adapted mammalian cell lines. We discuss the generation of stable cell lines using transposons and lentivirus vectors (non-targeted transgene integration) and site-specific recombinases (targeted transgene integration). Each of these methods results in the generation of cell lines with protein yields that are generally superior to those achievable through classical plasmid transfection that depends on the integration of the transfected DNA by non-homologous DNA end-joining. This is the main reason why these techniques can also be used for the generation of stable cell pools, heterogenous populations of recombinant cells generated by gene delivery and genetic selection without resorting to single cell cloning. This allows the time line from gene transfer to protein production to be reduced. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Spontaneous variability of pre-dialysis concentrations of uremic toxins over time in stable hemodialysis patients.

    Directory of Open Access Journals (Sweden)

    Sunny Eloot

    Full Text Available Numerous outcome studies and interventional trials in hemodialysis (HD patients are based on uremic toxin concentrations determined at one single or a limited number of time points. The reliability of these studies however entirely depends on how representative these cross-sectional concentrations are. We therefore investigated the variability of predialysis concentrations of uremic toxins over time.Prospectively collected predialysis serum samples of the midweek session of week 0, 1, 2, 3, 4, 8, 12, and 16 were analyzed for a panel of uremic toxins in stable chronic HD patients (N = 18 while maintaining dialyzer type and dialysis mode during the study period.Concentrations of the analyzed uremic toxins varied substantially between individuals, but also within stable HD patients (intra-patient variability. For urea, creatinine, beta-2-microglobulin, and some protein-bound uremic toxins, Intra-class Correlation Coefficient (ICC was higher than 0.7. However, for phosphorus, uric acid, symmetric and asymmetric dimethylarginine, and the protein-bound toxins hippuric acid and indoxyl sulfate, ICC values were below 0.7, implying a concentration variability within the individual patient even exceeding 65% of the observed inter-patient variability.Intra-patient variability may affect the interpretation of the association between a single concentration of certain uremic toxins and outcomes. When performing future outcome and interventional studies with uremic toxins other than described here, one should quantify their intra-patient variability and take into account that for solutes with a large intra-patient variability associations could be missed.

  11. Directed evolution of an extremely stable fluorescent protein.

    Science.gov (United States)

    Kiss, Csaba; Temirov, Jamshid; Chasteen, Leslie; Waldo, Geoffrey S; Bradbury, Andrew R M

    2009-05-01

    In this paper we describe the evolution of eCGP123, an extremely stable green fluorescent protein based on a previously described fluorescent protein created by consensus engineering (CGP: consensus green protein). eCGP123 could not be denatured by a standard thermal melt, preserved almost full fluorescence after overnight incubation at 80 degrees C and possessed a free energy of denaturation of 12.4 kcal/mol. It was created from CGP by a recursive process involving the sequential introduction of three destabilizing heterologous inserts, evolution to overcome the destabilization and finally 'removal' of the destabilizing insert by gene synthesis. We believe that this approach may be generally applicable to the stabilization of other proteins.

  12. Protein Stable Isotope Fingerprinting (P-SIF): Multidimensional Protein Chromatography Coupled to Stable Isotope-Ratio Mass Spectrometry

    Science.gov (United States)

    Pearson, A.; Bovee, R. J.; Mohr, W.; Tang, T.

    2012-12-01

    As metagenomics increases our insight into microbial community diversity and metabolic potential, new approaches are required to determine the biogeochemical expression of this potential within ecosystems. Because stable isotopic analysis of the major bioactive elements (C, N) has been used historically to map flows of substrates and energy among macroscopic food webs, similar principles may apply to microbes. To address this challenge, we have developed a new analytical approach called Protein Stable Isotope Fingerprinting (P-SIF). P-SIF generates natural stable isotopic fingerprints of microbial individual or community proteomes. The main advantage of P-SIF is the potential to bridge the gap between diversity and function, thereby providing a window into the "black box" of environmental microbiology and helping to decipher the roles of uncultivated species. Our method implements a three-way, orthogonal scheme to separate mixtures of whole proteins into subfractions dominated by single or closely-related proteins. Protein extracts first are isoelectrically focused in a gel-free technique that yields 12 fractions separated over a gradient of pH 3-10. Each fraction then is separated by size-exclusion chromatography into 20 pools, ranging from >100kD to ~10kD. Finally, each of these pools is subjected to HPLC and collected in 40 time-slices based on protein hydrophobicity. Theoretical calculation reveals that the true chromatographic resolution of the total scheme is 5000, somewhat less than the 9600 resulting fractions. High-yielding fractions are subjected to δ13C analysis by spooling-wire microcombustion irMS (SWiM-irMS) optimized for samples containing 1-5 nmol carbon. Here we will present the method, results for a variety of pure cultures, and preliminary data for a sample of mixed environmental proteins. The data show the promise of this method for unraveling the metabolic complexity hidden within microbial communities.

  13. RHEOLOGY OF CHICKPEA PROTEIN CONCENTRATE DISPERSIONS

    Directory of Open Access Journals (Sweden)

    Aurelia Ionescu

    2011-12-01

    Full Text Available Chickpea proteins are used as ingredients in comminuted sausage products and many oriental textured foods. Rheological behaviour of chickpea protein concentrate was studied using a controlled stress rheometer. The protein dispersion prepared with phosphate buffer at pH 7.0 presented non-Newtonian shear thinning behaviour and rheological data well fitted to the Sisko, Carreau and Cross models. The viscoelastic properties of the chickpea protein suspensions were estimated by measuring the storage and loss moduli in oscillatory frequency conditions (0.1-10 Hz at 20°C. Moreover, thermally induced gelation of the chickpea proteins (16, 24 and 36% was studied at pH 7.0 and 4.5 in the temperature range 50 to 100oC and salt concentration ranging from 0 to 1 M. Gelling behaviour was quantified by means of dynamic rheological measurements. Gels formation was preceded by the decrease of storage modulus and loss moduli, coupled with the increase of the phase angle (delta. The beginning of thermal gelation was influenced by protein concentration, pH and salt level. In all studied cases, storage modulus increased rapidly in the temperature range 70-90°C. All rheological parameters measured at 90°C were significantly higher at pH 4.5 compared to pH 7.0.

  14. Protein Concentrate Production from Thin Stillage.

    Science.gov (United States)

    Ratanapariyanuch, Kornsulee; Shim, Youn Young; Emami, Shahram; Reaney, Martin J T

    2016-12-21

    Two-stage fermentation (TSF) of saccharified wheat with a consortium of endemic lactobacilli produced CO 2 and induced colloid separation of fermented solution to produce a protein concentrate (PC). Protein-rich slurry (50%, db) was obtained by decanting solution or skimming floating material during or after TSF. Washing and drying processes were explored to improve protein content, extend storage life of slurry, and yield converted stillage for compound recovery. Centrifuging and washing slurry afforded a PC and clarified solution. PC protein content increased to 60% (w/w, db). The PC was dried in a spray dryer or drum dryer or tray dryer. Dried PC water activity ranged 0.23-0.30. The dried PC lysine content was low, but lysine availability (95%) was excellent. Liquid from TSF and washing was readily microfiltered. Mass recovery of protein, glycerol, 1,3-propanediol, lactic acid, acetic acid, and glycerylphosphorylcholine from combined TSF, washing, and filtration were 66, 76, 72, 77, 74, and 84%, respectively.

  15. Induction of stable protein-deoxyribonucleic acid adducts in Chinese hamster cell chromatin by ultraviolet light

    International Nuclear Information System (INIS)

    Strniste, G.F.; Rall, S.C.

    1976-01-01

    Ultraviolet (uv)-light-mediated formation of protein-DNA adducts in Chinese hamster cell chromatin was investigated in an attempt to compare chromatin alterations induced in vitro with those observed in vivo. Three independent methods of analysis indicated stable protein-DNA associations: a membrane filter assay which retained DNA on the filter in the presence of high salt-detergent; a Sepharose 4B column assay in which protein eluted coincident with DNA; and a CsCl density gradient equilibrium assay which showed both protein and DNA banding at densities other than their respective native densities. Treatment of the irradiated chromatin with DNase provided further evidence that protein--DNA and not protein-protein adducts were being observed in the column assay. There is a fluence-dependent response of protein-DNA adduct formation when the chromatin is irradiated at low ionic strength and is linear for protein over the range studied. When the chromatin is exposed to differing conditions of pH, ionic strength, or divalent metal ion concentration, the quantity of adduct formed upon uv irradiation varies. Susceptibility to adduct formation can be partially explained in terms of the condensation state of the chromatin and other factors such as rearrangement, denaturation, and dissociation of the chromatin components. Besides providing information on the biological significance of these types of uv-induced lesions, this technique may be useful as a probe of chromatin structure

  16. Log-stable concentration distributions of trace elements in biomedical samples

    International Nuclear Information System (INIS)

    Kubala-Kukus, A.; Kuternoga, E.; Braziewicz, J.; Pajek, M.

    2004-01-01

    In the present paper, which follows our earlier observation that the asymmetric and long-tailed concentration distributions of trace elements in biomedical samples, measured by the X-ray fluorescence techniques, can be modeled by the log-stable distributions, further specific aspects of this observation are discussed. First, we demonstrate that, typically, for a quite substantial fraction (10-20%) of trace elements studied in different kinds of biomedical samples, the measured concentration distributions are described in fact by the 'symmetric' log-stable distributions, i.e. the asymmetric distributions which are described by the symmetric stable distributions. This observation is, in fact, expected for the random multiplicative process, which models the concentration distributions of trace elements in the biomedical samples. The log-stable nature of concentration distribution of trace elements results in several problems of statistical nature, which have to be addressed in XRF data analysis practice. Consequently, in the present paper, the following problems, namely (i) the estimation of parameters for stable distributions and (ii) the testing of the log-stable nature of the concentration distribution by using the Anderson-Darling (A 2 ) test, especially for symmetric stable distributions, are discussed in detail. In particular, the maximum likelihood estimation and Monte Carlo simulation techniques were used, respectively, for estimation of stable distribution parameters and calculation of the critical values for the Anderson-Darling test. The discussed ideas are exemplified by the results of the study of trace element concentration distributions in selected biomedical samples, which were obtained by using the X-ray fluorescence (XRF, TXRF) methods

  17. Concentrations of radiocesium and stable elements in different parts of pine tree collected in Chernobyl area

    International Nuclear Information System (INIS)

    Yoshida, Satoshi; Watanabe, Masumi; Suzuki, Akira; Linkov, Igor; Dvornik, Alexander; Zhuchenko, Tatiana

    2007-01-01

    Radial distributions of 137 Cs and related stable elements in a pine tree collected in Chernobyl contaminated area in Belarus were determined, in order to get basic information for dose estimation of pine tree. The concentration of 137 Cs in annual tree rings was the highest in cambium, and decreased sharply toward inside. The highest concentration of 137 Cs in cambium suggests the highest radiation dose to growing part of wood. Distribution of stable Cs was similar as that of 137 Cs, and the 137 Cs/stable Cs ratio was almost constant, indicating the equilibrium of Chernobyl 137 Cs with stable Cs in the pine wood. The similar distributions as Cs were observed for K and Rb. (author)

  18. Caffeine Citrate Dosing Adjustments to Assure Stable Caffeine Concentrations in Preterm Neonates.

    Science.gov (United States)

    Koch, Gilbert; Datta, Alexandre N; Jost, Kerstin; Schulzke, Sven M; van den Anker, John; Pfister, Marc

    2017-12-01

    To identify dosing strategies that will assure stable caffeine concentrations in preterm neonates despite changing caffeine clearance during the first 8 weeks of life. A 3-step simulation approach was used to compute caffeine doses that would achieve stable caffeine concentrations in the first 8 weeks after birth: (1) a mathematical weight change model was developed based on published weight distribution data; (2) a pharmacokinetic model was developed based on published models that accounts for individual body weight, postnatal, and gestational age on caffeine clearance and volume of distribution; and (3) caffeine concentrations were simulated for different dosing regimens. A standard dosing regimen of caffeine citrate (using a 20 mg/kg loading dose and 5 mg/kg/day maintenance dose) is associated with a maximal trough caffeine concentration of 15 mg/L after 1 week of treatment. However, trough concentrations subsequently exhibit a clinically relevant decrease because of increasing clearance. Model-based simulations indicate that an adjusted maintenance dose of 6 mg/kg/day in the second week, 7 mg/kg/day in the third to fourth week and 8 mg/kg/day in the fifth to eighth week assures stable caffeine concentrations with a target trough concentration of 15 mg/L. To assure stable caffeine concentrations during the first 8 weeks of life, the caffeine citrate maintenance dose needs to be increased by 1 mg/kg every 1-2 weeks. These simple adjustments are expected to maintain exposure to stable caffeine concentrations throughout this important developmental period and might enhance both the short- and long-term beneficial effects of caffeine treatment. Copyright © 2017 Elsevier Inc. All rights reserved.

  19. Thermal green protein, an extremely stable, nonaggregating fluorescent protein created by structure-guided surface engineering.

    Science.gov (United States)

    Close, Devin W; Paul, Craig Don; Langan, Patricia S; Wilce, Matthew C J; Traore, Daouda A K; Halfmann, Randal; Rocha, Reginaldo C; Waldo, Geoffery S; Payne, Riley J; Rucker, Joseph B; Prescott, Mark; Bradbury, Andrew R M

    2015-07-01

    In this article, we describe the engineering and X-ray crystal structure of Thermal Green Protein (TGP), an extremely stable, highly soluble, non-aggregating green fluorescent protein. TGP is a soluble variant of the fluorescent protein eCGP123, which despite being highly stable, has proven to be aggregation-prone. The X-ray crystal structure of eCGP123, also determined within the context of this paper, was used to carry out rational surface engineering to improve its solubility, leading to TGP. The approach involved simultaneously eliminating crystal lattice contacts while increasing the overall negative charge of the protein. Despite intentional disruption of lattice contacts and introduction of high entropy glutamate side chains, TGP crystallized readily in a number of different conditions and the X-ray crystal structure of TGP was determined to 1.9 Å resolution. The structural reasons for the enhanced stability of TGP and eCGP123 are discussed. We demonstrate the utility of using TGP as a fusion partner in various assays and significantly, in amyloid assays in which the standard fluorescent protein, EGFP, is undesirable because of aberrant oligomerization. © 2014 Wiley Periodicals, Inc.

  20. Levels of acute phase proteins remain stable after ischemic stroke

    Directory of Open Access Journals (Sweden)

    Paik Myunghee C

    2006-10-01

    Full Text Available Abstract Background Inflammation and inflammatory biomarkers play an important role in atherosclerosis and cardiovascular disease. Little information is available, however, on time course of serum markers of inflammation after stroke. Methods First ischemic stroke patients ≥40 years old had levels of high-sensitivity C-reactive protein (hsCRP, serum amyloid A (SAA, and fibrinogen measured in plasma samples drawn at 1, 2, 3, 7, 14, 21 and 28 days after stroke. Levels were log-transformed as needed, and parametric and non-parametric statistical tests were used to test for evidence of a trend in levels over time. Levels of hsCRP and SAA were also compared with levels in a comparable population of stroke-free participants. Results Mean age of participants with repeated measures (n = 21 was 65.6 ± 11.6 years, and 13 (61.9% were men, and 15 (71.4% were Hispanic. Approximately 75% of patients (n = 15 had mild strokes (NIH Stroke Scale score 0–5. There was no evidence of a time trend in levels of hsCRP, SAA, or fibrinogen for any of the markers during the 28 days of follow-up. Mean log(hsCRP was 1.67 ± 1.07 mg/L (median hsCRP 6.48 mg/L among stroke participants and 1.00 ± 1.18 mg/L (median 2.82 mg/L in a group of 1176 randomly selected stroke-free participants from the same community (p = 0.0252. Conclusion Levels of hsCRP are higher in stroke patients than in stroke-free subjects. Levels of inflammatory biomarkers associated with atherosclerosis, including hsCRP, appear to be stable for at least 28 days after first ischemic stroke.

  1. Highly stable, protein capped gold nanoparticles as effective drug delivery vehicles for amino-glycosidic antibiotics

    International Nuclear Information System (INIS)

    Rastogi, Lori; Kora, Aruna Jyothi; Arunachalam, J.

    2012-01-01

    A method for the production of highly stable gold nanoparticles (Au NP) was optimized using sodium borohydride as reducing agent and bovine serum albumin as capping agent. The synthesized nanoparticles were characterized using UV–visible spectroscopy, transmission electron microscopy, X‐ray diffraction (XRD) and dynamic light scattering techniques. The formation of gold nanoparticles was confirmed from the appearance of pink colour and an absorption maximum at 532 nm. These protein capped nanoparticles exhibited excellent stability towards pH modification and electrolyte addition. The produced nanoparticles were found to be spherical in shape, nearly monodispersed and with an average particle size of 7.8 ± 1.7 nm. Crystalline nature of the nanoparticles in face centered cubic structure is confirmed from the selected‐area electron diffraction and XRD patterns. The nanoparticles were functionalized with various amino-glycosidic antibiotics for utilizing them as drug delivery vehicles. Using Fourier transform infrared spectroscopy, the possible functional groups of antibiotics bound to the nanoparticle surface have been examined. These drug loaded nanoparticle solutions were tested for their antibacterial activity against Gram-negative and Gram-positive bacterial strains, by well diffusion assay. The antibiotic conjugated Au NP exhibited enhanced antibacterial activity, compared to pure antibiotic at the same concentration. Being protein capped and highly stable, these gold nanoparticles can act as effective carriers for drugs and might have considerable applications in the field of infection prevention and therapeutics. - Highlights: ► Method for NaBH 4 reduced and BSA capped gold nanoparticle was standardized. ► Nanoparticles were spherical and nearly monodispersed with a size of 7.8 nm. ► Nanoparticles are extremely stable towards pH modification and electrolyte addition. ► Antibiotic conjugated nanoparticles exhibited enhanced antibacterial activity

  2. Mis-translation of a Computationally Designed Protein Yields an Exceptionally Stable Homodimer: Implications for Protein Engineering and Evolution.

    Energy Technology Data Exchange (ETDEWEB)

    Dantas, Gautam; Watters, Alexander L.; Lunde, Bradley; Eletr, Ziad; Isern, Nancy G.; Roseman, Toby; Lipfert, Jan; Doniach, Sebastian; Tompa, Martin; Kuhlman, Brian; Stoddard, Barry L.; Varani, Gabriele; Baker, David

    2006-10-06

    We recently used computational protein design to create an extremely stable, globular protein, Top7, with a sequence and fold not observed previously in nature. Since Top7 was created in the absence of genetic selection, it provides a rare opportunity to investigate aspects of the cellular protein production and surveillance machinery that are subject to natural selection. Here we show that a portion of the Top7 protein corresponding to the final 49 C-terminal residues is efficiently mistranslated and accumulates at high levels in E. coli. We used circular dichroism spectroscopy, size-exclusion chromatography, small-angle x-ray scattering, analytical ultra-centrifugation, and NMR spectroscopy to show that the resulting CFr protein adopts a compact, extremely-stable, obligate, symmetric, homo-dimeric structure. Based on the solution structure, we engineered an even more stable variant of CFr by disulfide-induced covalent circularisation that should be an excellent platform for design of novel functions. The accumulation of high levels of CFr exposes the high error rate of the protein translation machinery, and the rarity of correspondingly stable fragments in natural proteins implies a stringent evolutionary pressure against protein sub-fragments that can independently fold into stable structures. The symmetric self-association between two identical mistranslated CFr sub-units to generate an extremely stable structure parallels a mechanism for natural protein-fold evolution by modular recombination of stable protein sub-structures.

  3. Experimental investigation of concentration and stable isotopes signals during organic contaminants back diffusion

    DEFF Research Database (Denmark)

    Jin, Biao; Nika, Chrysanthi-Elisabeth; Rolle, Massimo

    2017-01-01

    -dichloroethene (cis-DCE) as model contaminant and we investigated its back diffusion from an impermeable source into a permeable saturated layer, in which advection-dominated flow conditions were established. We used concentration and stable chlorine isotope measurements to investigate the plumes originated by cis...... and stable isotope gradients in the flow-through setup. In particular, steep concentration and stable isotope gradients were observed at the outlet. Lateral isotope gradients corresponding to chlorine isotope fractionation up to 20‰ were induced by cis-DCE back diffusion and subsequent advection......-dominated transport in all flow-through experiments. A numerical modeling approach, tracking individually all chlorine isotopologues, based on the accurate parameterization of local dispersion, as well as on the values of aqueous diffusion coefficients and diffusion-induced isotope fractionation from a previous study...

  4. The effects of dietary energy and protein concentrations on ostrich ...

    African Journals Online (AJOL)

    The effects were investigated of energy and protein concentrations (with associated amino acid concentrations) in ostrich diets on leather quality of the skins of 50 ostriches. Energy concentrations were 9.0, 10.5 and 12.0 MJ ME/kg diet and protein concentrations were 130, 150 and 170 g/kg diet. The physical leather ...

  5. Determining synthesis rates of individual proteins in zebrafish (Danio rerio) with low levels of a stable isotope labelled amino acid.

    Science.gov (United States)

    Geary, Bethany; Magee, Kieran; Cash, Phillip; Young, Iain S; Whitfield, Phillip D; Doherty, Mary K

    2016-05-01

    The zebrafish is a powerful model organism for the analysis of human cardiovascular development and disease. Understanding these processes at the protein level not only requires changes in protein concentration to be determined but also the rate at which these changes occur on a protein-by-protein basis. The ability to measure protein synthesis and degradation rates on a proteome-wide scale, using stable isotope labelling in conjunction with mass spectrometry is now a well-established experimental approach. With the advent of more selective and sensitive mass spectrometers, it is possible to accurately measure lower levels of stable isotope incorporation, even when sample is limited. In order to challenge the sensitivity of this approach, we successfully determined the synthesis rates of over 600 proteins from the cardiac muscle of the zebrafish using a diet where either 30% or 50% of the L-leucine was replaced with a stable isotope labelled analogue ([(2) H7 ]L-leucine]. It was possible to extract sufficient protein from individual zebrafish hearts to determine the incorporation rate of the label into hundreds of proteins simultaneously, with the two labelling regimens showing a good correlation of synthesis rates. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Refractometric total protein concentrations in icteric serum from dogs.

    Science.gov (United States)

    Gupta, Aradhana; Stockham, Steven L

    2014-01-01

    To determine whether high serum bilirubin concentrations interfere with the measurement of serum total protein concentration by refractometry and to assess potential biases among refractometer measurements. Evaluation study. Sera from 2 healthy Greyhounds. Bilirubin was dissolved in 0.1M NaOH, and the resulting solution was mixed with sera from 2 dogs from which food had been withheld to achieve various bilirubin concentrations up to 40 mg/dL. Refractometric total protein concentrations were estimated with 3 clinical refractometers. A biochemical analyzer was used to measure biuret assay-based total protein and bilirubin concentrations with spectrophotometric assays. No interference with refractometric measurement of total protein concentrations was detected with bilirubin concentrations up to 41.5 mg/dL. Biases in refractometric total protein concentrations were detected and were related to the conversion of refractive index values to total protein concentrations. Hyperbilirubinemia did not interfere with the refractometric estimation of serum total protein concentration. The agreement among total protein concentrations estimated by 3 refractometers was dependent on the method of conversion of refractive index to total protein concentration and was independent of hyperbilirubinemia.

  7. Identification and characterization of stable membrane protein complexes

    NARCIS (Netherlands)

    Spelbrink, R.E.J.

    2007-01-01

    Many membrane proteins exist as oligomers. Such oligomers play an important role in a broad variety of cellular processes such as ion transport, energy transduction, osmosensing and cell wall synthesis. We developed an electrophoresis-based method of identifying oligomeric membrane proteins that are

  8. Production of protein concentrate and isolate from cashew ...

    African Journals Online (AJOL)

    The protein isolates were obtained by an alkaline extraction-isoelectric precipitation method, which involved aqueous alkaline extraction of the proteins at low temperature, and isoelectric precipitation of the protein fractions; the protein concentrates were obtained using an alkaline extraction-methanol precipitation method, ...

  9. Heat-stable proteins and abscisic acid action in barley aleurone cells

    International Nuclear Information System (INIS)

    Jacobsen, J.V.; Shaw, D.C.

    1989-01-01

    [ 35 S]Methionine labeling experiments showed that abscisic acid (ABA) induced the synthesis of at least 25 polypeptides in mature barley (Hordeum vulgare) aleurone cells. The polypeptides were not secreted. Whereas most of the proteins extracted from aleurone cells were coagulated by heating to 100 degree C for 10 minutes, most of the ABA-induced polypeptides remained in solution (heat-stable). ABA had little effect on the spectrum of polypeptides that were synthesized and secreted by aleurone cells, and most of these secreted polypeptides were also heat-stable. Coomassie blue staining of sodium dodecyl sulfate polyacrylamide gels indicated that ABA-induced polypeptides already occurred in high amounts in mature aleurone layers having accumulated during grain development. About 60% of the total protein extracted from mature aleurone was heat stable. Amino acid analyses of total preparations of heat-stable and heat-labile proteins showed that, compared to heat-labile proteins, heat-stable intracellular proteins were characterized by higher glutamic acid/glutamine (Glx) and glycine levels and lower levels of neutral amino acids. Secreted heat-stable proteins were rich in Glx and proline. The possibilities that the accumulation of the heat-stable polypeptides during grain development is controlled by ABA and that the function of these polypeptides is related to their abundance and extraordinary heat stability are considered

  10. Nutritional and functional properties of whey proteins concentrate and isolate

    Directory of Open Access Journals (Sweden)

    Zoran Herceg

    2006-12-01

    Full Text Available Whey protein fractions represent 18 - 20 % of total milk nitrogen content. Nutritional value in addition to diverse physico - chemical and functional properties make whey proteins highly suitable for application in foodstuffs. In the most cases, whey proteins are used because of their functional properties. Whey proteins possess favourable functional characteristics such as gelling, water binding, emulsification and foaming ability. Due to application of new process techniques (membrane fractionation techniques, it is possible to produce various whey - protein based products. The most important products based on the whey proteins are whey protein concentrates (WPC and whey protein isolates (WPI. The aim of this paper was to give comprehensive review of nutritional and functional properties of the most common used whey proteins (whey protein concentrate - WPC and whey protein isolate - WPI in the food industry.

  11. Studies on concentration of minor stable elements in marine environmental samples

    International Nuclear Information System (INIS)

    Suzuki, Hamaji; Ishii, Toshiaki; Iimura, Mitsue; Koyanagi, Taku

    1978-01-01

    Information on the physico-chemical state and quantity of stable elements in marine environments is frequently required to analyze the radioecological behavior of radionuclides released from nuclear facilities into the sea. In this work, determination of stable Mn, Fe, Co, Zn, Zr, Rb, Cs and some rare earth elements (Ce, Eu, Tb, Yb and Lu) in seawater and marine organisms was carried out and the concentration factors were estimated. Seawater and marine organisms were collected on the seashore of Ibaraki Prefecture and analysed by means of neutron activation analysis or atomic absorption spectrometry depending on the elements or samples. Average concentration factors of the rare earth elements by marine organisms are estimated as 3 x 10 1 : muscle of fish, 5 x 10 2 : soft part of clams, and 2 x 10 2 : algae, respectively. Concentration factors by muscle of fishes were 10 3 : Fe, 2 x 10 2 : Co, 5 x 10 2 : Zn, and 5 x 10 1 : Cs, and those by soft part of shellfishes were 10 4 : Fe, 10 3 : Co, 2 x 10 3 : Zn, and 10 1 : Cs, whereas those by algae were 2 x 10 4 : Fe, 5 x 10 2 : Co, 10 3 : Zn, and 3 x 10 1 : Cs, respectively. The high concentration factors for numerous stable elements by shellfishes and algae suggested their suitabilities to the indicator organisms for monitoring of marine pollution by these heavy metals and corresponding radioisotopes and also their significant contribution to the internal radiation exposure to man as radioactive seafoods. (author)

  12. Urine protein concentration estimation for biomarker discovery

    OpenAIRE

    Mistry, Hiten D.; Bramham, Kate; Weston, Andrew; Ward, Malcolm; Thompson, Andrew; Chappell, Lucy C.

    2013-01-01

    Recent advances have been made in the study of urinary proteomics as a diagnostic tool for renal disease and pre-eclampsia which requires accurate measurement of urinary protein. We compared different protein assays (Bicinchoninic acid (BCA), Lowry and Bradford) against the ‘gold standard’ amino-acid assay in urine from 43 women (8 non-pregnant, 34 pregnant, including 8 with pre-eclampsia. BCA assay was superior to both Lowry and Bradford assays (Bland Altman bias: 0.08) compared to amino-aci...

  13. Extremely stable soluble high molecular mass multi-protein complex with DNase activity in human placental tissue.

    Directory of Open Access Journals (Sweden)

    Evgeniya E Burkova

    Full Text Available Human placenta is an organ which protects, feeds, and regulates the grooving of the embryo. Therefore, identification and characterization of placental components including proteins and their multi-protein complexes is an important step to understanding the placenta function. We have obtained and analyzed for the first time an extremely stable multi-protein complex (SPC, ∼ 1000 kDa from the soluble fraction of three human placentas. By gel filtration on Sepharose-4B, the SPC was well separated from other proteins of the placenta extract. Light scattering measurements and gel filtration showed that the SPC is stable in the presence of NaCl, MgCl2, acetonitrile, guanidinium chloride, and Triton in high concentrations, but dissociates efficiently in the presence of 8 M urea, 50 mM EDTA, and 0.5 M NaCl. Such a stable complex is unlikely to be a casual associate of different proteins. According to SDS-PAGE and MALDI mass spectrometry data, this complex contains many major glycosylated proteins with low and moderate molecular masses (MMs 4-14 kDa and several moderately abundant (79.3, 68.5, 52.8, and 27.2 kDa as well as minor proteins with higher MMs. The SPC treatment with dithiothreitol led to a disappearance of some protein bands and revealed proteins with lower MMs. The SPCs from three placentas efficiently hydrolyzed plasmid supercoiled DNA with comparable rates and possess at least two DNA-binding sites with different affinities for a 12-mer oligonucleotide. Progress in study of placental protein complexes can promote understanding of their biological functions.

  14. 21 CFR 172.385 - Whole fish protein concentrate.

    Science.gov (United States)

    2010-04-01

    ... CONSUMPTION Special Dietary and Nutritional Additives § 172.385 Whole fish protein concentrate. The food additive whole fish protein concentrate may be safely used as a food supplement in accordance with the... fish that are used in other forms for human food. (b) The additive consists essentially of a dried fish...

  15. Total protein and cholesterol concentrations in brain regions of male ...

    African Journals Online (AJOL)

    The results showed similarities (P>0.05) between the treatments in total protein concentrations in the cerebral cortex, medulla, hypothalamus, amygdala, mesencephalon and hippocampus. Total protein concentrations however differed significantly between diets (P<0.05) in the cerebellum and pons varoli with the lowest ...

  16. Highly Stable Trypsin-Aggregate Coatings on Polymer Nanofibers for Repeated Protein Digestion

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Byoung Chan; Lopez-Ferrer, Daniel; Lee, Sang-mok; Ahn, Hye-kyung; Nair, Sujith; Kim, Seong H.; Kim, Beom S.; Petritis, Konstantinos; Camp, David G.; Grate, Jay W.; Smith, Richard D.; Koo, Yoon-mo; Gu, Man Bock; Kim, Jungbae

    2009-04-01

    A stable and robust trypsin-based biocatalytic system was developed and demonstrated for proteomic applications. The system utilizes polymer nanofibers coated with trypsin aggregates for immobilized protease digestions. After covalently attaching an initial layer of trypsin to the polymer nanofibers, highly concentrated trypsin molecules are crosslinked to the layered trypsin by way of a glutaraldehyde treatment. This new process produced a 300-fold increase in trypsin activity compared with a conventional method for covalent trypsin immobilization and proved to be robust in that it still maintained a high level of activity after a year of repeated recycling. This highly stable form of immobilized trypsin was also resistant to autolysis, enabling repeated digestions of bovine serum albumin over 40 days and successful peptide identification by LC-MS/MS. Finally, the immobilized trypsin was resistant to proteolysis when exposed to other enzymes (i.e. chymotrypsin), which makes it suitable for use in “real-world” proteomic applications. Overall, the biocatalytic nanofibers with enzyme aggregate coatings proved to be an effective approach for repeated and automated protein digestion in proteomic analyses.

  17. Radioactive and stable cobalt concentrations in mussel in Kyushu island, Japan

    International Nuclear Information System (INIS)

    Momoshima, Noriyuki; Shiki, Atsushi; Takashima, Yoshimasa; Maki, Takao; Koriyama, Munehiro; Shimozono, Seika; Imamura, Hiroka; Nakamata, Kojiro.

    1985-01-01

    Two kinds of mussel, Septifer virgatus and Mytilus edulis, were collected from Kyushu island, Japan, in order to elucidate a background level of 60 Co, which is one of the most significant radionuclide for environmental monitoring around a nuclear power plant. The mussels were collected from 7 locations in 1983 and classified 2 or 3 groups depending on their shell size at each location. Activities of 60 Co were measured by a low-background β counter after purified by means of chemical separation and electrodeposition. Stable cobalt concentrations were determined by colorimetric method. The concentrations of cobalt in Septifer virgatus are one order higher level than that in Mytilus edulis. There are not so large difference in cobalt content depending on shell size so long as comparing them at the same location. The radioactivities in mussels show the same trend as stable cobalt. It has become apparent that Septifer virgatus has a tendency to concentrate cobalt with growing but Mytilus edulis is opposite. The cobalt-60 introduced to sea from nuclear explosions seems to be relatively constant in coastal seawater since specific activities are distributed in a narrow range in spite of kind, shell size and location. (author)

  18. Concentration factors of stable elements and radionuclides in Po river fish

    International Nuclear Information System (INIS)

    Achilli, M.; Ciceri, G.; Bozzani, A.; Guzzi, L.; Queirazza, G.

    1988-01-01

    The concentration factors (CF) of stable Co, Cs, Mn, Fe, Zn and Sr in different fish from six stretches in the middle course of the Po river (N. Italy) have been investigated. The space-time variation in water has been followed for 14 months. The investigation has been undertaken to study CF variations in the same fish species as a function of the physico-chemical form of the different elements in water (dissolved, dissolved and exchangeable fraction of the particulate, total). CF values of 103 Ru, 131 I and 134 - 137 Cs were also investigated for Cyprinus carpio reared, with artificial food, in two semi-natural environments

  19. Concentrations of radionuclides and selected stable elements in fruits and vegetables

    International Nuclear Information System (INIS)

    Oakes, T.W.; Shank, K.E.

    1977-01-01

    Twenty-two types of fruits and vegetables collected from two commercial supermarkets have been analyzed for their radionuclidic and stable-element composition. A specific gamma-emitting isotope analysis was performed on each sample for 40 K, 60 Co, 95 Zr-Nb, 106 Ru, 137 Cs, 226 Ra, and 232 Th. The concentration of the stable elements in each sample were determined using multi-element neutron-activation analysis (Al, Ag, Au, As, Ba, Br, Ca, Ce, Cl, Co, Cr, Cs, Cu, Eu, Fe, Hf, I, K, La, Mn, Mo, Mg, Na, Rb, Sb, Sc, Se, Sr, Ta, Te, Th, Ti, Zn, Zr) and atomic absorption (Cd, Ni, Pb). Information on the composition of a typical diet is used to estimate the radiological dose to man subsequent to ingestion of these fruits and vegetables. The stable-elemental compositions of the foodstuffs analyzed were compared with estimated values assuming foliar deposition and long-term buildup of effluents from a large modern coal-fired steam plant. It is tentatively concluded that for the general case of a precipitator-equipped, coal-fired steam plant, no toxic levels of trace elements in foodstuffs are expected as a result of the plant operation

  20. Influence of Bleaching on Flavor of 34% Whey Protein Concentrate and Residual Benzoic Acid Concentration in Dried Whey Proteins

    Science.gov (United States)

    Previous studies have shown that bleaching negatively affects the flavor of 70% whey protein concentrate (WPC70), but bleaching effects on lower-protein products have not been established. Benzoyl peroxide (BP), a whey bleaching agent, degrades to benzoic acid (BA) and may elevate BA concentrations...

  1. TGP, an extremely stable, non-aggregating fluorescent protein created by structure-guided surface engineering

    OpenAIRE

    Close, Devin W.; Don Paul, Craig; Langan, Patricia S.; Wilce, Matthew C.J.; Traore, Daouda A.K.; Halfmann, Randal; Rocha, Reginaldo C.; Waldo, Geoffery S.; Payne, Riley J.; Rucker, Joseph B.; Prescott, Mark; Bradbury, Andrew R.M.

    2015-01-01

    In this paper we describe the engineering and X-ray crystal structure of Thermal Green Protein (TGP), an extremely stable, highly soluble, non-aggregating green fluorescent protein. TGP is a soluble variant of the fluorescent protein eCGP123, which despite being highly stable, has proven to be aggregation-prone. The X-ray crystal structure of eCGP123, also determined within the context of this paper, was used to carry out rational surface engineering to improve its solubility, leading to TGP....

  2. Insights from quantitative metaproteomics and protein-stable isotope probing into microbial ecology.

    Science.gov (United States)

    von Bergen, Martin; Jehmlich, Nico; Taubert, Martin; Vogt, Carsten; Bastida, Felipe; Herbst, Florian-Alexander; Schmidt, Frank; Richnow, Hans-Hermann; Seifert, Jana

    2013-10-01

    The recent development of metaproteomics has enabled the direct identification and quantification of expressed proteins from microbial communities in situ, without the need for microbial enrichment. This became possible by (1) significant increases in quality and quantity of metagenome data and by improvements of (2) accuracy and (3) sensitivity of modern mass spectrometers (MS). The identification of physiologically relevant enzymes can help to understand the role of specific species within a community or an ecological niche. Beside identification, relative and absolute quantitation is also crucial. We will review label-free and label-based methods of quantitation in MS-based proteome analysis and the contribution of quantitative proteome data to microbial ecology. Additionally, approaches of protein-based stable isotope probing (protein-SIP) for deciphering community structures are reviewed. Information on the species-specific metabolic activity can be obtained when substrates or nutrients are labeled with stable isotopes in a protein-SIP approach. The stable isotopes ((13)C, (15)N, (36)S) are incorporated into proteins and the rate of incorporation can be used for assessing the metabolic activity of the corresponding species. We will focus on the relevance of the metabolic and phylogenetic information retrieved with protein-SIP studies and for detecting and quantifying the carbon flux within microbial consortia. Furthermore, the combination of protein-SIP with established tools in microbial ecology such as other stable isotope probing techniques are discussed.

  3. Concentrations of stable elements and uranium in estuarine areas of Japan

    International Nuclear Information System (INIS)

    Takata, Hyoe; Aono, Tatsuo; Tagami, Keiko; Uchida, Shigeo

    2008-01-01

    The geochemistry of stable elements can be a good analogue for understanding the behavior of radionuclides in estuarine and coastal environments. In this study, the behavior of nutrients (NO 3 + NO 2 , PO 4 , Si(OH) 4 ), heavy metals, and U was observed in several estuarine and coastal waters of Japan. We also collected data on salinity, pH, and suspended particle matter (SPM). Nutrient concentrations followed conservative dilution lines in these estuaries, and concentrations of dissolved Fe decreased as salinity increased from 0 to 20. In general, most of the dissolved Fe in estuaries is in colloidal form. The behavior of dissolved Fe might reflect a loss of colloidal Fe through coagulation in this salinity range. Dissolved Co and Ni concentrations followed approximate dilution lines from the rivers to the seawater end-members, suggesting that they were quasi-conservative in these estuarine systems. A rapid increase in dissolved Cd concentrations was observed at low levels of salinity (<4). Estimated fluxes of dissolved Cd to the estuarine and coastal waters showed that the salt-induced desorption of Cd from particles constitutes a significant source of dissolved Cd in the estuarine and coastal waters. (author)

  4. Quantification of protein concentration using UV absorbance and Coomassie dyes.

    Science.gov (United States)

    Noble, James E

    2014-01-01

    The measurement of a solubilized protein concentration in solution is an important assay in biochemistry research and development labs for applications ranging from enzymatic studies to providing data for biopharmaceutical lot release. Spectrophotometric protein quantification assays are methods that use UV and visible spectroscopy to rapidly determine the concentration of protein, relative to a standard, or using an assigned extinction coefficient. Where multiple samples need measurement, and/or the sample volume and concentration is limited, preparations of the Coomassie dye commonly known as the Bradford assay can be used. © 2014 Elsevier Inc. All rights reserved.

  5. Identification of a lysosome membrane protein which could mediate ATP-dependent stable association of lysosomes to microtubules

    International Nuclear Information System (INIS)

    Mithieux, G.; Rousset, B.

    1989-01-01

    We have previously reported that purified thyroid lysosomes bind to reconstituted microtubules to form stable complexes, a process which is inhibited by ATP. Among detergent-solubilized lysosomal membrane protein, we identified a 50-kDa molecular component which binds to preassembled microtubules. The binding of this polypeptide to microtubules was decreased in the presence of ATP. We purified this 50-kDa protein by affinity chromatography on immobilized ATP. The 50-kDa protein bound to the ATP column was eluted by 1 mM ATP. The purified protein, labeled with 125I, exhibited the ability of interacting with microtubules. The binding process was inhibited by increasing concentrations of ATP, the half-maximal inhibitory effect being obtained at an ATP concentration of 0.35 mM. The interaction of the 50-kDa protein with microtubules is a saturable phenomenon since the binding of the 125I-labeled 50-kDa protein was inhibited by unlabeled solubilized lysosomal membrane protein containing the 50-kDa polypeptide but not by the same protein fraction from which the 50-kDa polypeptide had been removed by the ATP affinity chromatography procedure. The 50-kDa protein has the property to bind to pure tubulin coupled to an insoluble matrix. The 50-kDa protein was eluted from the tubulin affinity column by ATP. These findings support the conclusion that a protein inserted into the lysosomal membrane is able to bind directly to microtubules in a process which can be regulated by ATP. We propose that this protein could account for the association of lysosomes to microtubules demonstrated both in vitro and in intact cells

  6. Comparison of total protein concentration in skeletal muscle as measured by the Bradford and Lowry assays.

    Science.gov (United States)

    Seevaratnam, Rajini; Patel, Barkha P; Hamadeh, Mazen J

    2009-06-01

    The Lowry and Bradford assays are the most commonly used methods of total protein quantification, yet vary in several aspects. To date, no comparisons have been made in skeletal muscle. We compared total protein concentrations of mouse red and white gastrocnemius, reagent stability, protein stability and range of linearity using both assays. The Lowry averaged protein concentrations 15% higher than the Bradford with a moderate correlation (r = 0.36, P = 0.01). However, Bland-Altman analysis revealed considerable bias (15.8 +/- 29.7%). Both Lowry reagents and its protein-reagent interactions were less stable over time than the Bradford. The linear range of concentration was smaller for the Lowry (0.05-0.50 mg/ml) than the Bradford (0-2.0 mg/ml). We conclude that the Bradford and Lowry measures of total protein concentration in skeletal muscle are not interchangeable. The Bradford and Lowry assays have various strengths and weaknesses in terms of substance interference and protein size. However, the Bradford provides greater reagent stability, protein-reagent stability and range of linearity, and requires less time to analyse compared to the Lowry assay.

  7. β-trace protein (prostaglandin D synthase - a stable and reliable protein in perilymph

    Directory of Open Access Journals (Sweden)

    Nekic, Marko

    2005-06-01

    Full Text Available Objective: Beta-trace protein (β-TP has been analysed in human cerebrospinal fluid (CSF and other body fluids. Beta-trace protein is a very sensitive and specific clinical marker and can confirm reliably the presence of CSF in patients with a suspected CSF leakage. Design: Perilymph specimens from the scala vestibuli (n=10 and from the lateral semicircular canal (n=4 were taken from patients undergoing stapedotomy or surgery for acoustic neuroma. During post-mortem examinations perilymph specimens from the scala vestibuli (n=70, the scala tympani (n=11, endolymph specimens (n=21 and CSF specimens (n=17 were obtained. All specimens were analyzed by a one-dimensional immunoelectrophoresis using a polyclonal, monospecific antibody. Results: Specimens from live surgery showed a mean concentration of 51.5 ± 48.9 mg/l β-TP in scala vestibuli perilymph. Specimens from post-mortem examinations revealed a mean concentration of 49.1 ± 17.7 mg/l in CSF, 71.9 ± 29.3 mg/l in perilymph and 68.0 ± 21.7 mg/l in endolymph. There was no evidence of a circadian alteration of β-TP in CSF or inner ear fluids. Conclusions: Our results demonstrated clearly that β-TP is contained in human perilymph and endolymph. This is the first published data that point out the aptitude of the β-TP-test in verifying traces of perilymph, a valuable diagnostic tool for the existence of perilymphatic leaks.

  8. Inactive Doses and Protein Concentration of Gamma Irradiated Yersinia Enterocolitica

    International Nuclear Information System (INIS)

    Irawan Sugoro; Sandra Hermanto

    2009-01-01

    Yersinia enterocolitica is one of bacteria which cause coliform mastitis in dairy cows. The bacteria could be inactivated by gamma irradiation as inactivated vaccine candidate. The experiment has been conducted to determine the inactive doses and the protein concentration of Yersinia enterocolitica Y3 which has been irradiated by gamma rays. The cells cultures were irradiated by gamma rays with doses of 0, 100, 200, 400, 600, 800, 1.000 and 1.500 Gy (doses rate was 1089,59 Gy/hours). The inactive dose was determined by the drop test method and the protein concentration of cells were determined by Lowry method. The results showed that the inactive doses occurred on 800 – 1500 Gy. The different irradiation doses of cell cultures showed the effect of gamma irradiation on the protein concentration that was random and has a significant effect on the protein concentration. (author)

  9. Effect of Limited Hydrolysis on Traditional Soy Protein Concentrate

    Directory of Open Access Journals (Sweden)

    Mirjana B. Pesic

    2006-09-01

    Full Text Available The influence of limited proteolysis of soy protein concentrate on proteinextractability, the composition of the extractable proteins, their emulsifying properties andsome nutritional properties were investigated. Traditional concentrate (alcohol leachedconcentrate was hydrolyzed using trypsin and pepsin as hydrolytic agents. Significantdifferences in extractable protein composition between traditional concentrate and theirhydrolysates were observed by polyacrylamide gel electrophoresis (PAGE and by SDSPAGE.All hydrolysates showed better extractability than the original protein concentrate,whereas significantly better emulsifying properties were noticed at modified concentratesobtained by trypsin induced hydrolysis. These improved properties are the result of twosimultaneous processes, dissociation and degradation of insoluble alcohol-induced proteinaggregates. Enzyme induced hydrolysis had no influence on trypsin-inibitor activity, andsignificantly reduced phytic acid content.

  10. Nutrient Concentrations and Stable Isotopes of Runoff from a Midwest Tile-Drained Corn Field

    Science.gov (United States)

    Wilkins, B. P.; Woo, D.; Li, J.; Michalski, G. M.; Kumar, P.; Conroy, J. L.; Keefer, D. A.; Keefer, L. L.; Hodson, T. O.

    2017-12-01

    Tile drains are a common crop drainage device used in Midwest agroecosystems. While efficient at drainage, the tiles provide a quick path for nutrient runoff, reducing the time available for microbes to use nutrients (e.g., NO3- and PO43-) and reduce export to riverine systems. Thus, understanding the effects of tile drains on nutrient runoff is critical to achieve nutrient reduction goals. Here we present isotopic and concentration data collected from tile drain runoff of a corn field located near Monticello, IL. Tile flow samples were measured for anion concentrations and stable isotopes of H2O and NO3-, while precipitation was measured for dual isotopes of H2O. Results demonstrate early tile flow from rain events have a low Cl- concentration (60% contribution) in the beginning of the hydrograph. As flow continues H2O isotopic values reflect pre-event water (ground and soil water), and Cl- concentrations increase representing a greater influence by matrix flow (60-90% contribution). Nitrate concentrations change dramatically, especially during the growing season, and do not follow a similar trend as the conservative Cl-, often decreasing days before, which represents missing nitrate in the upper surface portion of the soil. Nitrate isotopic data shows significant changes in 15N (4‰) and 18O (4‰) during individual hydrological events, representing that in addition to plant uptake and leaching, considerate NO3- is lost through denitrification. It is notable, that throughout the season d15N and d18O of nitrate change significantly representing that seasonally, substantial denitrification occurs.

  11. Effects of trace element concentration on enzyme controlled stable isotope fractionation during aerobic biodegradation of toluene.

    Science.gov (United States)

    Mancini, Silvia A; Hirschorn, Sarah K; Elsner, Martin; Lacrampe-Couloume, Georges; Sleep, Brent E; Edwards, Elizabeth A; Lollar, Barbara Sherwood

    2006-12-15

    The effects of iron concentration on carbon and hydrogen isotopic fractionation during aerobic biodegradation of toluene by Pseudomonas putida mt-2 were investigated using a low iron medium and two different high iron media. Mean carbon enrichment factors (epsilonc) determined using a Rayleigh isotopic model were smaller in culture grown under high iron conditions (epsilonc = -1.7+/-0.1%) compared to low iron conditions (epsilonc = -2.5+/-0.3%). Mean hydrogen enrichment factors (epsilonH) were also significantly smaller for culture grown under high iron conditions (epsilonH = -77 +/-4%) versus low iron conditions (EpsilonH = -159+/-11%). A mechanistic model for enzyme kinetics was used to relate differences in the magnitude of isotopic fractionation for low iron versus high iron cultures to the efficiency of the enzymatic transformation. The increase of carbon and hydrogen enrichment factors at low iron concentrations suggests a slower enzyme-catalyzed substrate conversion step (k2) relative to the enzyme-substrate binding step (k-l) at low iron concentration. While the observed differences were subtle and, hence, do not significantly impact the ability to use stable isotope analysis in the field, these results demonstrated that resolvable differences in carbon and hydrogen isotopic fractionation were related to low and high iron conditions. This novel result highlights the need to further investigate the effects of other trace elements known to be key components of biodegradative enzymes.

  12. Plasma Protein Turnover Rates in Rats Using Stable Isotope Labeling, Global Proteomics, and Activity-Based Protein Profiling

    Energy Technology Data Exchange (ETDEWEB)

    Smith, Jordan N.; Tyrrell, Kimberly J.; Hansen, Joshua R.; Thomas, Dennis G.; Murphree, Taylor A.; Shukla, Anil K.; Luders, Teresa; Madden, James M.; Li, Yunying; Wright, Aaron T.; Piehowski, Paul D.

    2017-12-06

    Protein turnover is important for general health on cellular and organism scales providing a strategy to replace old, damaged, or dysfunctional proteins. Protein turnover also informs of biomarker kinetics, as a better understanding of synthesis and degradation of proteins increases the clinical utility of biomarkers. Here, turnover rates of plasma proteins in rats were measured in vivo using a pulse-chase stable isotope labeling experiment. During the pulse, rats (n=5) were fed 13C6-labeled lysine (“heavy”) feed for 23 days to label proteins. During the chase, feed was changed to an unlabeled equivalent feed (“light”), and blood was repeatedly sampled from rats over 10 time points for 28 days. Plasma samples were digested with trypsin, and analyzed with liquid chromatography-tandem mass spectrometry (LC-MS/MS). MaxQuant was used to identify peptides and proteins, and quantify heavy:light lysine ratios. A system of ordinary differential equations was used to calculate protein turnover rates. Using this approach, 273 proteins were identified, and turnover rates were quantified for 157 plasma proteins with half-lives ranging 0.3-103 days. For the ~70 most abundant proteins, variability in turnover rates among rats was low (median coefficient of variation: 0.09). Activity-based protein profiling was applied to pooled plasma samples to enrich serine hydrolases using a fluorophosphonate (FP2) activity-based probe. This enrichment resulted in turnover rates for an additional 17 proteins. This study is the first to measure global plasma protein turnover rates in rats in vivo, measure variability of protein turnover rates in any animal model, and utilize activity-based protein profiling for enhancing measurements of targeted, low-abundant proteins, such as those commonly used as biomarkers. Measured protein turnover rates will be important for understanding of the role of protein turnover in cellular and organism health as well as increasing the utility of protein

  13. Behavior of whey protein concentrates under extreme storage conditions

    Science.gov (United States)

    The overseas demand for whey protein concentrates (WPC) has increased steadily in recent years. Emergency aid foods often include WPC, but shelf-life studies of whey proteins under different shipment and storage conditions have not been conducted in the last 50 yr. Microbial quality, compound form...

  14. Nutritive evaluation of Telfairia occidentalis leaf protein concentrate ...

    African Journals Online (AJOL)

    Leaf meal (LM), leaf proteins concentrate (LPC) and LPC residues from Telfairia occidentalis were produced, chemically characterized and the protein quality of the LPC evaluated using rats. Five infant weaning foods were formulated using varying combinations of T. occidentalis LPC and soybean meal. These foods were ...

  15. CHEMICAL COMPOSITION AND FUNCTIONAL PROPERTIES OF RICE PROTEIN CONCENTRATES

    Directory of Open Access Journals (Sweden)

    V. V. Kolpakova

    2015-01-01

    Full Text Available Traditionally rice and products of its processing are used to cook porridge, pilaf, lettuce, confectionery, fish, dairy and meat products. At the same time new ways of its processing with releasing of protein products for more effective using, including the use of a glutenfree diet, are developing. The task of this study was a comparative research of nutrition and biological value and functional properties of protein and protein-calcium concentrates produced from rice flour milled from white and brown rice. The traditional and special methods were used. Concentrates were isolated with enzyme preparations of xylanase and amylolytic activity with the next dissolution of protein in diluted hydrochloric acid. Concentrates differed in the content of mineral substances (calcium, zinc, iron and other elements, amino acids and functional properties. The values of the functional properties and indicators of the nutritional value of concentrates from white rice show the advisability of their using in food products, including gluten-free products prepared on the basis of the emulsion and foam systems, and concentrates from brown rice in food products prepared on the basis of using of the emulsion systems. Protein concentrates of brown rice have a low foaming capacity and there is no foam stability at all.

  16. Noninvasive imaging of protein metabolic labeling in single human cells using stable isotopes and Raman microscopy

    NARCIS (Netherlands)

    van Manen, H.J.; Lenferink, Aufrid T.M.; Otto, Cornelis

    2008-01-01

    We have combined nonresonant Raman microspectroscopy and spectral imaging with stable isotope labeling by amino acids in cell culture (SILAC) to selectively detect the incorporation of deuterium-labeled phenylalanine, tyrosine, and methionine into proteins in intact, single HeLa cells. The C−D

  17. Stable Protein-Repellent Zwitterionic Polymer Brushes Grafted from Silicon Nitride

    NARCIS (Netherlands)

    Nguyen, A.T.; Baggerman, J.; Paulusse, J.M.J.; Rijn, van C.J.M.; Zuilhof, H.

    2011-01-01

    Zwitterionic poly(sulfobetaine acrylamide) (SBMAA) brushes were grafted from silicon-rich silicon nitride (SixN4, x > 3) surfaces by atom transfer radical polymerization (ATRP) and studied in protein adsorption experiments. To this aim ATRP initiators were immobilized onto SixN4 through stable

  18. Stable Protein-Repellent Zwitterionic Polymer Brushes Grafted from Silicon Nitride

    NARCIS (Netherlands)

    Nguyen, Ai T.; Baggerman, Jacob; Paulusse, Jos Marie Johannes; van Rijn, Cees J.M.; Zuilhof, Han

    2011-01-01

    Zwitterionic poly(sulfobetaine acrylamide) (SBMAA) brushes were grafted from silicon-rich silicon nitride (SixN4, x > 3) surfaces by atom transfer radical polymerization (ATRP) and studied in protein adsorption experiments. To this aim ATRP initiators were immobilized onto SixN4 through stable Si−C

  19. Continuous protein concentration via free-flow moving reaction boundary electrophoresis.

    Science.gov (United States)

    Kong, Fanzhi; Zhang, Min; Chen, Jingjing; Fan, Liuyin; Xiao, Hua; Liu, Shaorong; Cao, Chengxi

    2017-07-28

    In this work, we developed the model and theory of free-flow moving reaction boundary electrophoresis (FFMRB) for continuous protein concentration for the first time. The theoretical results indicated that (i) the moving reaction boundary (MRB) can be quantitatively designed in free-flow electrophoresis (FFE) system; (ii) charge-to-mass ratio (Z/M) analysis could provide guidance for protein concentration optimization; and (iii) the maximum processing capacity could be predicted. To demonstrate the model and theory, three model proteins of hemoglobin (Hb), cytochrome C (Cyt C) and C-phycocyanin (C-PC) were chosen for the experiments. The experimental results verified that (i) stable MRBs with different velocities could be established in FFE apparatus with weak acid/weak base neutralization reaction system; (ii) proteins of Hb, Cyt C and C-PC were well concentrated with FFMRB; and (iii) a maximum processing capacity and recovery ratio of Cyt C enrichment were 126mL/h and 95.5% respectively, and a maximum enrichment factor was achieved 12.6 times for Hb. All of the experiments demonstrated the protein concentration model and theory. In contrast to other methods, the continuous processing ability enables FFMRB to efficiently enrich diluted protein or peptide in large volume solution. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Use of refractometry for determination of psittacine plasma protein concentration.

    Science.gov (United States)

    Cray, Carolyn; Rodriguez, Marilyn; Arheart, Kristopher L

    2008-12-01

    Previous studies have demonstrated both poor and good correlation of total protein concentrations in various avian species using refractometry and biuret methodologies. The purpose of the current study was to compare these 2 techniques of total protein determination using plasma samples from several psittacine species and to determine the effect of cholesterol and other solutes on refractometry results. Total protein concentration in heparinized plasma samples without visible lipemia was analyzed by refractometry and an automated biuret method on a dry reagent analyzer (Ortho 250). Cholesterol, glucose, and uric acid concentrations were measured using the same analyzer. Results were compared using Deming regression analysis, Bland-Altman bias plots, and Spearman's rank correlation. Correlation coefficients (r) for total protein results by refractometry and biuret methods were 0.49 in African grey parrots (n=28), 0.77 in Amazon parrots (20), 0.57 in cockatiels (20), 0.73 in cockatoos (36), 0.86 in conures (20), and 0.93 in macaws (38) (Prefractometry in Amazon parrots, conures, and macaws (n=25 each, PRefractometry can be used to accurately measure total protein concentration in nonlipemic plasma samples from some psittacine species. Method and species-specific reference intervals should be used in the interpretation of total protein values.

  1. Blood harmane concentrations and dietary protein consumption in essential tremor.

    Science.gov (United States)

    Louis, E D; Zheng, W; Applegate, L; Shi, L; Factor-Litvak, P

    2005-08-09

    Beta-carboline alkaloids (e.g., harmane) are highly tremorogenic chemicals. Animal protein (meat) is the major dietary source of these alkaloids. The authors previously demonstrated that blood harmane concentrations were elevated in patients with essential tremor (ET) vs controls. Whether this difference is due to greater animal protein consumption by patients or their failure to metabolize harmane is unknown. The aim of this study was to determine whether patients with ET and controls differ with regard to 1) daily animal protein consumption and 2) the correlation between animal protein consumption and blood harmane concentration. Data on current diet were collected with a semiquantitative food frequency questionnaire and daily calories and consumption of animal protein and other food types was calculated. Blood harmane concentrations were log-transformed (logHA). The mean logHA was higher in 106 patients than 161 controls (0.61 +/- 0.67 vs 0.43 +/- 0.72 g(-10)/mL, p = 0.035). Patients and controls consumed similar amounts of animal protein (50.2 +/- 19.6 vs 49.4 +/- 19.1 g/day, p = 0.74) and other food types (animal fat, carbohydrates, vegetable fat) and had similar caloric intakes. In controls, logHA was correlated with daily consumption of animal protein (r = 0.24, p = 0.003); in patients, there was no such correlation (r = -0.003, p = 0.98). The similarity between patients and controls in daily animal protein consumption and the absence of the normal correlation between daily animal protein consumption and logHA in patients suggests that another factor (e.g., a metabolic defect) may be increasing blood harmane concentration in patients.

  2. Barrier, mechanical and optical properties of whey protein concentrate films

    Directory of Open Access Journals (Sweden)

    Viviane Machado Azevedo

    2014-08-01

    Full Text Available Whey is recognized as a valuable source of high quality protein and, when processed as protein concentrate, may be used in the production of biodegradable films. The objective of the study was to develop films of whey protein concentrate 80% (WPC at concentrations of 6, 8, 10 and 12% and evaluate the influence of this factor in the barrier, mechanical and optical properties of the films. Treatments showed moisture content with a mean value of 22.10% ± 0.76and high solubility values between 56.67 to 62.42%. Thus, there is little or no influence of varying the concentration of WPC in these properties and high hydrophilicity of the films. With increasing concentration of WPC, increases the water vapor permeability of the films (7.42 x 10-13 to 3.49 x 10-12 g.m-1.s-1.Pa-1. The treatment at the concentration of 6% of WPC showed a higher modulus of elasticity (287.90 ± 41.79 MPa. Thegreater rigidity in films with higher concentrations is possibly due to the greater number of bonds between molecules of the polymeric matrix. The films have the same puncture resistance. The increased concentration of WPC promotes resistance to the action of a localized force. In general, films of whey protein concentrate in the tested concentrations exhibited slightly yellowish color and transparency, and can be used in food packaging that requiring intermediate permeability to water vapor, to keep moisture and texture desired.

  3. Analysis of human protein replacement stable cell lines established using snoMEN-PR vector.

    Directory of Open Access Journals (Sweden)

    Motoharu Ono

    Full Text Available The study of the function of many human proteins is often hampered by technical limitations, such as cytotoxicity and phenotypes that result from overexpression of the protein of interest together with the endogenous version. Here we present the snoMEN (snoRNA Modulator of gene ExpressioN vector technology for generating stable cell lines where expression of the endogenous protein can be reduced and replaced by an exogenous protein, such as a fluorescent protein (FP-tagged version. SnoMEN are snoRNAs engineered to contain complementary sequences that can promote knock-down of targeted RNAs. We have established and characterised two such partial protein replacement human cell lines (snoMEN-PR. Quantitative mass spectrometry was used to analyse the specificity of knock-down and replacement at the protein level and also showed an increased pull-down efficiency of protein complexes containing exogenous, tagged proteins in the protein replacement cell lines, as compared with conventional co-expression strategies. The snoMEN approach facilitates the study of mammalian proteins, particularly those that have so far been difficult to investigate by exogenous expression and has wide applications in basic and applied gene-expression research.

  4. Applications of Protein Thermodynamic Database for Understanding Protein Mutant Stability and Designing Stable Mutants.

    Science.gov (United States)

    Gromiha, M Michael; Anoosha, P; Huang, Liang-Tsung

    2016-01-01

    Protein stability is the free energy difference between unfolded and folded states of a protein, which lies in the range of 5-25 kcal/mol. Experimentally, protein stability is measured with circular dichroism, differential scanning calorimetry, and fluorescence spectroscopy using thermal and denaturant denaturation methods. These experimental data have been accumulated in the form of a database, ProTherm, thermodynamic database for proteins and mutants. It also contains sequence and structure information of a protein, experimental methods and conditions, and literature information. Different features such as search, display, and sorting options and visualization tools have been incorporated in the database. ProTherm is a valuable resource for understanding/predicting the stability of proteins and it can be accessed at http://www.abren.net/protherm/ . ProTherm has been effectively used to examine the relationship among thermodynamics, structure, and function of proteins. We describe the recent progress on the development of methods for understanding/predicting protein stability, such as (1) general trends on mutational effects on stability, (2) relationship between the stability of protein mutants and amino acid properties, (3) applications of protein three-dimensional structures for predicting their stability upon point mutations, (4) prediction of protein stability upon single mutations from amino acid sequence, and (5) prediction methods for addressing double mutants. A list of online resources for predicting has also been provided.

  5. Romanian plant produces protein concentrate from paraffin-nourished yeasts

    Energy Technology Data Exchange (ETDEWEB)

    1985-01-01

    One of the world's few factories in which proteins are produced by continuous biotechnology is located in Romania. Here, at the bioproteins plant, microorganisms are converted into a flour which contains a protein concentrate that is so essential to the fattening of swine, cattle, sheep, fowl, and fish. These microorganisms are Candida type yeasts. The culture medium in which they are grown contains sulfates and phosphates. Paraffin, a petroleum product, supplies the carbon that is essential to the microorganisms viability.

  6. Pea protein concentrate as a substitute for fish meal protein in sea bass diet

    Directory of Open Access Journals (Sweden)

    E. Badini

    2010-01-01

    Full Text Available Pea seeds, even if lower in protein than oilseed meals, have been shown to successfully replace moderate amounts of fish meal protein in diets for carnivorous fish species (Kaushik et al., 1993, Gouveia and Davies, 2000. A further processing of such pulses provides concentrated protein products which look very promising as fish meal substitutes in aquafeeds (Thiessen et al., 2003. The aim of the present study was to evaluate nutrient digestibility, growth response, nutrient and energy retention efficiencies and whole body composition of sea bass (Dicentrarchus labrax, L. fed complete diets in which a pea protein concentrate (PPC was used to replace graded levels of fish meal protein.

  7. The composition and functional properties of whey protein concentrates produced from buttermilk are comparable with those of whey protein concentrates produced from skimmed milk.

    Science.gov (United States)

    Svanborg, Sigrid; Johansen, Anne-Grethe; Abrahamsen, Roger K; Skeie, Siv B

    2015-09-01

    The demand for whey protein is increasing in the food industry. Traditionally, whey protein concentrates (WPC) and isolates are produced from cheese whey. At present, microfiltration (MF) enables the utilization of whey from skim milk (SM) through milk protein fractionation. This study demonstrates that buttermilk (BM) can be a potential source for the production of a WPC with a comparable composition and functional properties to a WPC obtained by MF of SM. Through the production of WPC powder and a casein- and phospholipid (PL)-rich fraction by the MF of BM, sweet BM may be used in a more optimal and economical way. Sweet cream BM from industrial churning was skimmed before MF with 0.2-µm ceramic membranes at 55 to 58°C. The fractionations of BM and SM were performed under the same conditions using the same process, and the whey protein fractions from BM and SM were concentrated by ultrafiltration and diafiltration. The ultrafiltration and diafiltration was performed at 50°C using pasteurized tap water and a membrane with a 20-kDa cut-off to retain as little lactose as possible in the final WPC powders. The ultrafiltrates were subsequently spray dried, and their functional properties and chemical compositions were compared. The amounts of whey protein and PL in the WPC powder from BM (BMWPC) were comparable to the amounts found in the WPC from SM (SMWPC); however, the composition of the PL classes differed. The BMWPC contained less total protein, casein, and lactose compared with SMWPC, as well as higher contents of fat and citric acid. No difference in protein solubility was observed at pH values of 4.6 and 7.0, and the overrun was the same for BMWPC and SMWPC; however, the BMWPC made less stable foam than SMWPC. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  8. Effect of Crude Protein Levels in Concentrate and Concentrate Levels in Diet on Fermentation

    Directory of Open Access Journals (Sweden)

    Dinh Van Dung

    2014-06-01

    Full Text Available The effect of concentrate mixtures with crude protein (CP levels 10%, 13%, 16%, and 19% and diets with roughage to concentrate ratios 80:20, 60:40, 40:60, and 20:80 (w/w were determined on dry matter (DM and organic matter (OM digestibility, and fermentation metabolites using an in vitro fermentation technique. In vitro fermented attributes were measured after 4, 24, and 48 h of incubation respectively. The digestibility of DM and OM, and total volatile fatty acid (VFA increased whereas pH decreased with the increased amount of concentrate in the diet (p<0.001, however CP levels of concentrate did not have any influence on these attributes. Gas production reduced with increased CP levels, while it increased with increasing concentrate levels. Ammonia nitrogen (NH3-N concentration and microbial CP production increased significantly (p<0.05 by increasing CP levels and with increasing concentrate levels in diet as well, however, no significant difference was found between 16% and 19% CP levels. Therefore, 16% CP in concentrate and increasing proportion of concentrate up to 80% in diet all had improved digestibility of DM and organic matter, and higher microbial protein production, with improved fermentation characteristics.

  9. Historical and contemporary stable isotope tracer approaches to studying mammalian protein metabolism

    Science.gov (United States)

    2016-01-01

    Over a century ago, Frederick Soddy provided the first evidence for the existence of isotopes; elements that occupy the same position in the periodic table are essentially chemically identical but differ in mass due to a different number of neutrons within the atomic nucleus. Allied to the discovery of isotopes was the development of some of the first forms of mass spectrometers, driven forward by the Nobel laureates JJ Thomson and FW Aston, enabling the accurate separation, identification, and quantification of the relative abundance of these isotopes. As a result, within a few years, the number of known isotopes both stable and radioactive had greatly increased and there are now over 300 stable or radioisotopes presently known. Unknown at the time, however, was the potential utility of these isotopes within biological disciplines, it was soon discovered that these stable isotopes, particularly those of carbon (13C), nitrogen (15N), oxygen (18O), and hydrogen (2H) could be chemically introduced into organic compounds, such as fatty acids, amino acids, and sugars, and used to “trace” the metabolic fate of these compounds within biological systems. From this important breakthrough, the age of the isotope tracer was born. Over the following 80 yrs, stable isotopes would become a vital tool in not only the biological sciences, but also areas as diverse as forensics, geology, and art. This progress has been almost exclusively driven through the development of new and innovative mass spectrometry equipment from IRMS to GC‐MS to LC‐MS, which has allowed for the accurate quantitation of isotopic abundance within samples of complex matrices. This historical review details the development of stable isotope tracers as metabolic tools, with particular reference to their use in monitoring protein metabolism, highlighting the unique array of tools that are now available for the investigation of protein metabolism in vivo at a whole body down to a single protein level

  10. Whey protein concentration by ultrafiltration and study of functional properties

    Directory of Open Access Journals (Sweden)

    Sidiane Iltchenco

    2018-06-01

    Full Text Available ABSTRACT: This paper aim to evaluate the ultrafiltration (UF process for constituents recovery from whey. Sequences of factorial designs were performed by varying temperature (5 to 40°C and pressure (1 to 3 bar, to maximize the proteins concentration using membrane of 100kDa in dead end system. Based on the best result new experiments were performed with membrane of 50kDa and 10kDa. With the membrane of 50 the protein retention was about 3 times higher than the membrane of 100kDa. The concentrated obtained by UF membrane of 10kDa, 10°C and 2 bar in laboratory scale showed a mean protein retention of 80 %, greater protein solubility, emulsion stability and the identification of β-lactoglobulins (18.3 kDa and α-lactalbumin fractions (14.2kDa. Therefore, the use of membrane of 100 and 50kDa are became a industrially recommendable alternatives to concentration of whey proteins, and/or as a previous step to the fractionation of whey constituents using membrane ≤10kDa, aiming at future applications in different areas (food, pharmaceutical, chemical, etc..

  11. Whey protein concentrate storage at elevated temperature and humidity

    Science.gov (United States)

    Dairy processors are finding new export markets for whey protein concentrate (WPC), a byproduct of cheesemaking, but they need to know if full-sized bags of this powder will withstand high temperature and relative humidity (RH) levels during unrefrigerated storage under tropical conditions. To answ...

  12. Raapzaadeiwitconcentraat en erwteneiwitconcentraat in biologisch biggenvoer = Canola protein concentrate and pea protein concentratrate in diets for organically housed piglets

    NARCIS (Netherlands)

    Peet-Schwering, van der C.M.C.; Binnendijk, G.P.; Diepen, van J.T.M.

    2011-01-01

    At the Experimental Farm Raalte it was investigated whether canola protein concentrate and pea protein concentrate are suitable protein-rich feedstuffs for organically housed piglets. It is concluded that both protein concentrates are suitable protein-rich feedstuffs for piglets. Feed intake and

  13. Using Lead Concentrations and Stable Lead Isotope Ratios to Identify Contamination Events in Alluvial Soils

    Directory of Open Access Journals (Sweden)

    Diane Saint-Laurent

    2010-01-01

    Full Text Available Soils contaminated with hydrocarbons (C10–C50, polycyclic aromatic hydrocarbons (PAHs, and other contaminants (e.g., As, Cd, Cu, Pb were recently discovered on the banks of the Saint-François and Massawippi rivers. Alluvial soils are contaminated over a distance of 100 kilometers, and the level of the contaminated-hydrocarbon layer in the soil profiles is among the highest at the Windsor and Richmond sites. Concentrations of lead and stable lead isotope ratios (204Pb/206Pb, 207Pb/206Pb, 208Pb/206Pb are also used to identify contamination events. The maximum and minimum values detected in soil profiles for arsenic, cadmium, and lead vary from 3.01 to 37.88 mg kg-1 (As, 0.11 to 0.81 mg kg-1 (Cd 12.32 to 149.13 mg kg-1 (Pb, respectively, while the 207Pb/206Pb isotopic ratio values are between 0.8545 and 0.8724 for all the profiles. The highest values of trace elements (As, Pb and Zn were detected in the hydrocarbon layer (C10–C50, most often located at the bottom of the profiles (160, 200, and 220 cm in depth. The various peaks recorded in the soils and the position of the profiles suggest that various contaminants were transported by the river on several occasions and infiltrated the soil matrix or deposited on floodplains during successive floods. Atmospheric particles which entered the river or deposited on riverbanks must also be considered as another source of pollution recorded in soils.

  14. Using Lead Concentrations and Stable Lead Isotope Ratios to Identify Contamination Events in Alluvial Soils

    International Nuclear Information System (INIS)

    Saint-Laurent, D.; St-Laurent, J.; Hahni, M.; Chapados, C.; Ghaleb, B.

    2010-01-01

    Soils contaminated with hydrocarbons (C10,C50), polycyclic aromatic hydrocarbons (PAHs), and other contaminants (e.g., As, Cd, Cu, Pb) were recently discovered on the banks of the Saint-Francois and Massawippi rivers. Alluvial soils are contaminated over a distance of 100 kilometers, and the level of the contaminated-hydrocarbon layer in the soil profiles is among the highest at the Windsor and Richmond sites. Concentrations of lead and stable lead isotope ratios ( 204 Pb/ 206 Pb, 206 Pb/ 207 Pb, 208 Pb/ 20 '6Pb) are also used to identify contamination events. The maximum and minimum values detected in soil profiles for arsenic, cadmium, and lead vary from 3.01 to 37.88?mg kg -1 (As), 0.11 to 0.81?mg kg-1 (Cd) 12.32 to 149.13?mg kg -1 (Pb), respectively, while the 207 Pb/ 206 Pb isotopic ratio values are between 0.8545 and 0.8724 for all the profiles. The highest values of trace elements (As, Pb and Zn) were detected in the hydrocarbon layer (C10,C50), most often located at the bottom of the profiles (160, 200, and 220 cm in depth). The various peaks recorded in the soils and the position of the profiles suggest that various contaminants were transported by the river on several occasions and infiltrated the soil matrix or deposited on flood plains during successive floods. Atmospheric particles which entered the river or deposited on riverbanks must also be considered as another source of pollution recorded in soils

  15. Stable markers of oxidant damage to proteins and their application in the study of human disease

    DEFF Research Database (Denmark)

    Davies, Michael Jonathan; Fu, S; Wang, H

    1999-01-01

    The mechanisms of formation and the nature of the altered amino acid side chains formed on proteins subjected to oxidant attack are reviewed. The use of stable products of protein side chain oxidation as potential markers for assessing oxidative damage in vivo in humans is discussed. The methods...... developed in the authors laboratories are outlined, and the advantages and disadvantages of these techniques compared with other methodologies for assessing oxidative damage to proteins and other macromolecules. Evidence is presented to show that protein oxidation products are sensitive markers of oxidative...... damage, that the pattern of products detected may yield information as to the nature of the original oxidative insult, and that the levels of oxidized side-chains can, in certain circumstances, be much higher than those of other markers of oxidation such as lipid hydroperoxides....

  16. TGP, an extremely stable, non-aggregating fluorescent protein created by structure-guided surface engineering

    Science.gov (United States)

    Close, Devin W.; Don Paul, Craig; Langan, Patricia S.; Wilce, Matthew C.J.; Traore, Daouda A.K.; Halfmann, Randal; Rocha, Reginaldo C.; Waldo, Geoffery S.; Payne, Riley J.; Rucker, Joseph B.; Prescott, Mark; Bradbury, Andrew R.M.

    2014-01-01

    In this paper we describe the engineering and X-ray crystal structure of Thermal Green Protein (TGP), an extremely stable, highly soluble, non-aggregating green fluorescent protein. TGP is a soluble variant of the fluorescent protein eCGP123, which despite being highly stable, has proven to be aggregation-prone. The X-ray crystal structure of eCGP123, also determined within the context of this paper, was used to carry out rational surface engineering to improve its solubility, leading to TGP. The approach involved simultaneously eliminating crystal lattice contacts while increasing the overall negative charge of the protein. Despite intentional disruption of lattice contacts and introduction of high entropy glutamate side chains, TGP crystallized readily in a number of different conditions and the X-ray crystal structure of TGP was determined to 1.9 Å resolution. The structural reasons for the enhanced stability of TGP and eCGP123 are discussed. We demonstrate the utility of using TGP as a fusion partner in various assays and significantly, in amyloid assays in which the standard fluorescent protein, EGFP, is undesirable because of aberrant oligomerization. PMID:25287913

  17. Paracetamol (acetaminophen) protein adduct concentrations during therapeutic dosing.

    Science.gov (United States)

    Heard, Kennon; Green, Jody L; Anderson, Victoria; Bucher-Bartelson, Becki; Dart, Richard C

    2016-03-01

    Paracetamol protein adducts (PPA) are a biomarker of paracetamol exposure. PPA are quantified as paracetamol-cysteine (APAP-CYS), and concentrations above 1.1 μmol l(-1) have been suggested as a marker of paracetamol-induced hepatotoxicity. However, there is little information on the range of concentrations observed during prolonged therapeutic dosing. The aim of the present study was to describe the concentration of PPA in the serum of subjects taking therapeutic doses of paracetamol for at least 16 days. Preplanned secondary aim of a prospective randomized controlled (placebo vs. 4g day(-1) paracetamol) trial. We measured subjects' serum PPA concentrations every 3 days for a minimum of 16 days. We also measured concentrations on study days 1-3 and 16-25 in subsets of patients. PPA were quantified as APAP-CYS after gel filtration and protein digestion using liquid chromatography/mass spectrometry. Ninety per cent of subjects had detectable PPA after five doses. Median APAP-CYS concentrations in paracetamol-treated subjects increased to a plateau of 0.1 μmol l(-1) on day 7, where they remained. The highest concentration measured was 1.1 μmol l(-1) and two subjects never had detectable PPA levels. PPA were detected in the serum of 78% of subjects 9 days after their final dose. PPA are detectable in the vast majority of subjects taking therapeutic doses of paracetamol. While most have concentrations well below the threshold associated with hepatotoxicity, concentrations may approach 1.1 μmol l(-1) in rare cases. Adducts are detectable after a few doses and can persist for over a week after dosing is stopped. © 2015 The British Pharmacological Society.

  18. Highly stable loading of Mcm proteins onto chromatin in living cells requires replication to unload

    Science.gov (United States)

    Kuipers, Marjorie A.; Stasevich, Timothy J.; Sasaki, Takayo; Wilson, Korey A.; Hazelwood, Kristin L.; McNally, James G.; Davidson, Michael W.

    2011-01-01

    The heterohexameric minichromosome maintenance protein complex (Mcm2-7) functions as the eukaryotic helicase during DNA replication. Mcm2-7 loads onto chromatin during early G1 phase but is not converted into an active helicase until much later during S phase. Hence, inactive Mcm complexes are presumed to remain stably bound from early G1 through the completion of S phase. Here, we investigated Mcm protein dynamics in live mammalian cells. We demonstrate that Mcm proteins are irreversibly loaded onto chromatin cumulatively throughout G1 phase, showing no detectable exchange with a gradually diminishing soluble pool. Eviction of Mcm requires replication; during replication arrest, Mcm proteins remained bound indefinitely. Moreover, the density of immobile Mcms is reduced together with chromatin decondensation within sites of active replication, which provides an explanation for the lack of colocalization of Mcm with replication fork proteins. These results provide in vivo evidence for an exceptionally stable lockdown mechanism to retain all loaded Mcm proteins on chromatin throughout prolonged cell cycles. PMID:21220507

  19. Protein concentrate production from the biomass contaminated with radionuclides

    International Nuclear Information System (INIS)

    Nizhko, V.F.; Shinkarenko, M.P.; Polozhaj, V.V.; Krivchik, O.V.

    1992-01-01

    Coefficients of radionuclides accumulation are determined for traditional and rare forage crops grown on contaminated soils. It is shown that with low concentration of radionuclides in soil minimal level of contamination were found in the biomass of lupine (Lupinus luteus L.) and sainfoin (Onobrychis hybridus L.). Relatively high levels of contamination were found in comfrey (Symphytum asperum Lepech.) and bistort (Polygonum divaricatum L.). Comparatively low accumulation coefficients in case of higher density of soil contamination were observed for white and yellow sweetclovers (Melilotus albus Medik. and M. officinalis (L.) Desr.), while higher values of coefficients were found for bird's-foot trefoil (Lotus corniculatus L.), white clover (Trifolium repens L.) and alsike clover (t. hybridum L.). Biomass of white sweet-clover and alsike clover has been processed to produce leaf protein concentrate. It is shown that with biomass contamination of 1 kBq/kg and above conventional technology based on thermal precipitation of the protein does not provide production of pure product. More purified protein concentrates are obtained after two-stage processing of the biomass

  20. Evaluation of behaviour in stabled draught horse foals fed diets with two protein levels.

    Science.gov (United States)

    Sartori, C; Guzzo, N; Normando, S; Bailoni, L; Mantovani, R

    2017-01-01

    The present work is aimed at evaluating the behaviour of Italian Heavy Draught Horse (IHDH) foals reared in semi-covered stables and fed two isoenergetic total mixed rations with different dietary protein levels (13.2% and 10.6% of CP on dry matter). The study was prompted by the restrictions for nitrate emissions in farms of the European Nitrate Directive. One suggested solution is to reduce dietary protein while maintaining normal performance and welfare, but there is a lack of literature in studies of horses. The behaviours of 20 foals of 437±60 kg of BW, aged 379±37 days and stabled in four pens by sex (S) and diet (D) were video recorded and analysed to build a suitable ethogram including 18 behaviours in six categories: ingestion, resting, maintenance, movement, social activities, other. The percentage of the daily time spent in each behavioural category and single behaviours was analysed via a single traits GLM including S, D and their interaction. Daily activity was consistent with existing literature: foals spent about 33% of the day in ingestion activities and 41% in resting, whereas social interactions constituted 8% of the time and individual maintenance draught breeds for foals in both dietary groups, a result that suggests the maintenance of well-being after dietary protein reduction. This result, together with the findings of a companion study showing no changes in growth performances of foals, showed that a reduction of CP in foal diet is reconcilable with the maintenance of performance and welfare.

  1. Protein-lipid interactions in concentrated infant formula

    International Nuclear Information System (INIS)

    Rowley, B.O.; Richardson, T.

    1985-01-01

    Radiolabeled milk proteins ([carbon-14] β-lactoglobulin or [carbon-14] kappa-casein) were added to raw skim milk used to prepare concentrated humanized infant formula. Ultracentrifugation of the sterilized product allowed separation of three fractions: lipids and the proteins associated with them; free casein micelles and other dense particles; and the fluid phase. Distribution of radiolabeled tracer proteins or of protein measured by chemical methods among these three phases varied significantly with differences in processing conditions (time and temperature of sterilization) or amount of certain additives (potassium hydroxide or urea). In the range of 0 to 8 meq/L of potassium hydroxide added to the formula after homogenization but before sterilization, the lipid layer content of carbon-14 from [carbon-14] kappa-casein in the sterilized product decreased by 4.7% for each 1 meq/L of added potassium hydroxide. Lipid layer content of protein decreased by 2 g/L ( of a total of 32 g/L) for each 1 meq/L potassium hydroxide

  2. MALDI-based identification of stable hazelnut protein derived tryptic marker peptides.

    Science.gov (United States)

    Cucu, T; De Meulenaer, B; Devreese, B

    2012-01-01

    Food allergy is an important health problem especially in industrialised countries. Tree nuts, among which are hazelnuts (Corylus avellana), are typically causing serious and life-threatening symptoms in sensitive subjects. Hazelnut is used as a food ingredient in pastry, confectionary products, ice cream and meat products, therefore undeclared hazelnut can be often present as a cross-contaminant representing a threat for allergic consumers. Mass spectrometric techniques are used for the detection of food allergens in processed foods, but limited information regarding stable tryptic peptide markers for hazelnut is available. The aim of this study was to detect stable peptide markers from modified hazelnut protein through the Maillard reaction and oxidation in a buffered solution. Peptides ³⁹⁵Gly-Arg⁴⁰³ from Cor a 11 and ²⁰⁹Gln-Arg²¹⁷, ³⁵¹Ile-Arg³⁶³, ⁴⁶⁴Ala-Arg⁴⁷⁸ and ⁴⁰¹Val-Arg⁴¹⁷ from Cor a 9 hazelnut allergens proved to be the most stable and could be detected and confirmed with high scores in most of the modified samples. The identified peptides can be further used as analytical targets for the development of more robust quantitative methods for hazelnut detection in processed foods.

  3. Assessment of nutritional quality of water hyacinth leaf protein concentrate

    Directory of Open Access Journals (Sweden)

    Oyeyemi Adeyemi

    2016-09-01

    Full Text Available This study was embarked upon to convert water hyacinth, an environmental nuisance, to a natural resource for economic development. Water hyacinth leaf protein concentrate (WHLPC was extracted in edible form and determination of its physicochemical characteristics, total alkaloids and phenolic compounds was done. Analysis of proximate composition and amino acid profile of the WHLPC was also done. The level of heavy metals (mg/kg in WHLPC was found to be Cd (0.02 ± 0.001, Cr (0.13 ± 0.001, Pd (0.003 ± 0.001 and Hg (0.02 ± 0.001 while concentrations of Pb, Pt, Sn, Fe, Cu, Zn, Ni and Co were found to be 0.001 ± 0.00. Level of all heavy metals was found to be within safe limit. Proximate analysis revealed that protein in WHLPC accounted for 50% of its nutrients, carbohydrate accounted for 33% of its nutrients while fat, ash and fibre made up the remaining nutrients. Amino acid analysis showed that WHLPC contained 17 out of 20 common amino acids, particularly, Phe (3.67%, Leu (5.01%. Level of total alkaloids and phenolic compounds was 16.6 mg/kg and 6.0 mg/kg respectively. Evidence from this study suggests that WHLPC is a good source of leaf protein concentrate (LPC; it is nutritious and acutely non toxic.

  4. Tuning calcite morphology and growth acceleration by a rational design of highly stable protein-mimetics

    Science.gov (United States)

    Chen, Chun-Long; Qi, Jiahui; Tao, Jinhui; Zuckermann, Ronald N.; DeYoreo, James J.

    2014-01-01

    In nature, proteins play a significant role in biomineral formation. One of the ultimate goals of bioinspired materials science is to develop highly stable synthetic molecules that mimic the function of these natural proteins by controlling crystal formation. Here, we demonstrate that both the morphology and the degree of acceleration or inhibition observed during growth of calcite in the presence of peptoids can be rationally tuned by balancing the electrostatic and hydrophobic interactions, with hydrophobic interactions playing the dominant role. While either strong electrostatic or hydrophobic interactions inhibit growth and reduces expression of the {104} faces, correlations between peptoid-crystal binding energies and observed changes in calcite growth indicate moderate electrostatic interactions allow peptoids to weakly adsorb while moderate hydrophobic interactions cause disruption of surface-adsorbed water layers, leading to growth acceleration with retained expression of the {104} faces. This study provides fundamental principles for designing peptoids as crystallization promoters, and offers a straightforward screening method based on macroscopic crystal morphology. Because peptoids are sequence-specific, highly stable, and easily synthesized, peptoid-enhanced crystallization offers a broad range of potential applications. PMID:25189418

  5. Distribution of stable free radicals among amino acids of isolated soy proteins.

    Science.gov (United States)

    Lei, Qingxin; Liebold, Christopher M; Boatright, William L; Shah Jahan, M

    2010-09-01

    Application of deuterium sulfide to powdered isolated soy proteins (ISP) was used to quench stable free radicals and produce a single deuterium label on amino acids where free radicals reside. The deuterium labels rendered increases of isotope ratio for the specific ions of radical-bearing amino acids. Isotope ratio measurements were achieved by gas chromatography/mass spectrometry (GC/MS) analyses after the amino acids were released by acidic hydrolysis and converted to volatile derivatives with propyl chloroformate. The isotope enrichment data showed the stable free radicals were located on Ala, Gly, Leu, Ile, Asx (Asp+Asn), Glx (Glu+Gln), and Trp but not on Val, Pro, Met, Phe, Lys, and His. Due to the low abundance of Ser, Thr, and Cys derivatives and the impossibility to accurately measure their isotope ratios, the radical bearing status for these amino acids remained undetermined even though their derivatives were positively identified from ISP hydrolysates. The relative isotope enrichment for radical-bearing amino acids Ala, Gly, Leu, Ile, Asx (Asp+Asn), Glx (Glu+Gln), and Trp were 8.67%, 2.96%, 2.90%, 3.94%, 6.03%, 3.91%, and 21.48%, respectively. Isotope ratio increase for Tyr was also observed but further investigation revealed such increase was mainly from nonspecific deuterium-hydrogen exchange not free radical quenching. The results obtained from the present study provide important information for a better understanding of the mechanisms of free radical formation and stabilization in "dry" ISP.

  6. Stable intermediates determine proteins' primary unfolding sites in the presence of surfactants

    DEFF Research Database (Denmark)

    Petersen, Steen Vang; Andersen, Kell kleiner; Enghild, Jan J.

    2009-01-01

    Despite detailed knowledge of the overall structural changes and stoichiometries of surfactant binding, little is known about which protein regions constitute the preferred sites of attack for initial unfolding. Here we have exposed three proteins to limited proteolysis at anionic (SDS) and catio......Despite detailed knowledge of the overall structural changes and stoichiometries of surfactant binding, little is known about which protein regions constitute the preferred sites of attack for initial unfolding. Here we have exposed three proteins to limited proteolysis at anionic (SDS......) and cationic (DTAC) surfactant concentrations corresponding to specific conformational transitions, using the surfactant-robust broad-specificity proteases Savinase and Alcalase. Cleavage sites are identified by SDS-PAGE and N-terminal sequencing. We observe well-defined cleavage fragments, which suggest......, cleavage sites can be rationalized from the structure of the protein's folding transition state and the position of loops in the native state. Nevertheless, they are more sensitive to choice of surfactant and protease, probably reflecting a heterogeneous and fluctuating ensemble of partially unfolded...

  7. Concentration and stable carbon isotopic composition of CO2 in cave air of Postojnska jama, Slovenia

    Directory of Open Access Journals (Sweden)

    Magda Mandic

    2013-09-01

    Full Text Available Partial pressure of CO2 (pCO2 and its isotopic composition (δ13CairCO2 were measured in Postojnska jama, Slovenia, at 10 locations inside the cave and outside the cave during a one-year period. At all interior locations the pCO2 was higher and δ13CairCO2 lower than in the outside atmosphere. Strong seasonal fluctuations in both parameters were observed at locations deeper in the cave, which are isolated from the cave air circulation. By using a binary mixing model of two sources of CO2, one of them being the atmospheric CO2, we show that the excess of CO2 in the cave air has a δ13C value of -23.3 ± 0.7 ‰, in reasonable agreement with the previously measured soil-CO2 δ13C values. The stable isotope data suggest that soil CO2 is brought to the cave by drip water.

  8. Influence of bleaching on flavor of 34% whey protein concentrate and residual benzoic acid concentration in dried whey products

    Science.gov (United States)

    Previous studies have shown that bleaching negatively affects the flavor of 70% whey protein concentrate (WPC70), but bleaching effects on lower-protein products have not been established. Benzoyl peroxide (BP), a whey bleaching agent, degrades to benzoic acid (BA) and may elevate BA concentrations...

  9. Contribution of glucan-binding protein A to firm and stable biofilm formation by Streptococcus mutans.

    Science.gov (United States)

    Matsumi, Y; Fujita, K; Takashima, Y; Yanagida, K; Morikawa, Y; Matsumoto-Nakano, M

    2015-06-01

    Glucan-binding proteins (Gbps) of Streptococcus mutans, a major pathogen of dental caries, mediate the binding of glucans synthesized from sucrose by the action of glucosyltransferases (GTFs) encoded by gtfB, gtfC, and gtfD. Several stress proteins, including DnaK and GroEL encoded by dnaK and groEL, are related to environmental stress tolerance. The contribution of Gbp expression to biofilm formation was analyzed by focusing on the expression levels of genes encoding GTFs and stress proteins. Biofilm-forming assays were performed using GbpA-, GbpB-, and GbpC-deficient mutant strains and the parental strain MT8148. The expression levels of gtfB, gtfC, gtfD, dnaK, and groEL were evaluated by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Furthermore, the structure of biofilms formed by these Gbp-deficient mutant strains was observed using confocal laser scanning microscopy (CLSM). Biofilm-forming assay findings demonstrated that the amount formed by the GbpA-deficient mutant strain (AD1) was nearly the same as that by the parental strain, while the GbpB- and GbpC-deficient mutant strains produced lower amounts than MT8148. Furthermore, RT-qPCR assay results showed that the expressions of gtfB, dnaK, and groEL in AD1 were elevated compared with MT8148. CLSM also revealed that the structure of biofilm formed by AD1 was prominently different compared with that formed by the parental strain. These results suggest that a defect in GbpA influences the expression of genes controlling biofilm formation, indicating its importance as a protein for firm and stable biofilm formation. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  10. Aggregation in concentrated protein solutions: Insights from rheology, neutron scattering and molecular simulations

    Science.gov (United States)

    Castellanos, Maria Monica

    Aggregation of therapeutic proteins is currently one of the major challenges in the bio-pharmaceutical industry, because aggregates could induce immunogenic responses and compromise the quality of the product. Current scientific efforts, both in industry and academia, are focused on developing rational approaches to screen different drug candidates and predict their stability under different conditions. Moreover, aggregation is promoted in highly concentrated protein solutions, which are typically required for subcutaneous injection. In order to gain further understanding about the mechanisms that lead to aggregation, an approach that combined rheology, neutron scattering, and molecular simulations was undertaken. Two model systems were studied in this work: Bovine Serum Albumin in surfactant-free Phosphate Buffered Saline at pH = 7.4 at concentrations from 11 mg/mL up to ˜519 mg/mL, and a monoclonal antibody in 20 mM Histidine/Histidine Hydrochloride at pH = 6.0 with 60 mg/mL trehalose and 0.2 mg/mL polysorbate-80 at concentrations from 53 mg/mL up to ˜220 mg/mL. The antibody used here has three mutations in the CH2 domain, which result in lower stability upon incubation at 40 °C with respect to the wild-type protein, based on size-exclusion chromatography assays. This temperature is below 49 °C, where unfolding of the least stable, CH2 domain occurs, according to differential scanning calorimetry. This dissertation focuses on identifying the role of aggregation on the viscosity of protein solutions. The protein solutions of this work show an increase in the low shear viscosity in the absence of surfactants, because proteins adsorb at the air/water interface forming a viscoelastic film that affects the measured rheology. Stable surfactant-laden protein solutions behave as simple Newtonian fluids. However, the surfactant-laden antibody solution also shows an increase in the low shear viscosity from bulk aggregation, after prolonged incubation at 40 °C. Small

  11. Protein denaturation and functional properties of Lenient Steam Injection heat treated whey protein concentrate

    DEFF Research Database (Denmark)

    Dickow, Jonatan Ahrens; Kaufmann, Niels; Wiking, Lars

    2012-01-01

    Whey protein concentrate (WPC) was heat treated by use of the novel heat treatment method of Lenient Steam Injection (LSI) to elucidate new functional properties in relation to heat-induced gelation of heat treated WPC. Denaturation was measured by both DSC and FPLC, and the results of the two...... methods were highly correlated. Temperatures of up to 90 °C were applicable using LSI, whereas only 68 °C could be reached by plate heat exchange before coagulation/fouling. Denaturation of whey proteins increased with increasing heat treatment temperature up to a degree of 30–35% denaturation at 90 °C...

  12. Limited hydrolysis of soybean protein concentrate and isolate with ...

    African Journals Online (AJOL)

    STORAGESEVER

    2009-07-20

    Jul 20, 2009 ... world, since its proteins have high biological value while its cost is ... literatures that limited proteolysis of soybean protein pro- ducts offered a ..... hydrolysis of soluble protein present in waste liquors from soy processing.

  13. Using equilibrium passive dosing to maintain stable exposure concentrations of triclosan in a 6-week toxicity test

    DEFF Research Database (Denmark)

    Sobek, A.; Ribbenstedt, A.; Mustajärvi, L.

    2015-01-01

    toxicity tests. Yet, the European Commission’s criteria for chemicals’ risk assessments aim at protecting higher levels in the environment. To achieve protection of populations and ecosystems, reliable long-term ecotoxicologial tests are needed. In this study, we used equilibrium passive dosing to maintain...... stable exposure concentrations of triclosan (log Kow 4.8) in a 6-week multigeneration test with the benthic copepod Nitocra spinipes. The tests were performed in 10 mL vials casted with 1000 mg of silicone (DC 1-2577). Based on a previous pilot study, three triclosan concentrations were selected...... and tested (15 μg L-1; 30 μg L-1; 60 μg L-1) as well as a control (no triclosan). At test beginning, each vial contained 12 individuals consisting of 3 individuals from four different life stages. The test includes feeding with phytoplankton three times a week, which can lead to declining freely dissolved...

  14. Fault tolerance in protein interaction networks: stable bipartite subgraphs and redundant pathways.

    Directory of Open Access Journals (Sweden)

    Arthur Brady

    Full Text Available As increasing amounts of high-throughput data for the yeast interactome become available, more system-wide properties are uncovered. One interesting question concerns the fault tolerance of protein interaction networks: whether there exist alternative pathways that can perform some required function if a gene essential to the main mechanism is defective, absent or suppressed. A signature pattern for redundant pathways is the BPM (between-pathway model motif, introduced by Kelley and Ideker. Past methods proposed to search the yeast interactome for BPM motifs have had several important limitations. First, they have been driven heuristically by local greedy searches, which can lead to the inclusion of extra genes that may not belong in the motif; second, they have been validated solely by functional coherence of the putative pathways using GO enrichment, making it difficult to evaluate putative BPMs in the absence of already known biological annotation. We introduce stable bipartite subgraphs, and show they form a clean and efficient way of generating meaningful BPMs which naturally discard extra genes included by local greedy methods. We show by GO enrichment measures that our BPM set outperforms previous work, covering more known complexes and functional pathways. Perhaps most importantly, since our BPMs are initially generated by examining the genetic-interaction network only, the location of edges in the protein-protein physical interaction network can then be used to statistically validate each candidate BPM, even with sparse GO annotation (or none at all. We uncover some interesting biological examples of previously unknown putative redundant pathways in such areas as vesicle-mediated transport and DNA repair.

  15. Fault tolerance in protein interaction networks: stable bipartite subgraphs and redundant pathways.

    Science.gov (United States)

    Brady, Arthur; Maxwell, Kyle; Daniels, Noah; Cowen, Lenore J

    2009-01-01

    As increasing amounts of high-throughput data for the yeast interactome become available, more system-wide properties are uncovered. One interesting question concerns the fault tolerance of protein interaction networks: whether there exist alternative pathways that can perform some required function if a gene essential to the main mechanism is defective, absent or suppressed. A signature pattern for redundant pathways is the BPM (between-pathway model) motif, introduced by Kelley and Ideker. Past methods proposed to search the yeast interactome for BPM motifs have had several important limitations. First, they have been driven heuristically by local greedy searches, which can lead to the inclusion of extra genes that may not belong in the motif; second, they have been validated solely by functional coherence of the putative pathways using GO enrichment, making it difficult to evaluate putative BPMs in the absence of already known biological annotation. We introduce stable bipartite subgraphs, and show they form a clean and efficient way of generating meaningful BPMs which naturally discard extra genes included by local greedy methods. We show by GO enrichment measures that our BPM set outperforms previous work, covering more known complexes and functional pathways. Perhaps most importantly, since our BPMs are initially generated by examining the genetic-interaction network only, the location of edges in the protein-protein physical interaction network can then be used to statistically validate each candidate BPM, even with sparse GO annotation (or none at all). We uncover some interesting biological examples of previously unknown putative redundant pathways in such areas as vesicle-mediated transport and DNA repair.

  16. Highly concentrated, stable nitrogen-doped graphene for supercapacitors: Simultaneous doping and reduction

    Energy Technology Data Exchange (ETDEWEB)

    Jiang Baojiang [College of Materials Science and Chemical Engineering, Harbin Engineering University, Harbin (China); Key Laboratory of Functional Inorganic Material Chemistry, Ministry of Education of the People' s Republic of China, Heilongjiang University, Harbin (China); Tian Chungui; Wang Lei; Sun Li; Chen Chen; Nong Xiaozhen [Key Laboratory of Functional Inorganic Material Chemistry, Ministry of Education of the People' s Republic of China, Heilongjiang University, Harbin (China); Qiao Yingjie, E-mail: qiaoyingjie@hrbeu.edu.cn [College of Materials Science and Chemical Engineering, Harbin Engineering University, Harbin (China); Fu Honggang, E-mail: fuhg@vip.sina.com [Key Laboratory of Functional Inorganic Material Chemistry, Ministry of Education of the People' s Republic of China, Heilongjiang University, Harbin (China)

    2012-02-01

    In this work, we developed a concentrated ammonia-assisted hydrothermal method to obtain N-doped graphene sheets by simultaneous N-doping and reduction of graphene oxide (GO) sheets. The effects of hydrothermal temperature on the surface chemistry and the structure of N-doped graphene sheets were also investigated. X-ray photoelectron spectroscopy (XPS) study of N-doped graphene reveals that the highest doping level of 7.2% N is achieved at 180 Degree-Sign C for 12 h. N binding configurations of sample consist of pyridine N, quaternary N, and pyridine-N oxides. N doping is accompanied by the reduction of GO with decreases in oxygen levels from 34.8% in GO down to 8.5% in that of N-doped graphene. Meanwhile, the sample exhibits excellent N-doped thermal stability. Electrical measurements demonstrate that products have higher capacitive performance than that of pure graphene, the maximum specific capacitance of 144.6 F/g can be obtained which ascribe the pseudocapacitive effect from the N-doping. The samples also show excellent long-term cycle stability of capacitive performance.

  17. Highly concentrated, stable nitrogen-doped graphene for supercapacitors: Simultaneous doping and reduction

    Science.gov (United States)

    Jiang, Baojiang; Tian, Chungui; Wang, Lei; Sun, Li; Chen, Chen; Nong, Xiaozhen; Qiao, Yingjie; Fu, Honggang

    2012-02-01

    In this work, we developed a concentrated ammonia-assisted hydrothermal method to obtain N-doped graphene sheets by simultaneous N-doping and reduction of graphene oxide (GO) sheets. The effects of hydrothermal temperature on the surface chemistry and the structure of N-doped graphene sheets were also investigated. X-ray photoelectron spectroscopy (XPS) study of N-doped graphene reveals that the highest doping level of 7.2% N is achieved at 180 °C for 12 h. N binding configurations of sample consist of pyridine N, quaternary N, and pyridine-N oxides. N doping is accompanied by the reduction of GO with decreases in oxygen levels from 34.8% in GO down to 8.5% in that of N-doped graphene. Meanwhile, the sample exhibits excellent N-doped thermal stability. Electrical measurements demonstrate that products have higher capacitive performance than that of pure graphene, the maximum specific capacitance of 144.6 F/g can be obtained which ascribe the pseudocapacitive effect from the N-doping. The samples also show excellent long-term cycle stability of capacitive performance.

  18. Atazanavir intracellular concentrations remain stable during pregnancy in HIV-infected patients.

    Science.gov (United States)

    Focà, Emanuele; Calcagno, Andrea; Bonito, Andrea; Simiele, Marco; Domenighini, Elisabetta; D'Avolio, Antonio; Quiros Roldan, Eugenia; Trentini, Laura; Casari, Salvatore; Di Perri, Giovanni; Castelli, Francesco; Bonora, Stefano

    2017-11-01

    Atazanavir (300 mg) boosted by ritonavir (100 mg) is the preferred third drug in pregnancy. However, there is still discordance on atazanavir dose increase during the third trimester. To evaluate plasma and intracellular atazanavir and ritonavir concentrations in HIV-infected women during pregnancy and after delivery. This was an observational study. HIV-infected pregnant patients treated with atazanavir/ritonavir plus either tenofovir/emtricitabine or abacavir/lamivudine had been prospectively enrolled after having signed a written informed consent form. Plasma and intracellular atazanavir and ritonavir Ctrough (24 ± 3 h after drug intake) were measured at each visit during the first, second and third trimesters and post-partum using validated HPLC-MS and HPLC-photodiode array methods (with direct evaluation of cellular volume). Data are described as median (IQR) and compared through non-parametric tests. Twenty-five patients were enrolled; at baseline, the median age was 32 years (27-35). All patients had plasma HIV RNA  0.05). This is the first demonstration that intracellular atazanavir exposure remains unchanged during pregnancy supporting the standard 300/100 mg atazanavir/ritonavir dosing throughout pregnancy. © The Author 2017. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  19. Carbon and nitrogen stable isotope ratios and mercury concentration in the scalp hair of residents from Taiji, a whaling town

    International Nuclear Information System (INIS)

    Endo, Tetsuya; Hayasaka, Moriaki; Hisamichi, Yohsuke; Kimura, Osamu; Haraguchi, Koichi

    2013-01-01

    Highlights: ► We analyzed δ 13 C, δ 15 N and Hg in hair from Japanese whale meat-eaters and non-eaters. ► The δ 15 N and δ 13 C values in whale meat-eaters were higher than those in non-eaters. ► The Hg concentration in whale meat-eaters was higher than that in non-eaters. ► A positive correlation was seen between δ 15 N and Hg in whale meat-eaters. ► Consumption of whale meat may increase δ 15 N, δ 13 C and Hg in the scalp hair. -- Abstract: We analyzed stable isotope ratios of carbon (δ 13 C) and nitrogen (δ 15 N) as well as mercury (Hg) concentration in the scalp hair of Japanese who consumed whale meat and those who did not, and investigated the relationships among the δ 13 C and δ 15 N values and Hg concentration. The average δ 15 N and δ 13 C values of whale meat-eaters (10.11‰ and −18.5‰) were significantly higher than those of non-eaters (9.28‰ and −18.9‰), respectively. The average Hg concentration of whale meat-eaters (20.6 μg/g) was significantly higher than that of non-eaters (2.20 μg/g). Significant positive correlations were found between the δ 13 C and δ 15 N values and between the δ 15 N value and Hg concentration in the hair of whale meat-eaters, while the correlation between the δ 15 N value and Hg concentration was not statistically significant in the non-eaters. The consumption of whale meat may increase Hg concentration as well as δ 15 N and δ 13 C values in scalp hair

  20. Drying and hydration of proteins at high concentration

    NARCIS (Netherlands)

    Bouman, J.

    2015-01-01

    Proteins are the building blocks of life and serve a wide range of essential functions in organisms. Many metabolic reactions in organisms are catalysed by enzymes, DNA is replicated by proteins and in cells proteins often facilitate active transport of e.g. glucose or ions. Proteins also serve

  1. Stable and rigid DTPA-like paramagnetic tags suitable for in vitro and in situ protein NMR analysis.

    Science.gov (United States)

    Chen, Jia-Liang; Zhao, Yu; Gong, Yan-Jun; Pan, Bin-Bin; Wang, Xiao; Su, Xun-Cheng

    2018-02-01

    Organic synthesis of a ligand with high binding affinities for paramagnetic lanthanide ions is an effective way of generating paramagnetic effects on proteins. These paramagnetic effects manifested in high-resolution NMR spectroscopy are valuable dynamic and structural restraints of proteins and protein-ligand complexes. A paramagnetic tag generally contains a metal chelating moiety and a reactive group for protein modification. Herein we report two new DTPA-like tags, 4PS-PyDTTA and 4PS-6M-PyDTTA that can be site-specifically attached to a protein with a stable thioether bond. Both protein-tag adducts form stable lanthanide complexes, of which the binding affinities and paramagnetic tensors are tunable with respect to the 6-methyl group in pyridine. Paramagnetic relaxation enhancement (PRE) effects of Gd(III) complex on protein-tag adducts were evaluated in comparison with pseudocontact shift (PCS), and the results indicated that both 4PS-PyDTTA and 4PS-6M-PyDTTA tags are rigid and present high-quality PREs that are crucially important in elucidation of the dynamics and interactions of proteins and protein-ligand complexes. We also show that these two tags are suitable for in-situ protein NMR analysis.

  2. Study of the proteins in the defatted flour and protein concentrate of baru nuts (Dipteryx alata Vog

    Directory of Open Access Journals (Sweden)

    Rita de Cássia Avellaneda Guimarães

    2012-09-01

    Full Text Available Baru (Dipteryx alata Vog. is an abundant legume in the Brazilian Savanna. Its nuts can be exploited sustainably using its protein and lipid fractions. This study aimed to analyze the proteins of the nuts present in the defatted flour and protein concentrate in terms of their functional properties, the profile of their fractions, and the in vitro digestibility. The flour was defatted with hexane and extracted at the pH of higher protein solubility to obtain the protein concentrate. The electrophoretic profile of the protein fractions was evaluated in SDS-PAGE gel. The functional properties of the proteins indicate the possibility of their use in various foods, like soybeans providing water absorption capacity, oil absorption capacity, emulsifying properties, and foamability. Globulins, followed by the albumins, are the major fractions of the flour and protein concentrate, respectively. Digestibility was greater for the concentrate than for the defatted flour.

  3. Composition and functionality of whey protein phospholipid concentrate and delactosed permeate.

    Science.gov (United States)

    Levin, M A; Burrington, K J; Hartel, R W

    2016-09-01

    Whey protein phospholipid concentrate (WPPC) and delactosed permeate (DLP) are 2 coproducts of cheese whey processing that are currently underused. Past research has shown that WPPC and DLP can be used together as a functional dairy ingredient in foods such as ice cream, soup, and caramel. However, the scope of the research has been limited to 1 WPPC supplier. The objective of this research was to fully characterize a range of WPPC. Four WPPC samples and 1 DLP sample were analyzed for chemical composition and functionality. This analysis showed that WPPC composition was highly variable between suppliers and lots. In addition, the functionality of the WPPC varies depending on the supplier and testing pH, and cannot be correlated with fat or protein content because of differences in processing. The addition of DLP to WPPC affects functionality. In general, WPPC has a high water-holding capacity, is relatively heat stable, has low foamability, and does not aid in emulsion stability. The gel strength and texture are highly dependent on the amount of protein. To be able to use these 2 dairy products, the composition and functionality must be fully understood. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  4. Studies on the functional properties of protein concentrate of Kappaphycus alvarezii (Doty) Doty - an edible seaweed.

    Science.gov (United States)

    Suresh Kumar, K; Ganesan, K; Selvaraj, Kandasamy; Subba Rao, P V

    2014-06-15

    Protein concentrate (PC) of Kappaphycus alvarezii (cultivated on the West coast of India), was extracted and its functional properties were evaluated. The K. alvarezii PC contained 62.3 ± 1.62% proteins. At pH 12, the nitrogen solubility of this PC was 58.72 ± 1.68% in the presence of 0.5M NaCl. The emulsifying and foaming properties of this PC varied with time and pH. However, it formed remarkably stable emulsions with Jatropha oil after 720 min (i.e. E720=53.67 ± 1.59). On the other hand, maximum foaming ability (53.33 ± 2.31%) of the PC was recorded at pH 4.0. This PC had high oil (1.29 ± 0.20 ml oil/g PC) and water absorption capacity (2.22 0.04 ml H2O/g PC). DSC analysis revealed thermal transitions at about 109.25°C at neutral pH. The results obtained in this investigation suggest the suitability of K. alvarezii PC as an inexpensive source of protein; thus this PC could be incorporated into several value-added food products. Copyright © 2013 Elsevier Ltd. All rights reserved.

  5. Distribution and evolution of stable single α-helices (SAH domains in myosin motor proteins.

    Directory of Open Access Journals (Sweden)

    Dominic Simm

    Full Text Available Stable single-alpha helices (SAHs are versatile structural elements in many prokaryotic and eukaryotic proteins acting as semi-flexible linkers and constant force springs. This way SAH-domains function as part of the lever of many different myosins. Canonical myosin levers consist of one or several IQ-motifs to which light chains such as calmodulin bind. SAH-domains provide flexibility in length and stiffness to the myosin levers, and may be particularly suited for myosins working in crowded cellular environments. Although the function of the SAH-domains in human class-6 and class-10 myosins has well been characterised, the distribution of the SAH-domain in all myosin subfamilies and across the eukaryotic tree of life remained elusive. Here, we analysed the largest available myosin sequence dataset consisting of 7919 manually annotated myosin sequences from 938 species representing all major eukaryotic branches using the SAH-prediction algorithm of Waggawagga, a recently developed tool for the identification of SAH-domains. With this approach we identified SAH-domains in more than one third of the supposed 79 myosin subfamilies. Depending on the myosin class, the presence of SAH-domains can range from a few to almost all class members indicating complex patterns of independent and taxon-specific SAH-domain gain and loss.

  6. Identification of syntrophic acetate-oxidizing bacteria in anaerobic digesters by combined protein-based stable isotope probing and metagenomics

    OpenAIRE

    Mosbæk, Freya; Kjeldal, Henrik; Mulat, Daniel G; Albertsen, Mads; Ward, Alastair J; Feilberg, Anders; Nielsen, Jeppe L

    2016-01-01

    Inhibition of anaerobic digestion through accumulation of volatile fatty acids occasionally occurs as the result of unbalanced growth between acidogenic bacteria and methanogens. A fast recovery is a prerequisite for establishing an economical production of biogas. However, very little is known about the microorganisms facilitating this recovery. In this study, we investigated the organisms involved by a novel approach of mapping protein-stable isotope probing (protein-SIP) onto a binned meta...

  7. Nutritional and functional properties of whey proteins concentrate and isolate

    OpenAIRE

    Zoran Herceg; Anet Režek

    2006-01-01

    Whey protein fractions represent 18 - 20 % of total milk nitrogen content. Nutritional value in addition to diverse physico - chemical and functional properties make whey proteins highly suitable for application in foodstuffs. In the most cases, whey proteins are used because of their functional properties. Whey proteins possess favourable functional characteristics such as gelling, water binding, emulsification and foaming ability. Due to application of new process techniques (membrane fract...

  8. Carbon and nitrogen stable isotopes and metal concentration in food webs from a mining-impacted coastal lagoon

    International Nuclear Information System (INIS)

    Marin-Guirao, Lazaro; Lloret, Javier; Marin, Arnaldo

    2008-01-01

    Two food webs from the Mar Menor coastal lagoon, differing in the distance from the desert-stream through which mining wastes were discharged, were examined by reference to essential (Zn and Cu) and non-essential (Pb and Cd) metal concentrations and stable isotopes content (C and N). The partial extraction technique applied, which reflects the availability of metals to organisms after sediment ingestion, showed higher bioavailable metal concentrations in sediments from the station influenced by the mining discharges, in agreement with the higher metal concentrations observed in organisms, which in many cases exceeded the regulatory limits established in Spanish legislation concerning seafood. Spatial differences in essential metal concentrations in the fauna suggest that several organisms are exposed to metal levels above their regulation capacity. Differences in isotopic composition were found between both food webs, the wadi-influenced station showing higher δ 15 N values and lower δ 13 C levels, due to the discharge of urban waste waters and by the entrance of freshwater and allochthonous marsh plants. The linear-regressions between trophic levels (as indicated by δ 15 N) and the metal content indicated that biomagnification does not occur. In the case of invertebrates, since the 'handle strategy' of the species and the physiological requirements of the organisms, among other factors, determine the final concentration of a specific element, no clear relationships between trophic level and the metal content are to be expected. For their part, fish communities did not show clear patterns in the case of any of the analyzed metals, probably because most fish species have similar metal requirements, and because biological factors also intervened. Finally, since the study deals with metals, assumptions concerning trophic transfer factors calculation may not be suitable since the metal burden originates not only from the prey but also from adsorption over the body

  9. Carbon and nitrogen stable isotopes and metal concentration in food webs from a mining-impacted coastal lagoon

    Energy Technology Data Exchange (ETDEWEB)

    Marin-Guirao, Lazaro [Departamento de Ecologia e Hidrologia, Facultad de Biologia, Universidad de Murcia, 30100-Murcia (Spain)], E-mail: lamarin@um.es; Lloret, Javier; Marin, Arnaldo [Departamento de Ecologia e Hidrologia, Facultad de Biologia, Universidad de Murcia, 30100-Murcia (Spain)

    2008-04-01

    Two food webs from the Mar Menor coastal lagoon, differing in the distance from the desert-stream through which mining wastes were discharged, were examined by reference to essential (Zn and Cu) and non-essential (Pb and Cd) metal concentrations and stable isotopes content (C and N). The partial extraction technique applied, which reflects the availability of metals to organisms after sediment ingestion, showed higher bioavailable metal concentrations in sediments from the station influenced by the mining discharges, in agreement with the higher metal concentrations observed in organisms, which in many cases exceeded the regulatory limits established in Spanish legislation concerning seafood. Spatial differences in essential metal concentrations in the fauna suggest that several organisms are exposed to metal levels above their regulation capacity. Differences in isotopic composition were found between both food webs, the wadi-influenced station showing higher {delta}{sup 15}N values and lower {delta}{sup 13}C levels, due to the discharge of urban waste waters and by the entrance of freshwater and allochthonous marsh plants. The linear-regressions between trophic levels (as indicated by {delta}{sup 15}N) and the metal content indicated that biomagnification does not occur. In the case of invertebrates, since the 'handle strategy' of the species and the physiological requirements of the organisms, among other factors, determine the final concentration of a specific element, no clear relationships between trophic level and the metal content are to be expected. For their part, fish communities did not show clear patterns in the case of any of the analyzed metals, probably because most fish species have similar metal requirements, and because biological factors also intervened. Finally, since the study deals with metals, assumptions concerning trophic transfer factors calculation may not be suitable since the metal burden originates not only from the prey but

  10. Delineation of concentration ranges and longitudinal changes of human plasma protein variants.

    Directory of Open Access Journals (Sweden)

    Olgica Trenchevska

    Full Text Available Human protein diversity arises as a result of alternative splicing, single nucleotide polymorphisms (SNPs and posttranslational modifications. Because of these processes, each protein can exists as multiple variants in vivo. Tailored strategies are needed to study these protein variants and understand their role in health and disease. In this work we utilized quantitative mass spectrometric immunoassays to determine the protein variants concentration of beta-2-microglobulin, cystatin C, retinol binding protein, and transthyretin, in a population of 500 healthy individuals. Additionally, we determined the longitudinal concentration changes for the protein variants from four individuals over a 6 month period. Along with the native forms of the four proteins, 13 posttranslationally modified variants and 7 SNP-derived variants were detected and their concentration determined. Correlations of the variants concentration with geographical origin, gender, and age of the individuals were also examined. This work represents an important step toward building a catalog of protein variants concentrations and examining their longitudinal changes.

  11. 24-Hour protein, arginine and citrulline metabolism in fed critically ill children – a stable isotope tracer study

    Science.gov (United States)

    de Betue, Carlijn T.I.; Garcia Casal, Xiomara C.; van Waardenburg, Dick A.; Schexnayder, Stephen M.; Joosten, Koen F.M.; Deutz, Nicolaas E.P.; Engelen, Marielle P.K.J.

    2017-01-01

    Background & aims The reference method to study protein and arginine metabolism in critically ill children is measuring plasma amino acid appearances with stable isotopes during a short (4–8h) time period and extrapolate results to 24-hour. However, 24-hour measurements may be variable due to critical illness related factors and a circadian rhythm could be present. Since only short duration stable isotope studies in critically ill children have been conducted before, the aim of this study was to investigate 24-hour appearance of specific amino acids representing protein and arginine metabolism, with stable isotope techniques in continuously fed critically ill children. Methods In eight critically ill children, admitted to the pediatric (n=4) or cardiovascular (n=4) intensive care unit, aged 0–10 years, receiving continuous (par)enteral nutrition with protein intake 1.0–3.7 g/kg/day, a 24-hour stable isotope tracer protocol was carried out. L-[ring-2H5]-phenylalanine, L-[3,3-2H2]-tyrosine, L-[5,5,5-2H3]-leucine, L-[guanido-15N2]-arginine and L-[5-13C-3,3,4,4-2H4]-citrulline were infused intravenously and L-[15N]-phenylalanine and L-[1-13C]leucine enterally. Arterial blood was sampled every hour. Results Coefficients of variation, representing intra-individual variability, of the amino acid appearances of phenylalanine, tyrosine, leucine, arginine and citrulline were high, on average 14–19% for intravenous tracers and 23–26% for enteral tracers. No evident circadian rhythm was present. The pattern and overall 24-hour level of whole body protein balance differed per individual. Conclusions In continuously fed stable critically ill children, the amino acid appearances of phenylalanine, tyrosine, leucine, arginine and citrulline show high variability. This should be kept in mind when performing stable isotope studies in this population. There was no apparent circadian rhythm. PMID:28089618

  12. Highly Stable Operation of Lithium Metal Batteries Enabled by the Formation of a Transient High Concentration Electrolyte Layer

    Energy Technology Data Exchange (ETDEWEB)

    Zheng, Jianming; Yan, Pengfei; Mei, Donghai; Engelhard, Mark H.; Cartmell, Samuel S.; Polzin, Bryant; Wang, Chong M.; Zhang, Jiguang; Xu, Wu

    2016-02-08

    Lithium (Li) metal has been extensively investigated as an anode for rechargeable battery applications due to its ultrahigh specific capacity and the lowest redox potential. However, significant challenges including dendrite growth and low Coulombic efficiency are still hindering the practical applications of rechargeable Li metal batteries. Here, we demonstrate that long-term cycling of Li metal batteries can be realized by the formation of a transient high concentration electrolyte layer near the surface of Li metal anode during high rate discharge process. The highly concentrated Li+ ions in this transient layer will immediately solvate with the available solvent molecules and facilitate the formation of a stable and flexible SEI layer composed of a poly(ethylene carbonate) framework integrated with other organic/inorganic lithium salts. This SEI layer largely suppresses the corrosion of Li metal anode by free organic solvents and enables the long-term operation of Li metal batteries. The fundamental findings in this work provide a new direction for the development and operation of Li metal batteries that could be operated at high current densities for a wide range of applications.

  13. On the quantitative Amido Black B staining of protein spots in agar gel at low local protein concentrations

    NARCIS (Netherlands)

    Jansen, M.T.

    1962-01-01

    Protein spots in agar gel of identical protein content but different in surface area are found to bind different amounts of dye upon staining with Amido Black B. The lower the protein concentration within the agar gel, the more the Amido Black B content of the spot falls short of the value expected

  14. C-Reactive Protein Concentrations Among Crop and Dairy Farmers with Stable Chronic Obstructive Pulmonary Disease

    Directory of Open Access Journals (Sweden)

    Sasho Stoleski

    2017-09-01

    CONCLUSION: Data obtained suggest that systemic inflammation is present in farmers with COPD and CRP is an important biomarker in COPD in means of reflecting disease severity and prognosis of exposed farmers.

  15. Dry fractionation for production of functional pea protein concentrates

    NARCIS (Netherlands)

    Pelgrom, P.J.M.; Vissers, A.M.; Boom, R.M.; Schutyser, M.A.I.

    2013-01-01

    Dry milling in combination with air classification was evaluated as an alternative to conventional wet extraction of protein from yellow field peas (Pisum sativum). Major advantages of dry fractionation are retention of native functionality of proteins and its lower energy and water use. Peas were

  16. Dry fractionation for sustainable production of plant protein concentrates

    NARCIS (Netherlands)

    Pelgrom, P.J.M.

    2015-01-01

    The global demand for protein-rich foods is expected to double in the coming decades due to the increasing prosperity and world population. To keep up with the demand, the transition from an animal to a plant-based protein supply is desirable from long-term economic and environmental

  17. Dry fractionation for sustainable production of functional legume protein concentrates

    NARCIS (Netherlands)

    Schutyser, M.A.I.; Pelgrom, P.J.M.; Goot, van der A.J.; Boom, R.M.

    2015-01-01

    Plant proteins gain increasing interest as part of a sustainable diet. Because plant materials not only contain protein, they are generally isolated via an energy intensive wet fractionation. This review discusses dry fractionation as an alternative and more sustainable route for producing

  18. Effect of initial protein concentration and pH on in vitro gastric digestion of heated whey proteins.

    Science.gov (United States)

    Zhang, Sha; Vardhanabhuti, Bongkosh

    2014-02-15

    The in vitro digestion of heated whey protein aggregates having different structure and physicochemical properties was evaluated under simulated gastric conditions. Aggregates were formed by heating whey protein isolates (WPI) at 3-9% w/w initial protein concentration and pH 3.0-7.0. Results showed that high protein concentration led to formation of larger WPI aggregates with fewer remaining monomers. Aggregates formed at high protein concentrations showed slower degradation rate compared to those formed at low protein concentration. The effect of initial protein concentration on peptide release pattern was not apparent. Heating pH was a significant factor affecting digestion pattern. At pH above the isoelectric point, the majority of the proteins involved in the aggregation, and aggregates formed at pH 6.0 were more susceptible to pepsin digestion than at pH 7.0. At acidic conditions, only small amount of proteins was involved in the aggregation and heated aggregates were easily digested by pepsin, while the remaining unaggregated proteins were very resistant to gastric digestion. The potential physiological implication of these results on satiety was discussed. Copyright © 2013 Elsevier Ltd. All rights reserved.

  19. New stable isotope method to measure protein digestibility and response to pancreatic enzyme intake in cystic fibrosis.

    Science.gov (United States)

    Engelen, M P K J; Com, G; Anderson, P J; Deutz, N E P

    2014-12-01

    Adequate protein intake and digestion are necessary to prevent muscle wasting in cystic fibrosis (CF). Accurate and easy-to-use methodology to quantify protein maldigestion is lacking in CF. To measure protein digestibility and the response to pancreatic enzyme intake in CF by using a new stable isotope methodology. In 19 CF and 8 healthy subjects, protein digestibility was quantified during continuous (sip) feeding for 6 h by adding (15)N-labeled spirulina protein and L-[ring-(2)H5]phenylalanine (PHE) to the nutrition and measuring plasma ratio [(15)N]PHE to [(2)H5]PHE. Pancreatic enzymes were ingested after 2 h in CF and the response in protein digestibility was assessed. To exclude difference in mucosal function, postabsorptive whole-body citrulline (CIT) production rate was measured by L-[5-(13)C-5,5-(2)H2]-CIT pulse and blood samples were taken to analyze tracer-tracee ratios. Protein digestibility was severely reduced in the CF group (47% of healthy subjects; P digestibility in CF until 90% of values obtained by healthy subjects. Maximal digestibility was reached at 100 min and maintained for 80 min. Stratification into CF children (n = 10) and adults showed comparable values for protein digestibility and similar kinetic responses to pancreatic enzyme intake. Whole-body citrulline production was elevated in CF indicating preserved mucosal function. Protein digestibility is severely compromised in patients with CF as measured by this novel and easy-to-use stable isotope approach. Pancreatic enzymes are able to normalize protein digestibility in CF, albeit with a severe delay. Registration ClinicalTrials.gov = NCT01494909. Copyright © 2013 Elsevier Ltd and European Society for Clinical Nutrition and Metabolism. All rights reserved.

  20. A dual small-molecule rheostat for precise control of protein concentration in Mammalian cells.

    Science.gov (United States)

    Lin, Yu Hsuan; Pratt, Matthew R

    2014-04-14

    One of the most successful strategies for controlling protein concentrations in living cells relies on protein destabilization domains (DD). Under normal conditions, a DD will be rapidly degraded by the proteasome. However, the same DD can be stabilized or "shielded" in a stoichiometric complex with a small molecule, enabling dose-dependent control of its concentration. This process has been exploited by several labs to post-translationally control the expression levels of proteins in vitro as well as in vivo, although the previous technologies resulted in permanent fusion of the protein of interest to the DD, which can affect biological activity and complicate results. We previously reported a complementary strategy, termed traceless shielding (TShld), in which the protein of interest is released in its native form. Here, we describe an optimized protein concentration control system, TTShld, which retains the traceless features of TShld but utilizes two tiers of small molecule control to set protein concentrations in living cells. These experiments provide the first protein concentration control system that results in both a wide range of protein concentrations and proteins free from engineered fusion constructs. The TTShld system has a greatly improved dynamic range compared to our previously reported system, and the traceless feature is attractive for elucidation of the consequences of protein concentration in cell biology. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. The meteorology and chemistry of high nitrogen oxide concentrations in the stable boundary layer at the South Pole

    Science.gov (United States)

    Neff, William; Crawford, Jim; Buhr, Marty; Nicovich, John; Chen, Gao; Davis, Douglas

    2018-03-01

    Four summer seasons of nitrogen oxide (NO) concentrations were obtained at the South Pole (SP) during the Sulfur Chemistry in the Antarctic Troposphere (ISCAT) program (1998 and 2000) and the Antarctic Tropospheric Chemistry Investigation (ANTCI) in (2003, 2005, 2006-2007). Together, analyses of the data collected from these studies provide insight into the large- to small-scale meteorology that sets the stage for extremes in NO and the significant variability that occurs day to day, within seasons, and year to year. In addition, these observations reveal the interplay between physical and chemical processes at work in the stable boundary layer of the high Antarctic plateau. We found a systematic evolution of the large-scale wind system over the ice sheet from winter to summer that controls the surface boundary layer and its effect on NO: initially in early spring (Days 280-310) the transport of warm air and clouds over West Antarctica dominates the environment over the SP; in late spring (Days 310-340), the winds at 300 hPa exhibit a bimodal behavior alternating between northwest and southeast quadrants, which is of significance to NO; in early summer (Days 340-375), the flow aloft is dominated by winds from the Weddell Sea; and finally, during late spring, winds aloft from the southeast are strongly associated with clear skies, shallow stable boundary layers, and light surface winds from the east - it is under these conditions that the highest NO occurs. Examination of the winds at 300 hPa from 1961 to 2013 shows that this seasonal pattern has not changed significantly, although the last twenty years have seen an increasing trend in easterly surface winds at the SP. What has also changed is the persistence of the ozone hole, often into early summer. With lower total ozone column density and higher sun elevation, the highest actinic flux responsible for the photolysis of snow nitrate now occurs in late spring under the shallow boundary layer conditions optimum for

  2. Rapid protein production from stable CHO cell pools using plasmid vector and the cumate gene-switch.

    Science.gov (United States)

    Poulain, Adeline; Perret, Sylvie; Malenfant, Félix; Mullick, Alaka; Massie, Bernard; Durocher, Yves

    2017-08-10

    To rapidly produce large amounts of recombinant proteins, the generation of stable Chinese Hamster Ovary (CHO) cell pools represents a useful alternative to large-scale transient gene expression (TGE). We have developed a cell line (CHO BRI/rcTA ) allowing the inducible expression of recombinant proteins, based on the cumate gene switch. After the identification of optimal plasmid DNA topology (supercoiled vs linearized plasmid) for PEIpro™ mediated transfection and of optimal conditions for methionine sulfoximine (MSX) selection, we were able to generate CHO BRI/rcTA pools producing high levels of recombinant proteins. Volumetric productivities of up to 900mg/L were reproducibly achieved for a Fc fusion protein and up to 350mg/L for an antibody after 14days post-induction in non-optimized fed-batch cultures. In addition, we show that CHO pool volumetric productivities are not affected by a freeze-thaw cycle or following maintenance in culture for over one month in the presence of MSX. Finally, we demonstrate that volumetric protein production with the CR5 cumate-inducible promoter is three- to four-fold higher than with the human CMV or hybrid EF1α-HTLV constitutive promoters. These results suggest that the cumate-inducible CHO BRI/rcTA stable pool platform is a powerful and robust system for the rapid production of gram amounts of recombinant proteins. Crown Copyright © 2017. Published by Elsevier B.V. All rights reserved.

  3. Site-specific tagging proteins with a rigid, small and stable transition metal chelator, 8-hydroxyquinoline, for paramagnetic NMR analysis

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Yin; Huang, Feng [Nankai University, State Key Laboratory of Elemento-Organic Chemistry, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin) (China); Huber, Thomas [Australian National University, Research School of Chemistry (Australia); Su, Xun-Cheng, E-mail: xunchengsu@nankai.edu.cn [Nankai University, State Key Laboratory of Elemento-Organic Chemistry, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin) (China)

    2016-02-15

    Design of a paramagnetic metal binding motif in a protein is a valuable way for understanding the function, dynamics and interactions of a protein by paramagnetic NMR spectroscopy. Several strategies have been proposed to site-specifically tag proteins with paramagnetic lanthanide ions. Here we report a simple approach of engineering a transition metal binding motif via site-specific labelling of a protein with 2-vinyl-8-hydroxyquinoline (2V-8HQ). The protein-2V-8HQ adduct forms a stable complex with transition metal ions, Mn(II), Co(II), Ni(II), Cu(II) and Zn(II). The paramagnetic effects generated by these transition metal ions were evaluated by NMR spectroscopy. We show that 2V-8HQ is a rigid and stable transition metal binding tag. The coordination of the metal ion can be assisted by protein sidechains. More importantly, tunable paramagnetic tensors are simply obtained in an α-helix that possesses solvent exposed residues in positions i and i + 3, where i is the residue to be mutated to cysteine, i + 3 is Gln or Glu or i − 4 is His. The coordination of a sidechain carboxylate/amide or imidazole to cobalt(II) results in different structural geometries, leading to different paramagnetic tensors as shown by experimental data.

  4. Interactive Effects of CO2 Concentration and Water Regime on Stable Isotope Signatures, Nitrogen Assimilation and Growth in Sweet Pepper

    Directory of Open Access Journals (Sweden)

    María D. Serret

    2018-01-01

    Full Text Available Sweet pepper is among the most widely cultivated horticultural crops in the Mediterranean basin, being frequently grown hydroponically under cover in combination with CO2 fertilization and water conditions ranging from optimal to suboptimal. The aim of this study is to develop a simple model, based on the analysis of plant stable isotopes in their natural abundance, gas exchange traits and N concentration, to assess sweet pepper growth. Plants were grown in a growth chamber for near 6 weeks. Two [CO2] (400 and 800 μmol mol−1, three water regimes (control and mild and moderate water stress and four genotypes were assayed. For each combination of genotype, [CO2] and water regime five plants were evaluated. Water stress applied caused significant decreases in water potential, net assimilation, stomatal conductance, intercellular to atmospheric [CO2], and significant increases in water use efficiency, leaf chlorophyll content and carbon isotope composition, while the relative water content, the osmotic potential and the content of anthocyanins did change not under stress compared to control conditions support this statement. Nevertheless, water regime affects plant growth via nitrogen assimilation, which is associated with the transpiration stream, particularly at high [CO2], while the lower N concentration caused by rising [CO2] is not associated with stomatal closure. The stable isotope composition of carbon, oxygen, and nitrogen (δ13C, δ18O, and δ15N in plant matter are affected not only by water regime but also by rising [CO2]. Thus, δ18O increased probably as response to decreases in transpiration, while the increase in δ15N may reflect not only a lower stomatal conductance but a higher nitrogen demand in leaves or shifts in nitrogen metabolism associated with decreases in photorespiration. The way that δ13C explains differences in plant growth across water regimes within a given [CO2], seems to be mediated through its direct

  5. Spray dried microparticles of chia oil using emulsion stabilized by whey protein concentrate and pectin by electrostatic deposition.

    Science.gov (United States)

    Noello, C; Carvalho, A G S; Silva, V M; Hubinger, M D

    2016-11-01

    Chia seed oil has a high content of α-linolenic acid (60%) and linoleic acid (20%). Use of this oil in different products is limited due to its liquid state, and the presence of insaturation is a trigger for oxidation. In this context, to facilitate the incorporation of chia oil in food products and increase its protection against oxidation, the aim of this work was to produce chia oil microparticles by spray drying using emulsions stabilized by whey protein concentrate (ζ-potential +13.4 at pH3.8) and pectin (ζ-potential -40.4 at pH3.8) through the electrostatic layer-by-layer deposition technique and emulsions prepared with only whey protein concentrate. Emulsions stabilized by whey protein concentrate and stabilized by whey protein concentrate-pectin were prepared using maltodextrin (10 DE) and modified starch (Hi-Cap® 100). They were characterized in relation to stability, droplet size, ζ-Potential and optical microscopy. The microparticles were characterized in relation to moisture content, water activity, particle size, microstructure and oxidative stability by the Rancimat method. Emulsions stabilized by whey protein concentrate-pectin with added maltodextrin 10 DE and emulsions stabilized by whey protein concentrate with added modified starch (Hi-Cap® 100) were stable after 24h. Emulsions stabilized by whey protein concentrate and by whey protein concentrate-pectin showed droplets with mean diameter ranging from 0.80 to 1.31μm, respectively and ζ-potential varying from -6.9 to -27.43mV, respectively. After spray drying, the microparticles showed an mean diameter ranging from 7.00 to 9.00μm. All samples presented high encapsulation efficiency values, above 99%. Microparticles produced with modified starch showed a smoother spherical surface than particles with maltodextrin 10 DE, which presented a wrinkled surface. All microparticles exhibited higher oxidative stability than chia oil in pure form. Copyright © 2016 Elsevier Ltd. All rights reserved.

  6. Radiation-induced dissociation of stable DNA-protein complexes in Erlich ascites carcinoma cells

    International Nuclear Information System (INIS)

    Juhasz, P.P.; Sirota, N.P.; Gaziev, A.I.

    1982-01-01

    DNA of Ehrlich ascites carcinoma cells prepared under conditions that were highly denaturing for proteins but not for DNA, contained a group of nonhistone residual proteins. The amount of these proteins increased during DNA replication. The DNA-protein complex observed was sensitive to proteolytic enzymes and/or SH-reagents. γ-irradiation cells with moderate doses leads to a decrease in the amount of DNA-protein complexes. High-dose gamma-irradiation produces enhanced linking of chromosomal proteins with DNA. (author)

  7. High-Temperature Thermochemical Storage with Redox-Stable Perovskites for Concentrating Solar Power, CRADA Number: CRD-14-554

    Energy Technology Data Exchange (ETDEWEB)

    Ma, Zhiwen [National Renewable Energy Laboratory (NREL), Golden, CO (United States)

    2017-09-05

    As part of a Federal Opportunity Announcement (FOA) Award, the project will be led by Colorado School of Mines (CSM) to explore and demonstrate the efficacy of highly reducible, redox-stable oxides to provide efficient thermochemical energy storage for heat release at temperatures of 900 degrees Celcius or more. NREL will support the material development for its application in a concentrating solar power (CSP) plant. In the project, NREL will provide its inventive system design, chemical looping for CSP, and use it as a platform to accommodate the chemical processes using a cost effective perovskite materials identified by CSM. NREL will design a 5-10kW particle receiver for perovskite reduction to store solar energy and help the development of a fluidized-bed reoxidation reactor and system integration. NREL will develop the demonstration receiver for on-sun test in the 5-10 kWt range in NREL's high flux solar furnace. NREL will assist in system analysis and provide techno-economic inputs for the overall system configuration.

  8. Amino Acid Composition of an Organic Brown Rice Protein Concentrate and Isolate Compared to Soy and Whey Concentrates and Isolates

    Directory of Open Access Journals (Sweden)

    Douglas S. Kalman

    2014-06-01

    Full Text Available A protein concentrate (Oryzatein-80™ and a protein isolate (Oryzatein-90™ from organic whole-grain brown rice were analyzed for their amino acid composition. Two samples from different batches of Oryzatein-90™ and one sample of Oryzatein-80™ were provided by Axiom Foods (Los Angeles, CA, USA. Preparation and analysis was carried out by Covance Laboratories (Madison, WI, USA. After hydrolysis in 6-N hydrochloric acid for 24 h at approximately 110 °C and further chemical stabilization, samples were analyzed by HPLC after pre-injection derivitization. Total amino acid content of both the isolate and the concentrate was approximately 78% by weight with 36% essential amino acids and 18% branched-chain amino acids. These results are similar to the profiles of raw and cooked brown rice except in the case of glutamic acid which was 3% lower in the isolate and concentrate. The amino acid content and profile of the Oryzatein-90™ isolate was similar to published values for soy protein isolate but the total, essential, and branched-chain amino acid content of whey protein isolate was 20%, 39% and 33% greater, respectively, than that of Oryzatein-90™. These results provide a valuable addition to the nutrient database of protein isolates and concentrates from cereal grains.

  9. Amino Acid Composition of an Organic Brown Rice Protein Concentrate and Isolate Compared to Soy and Whey Concentrates and Isolates.

    Science.gov (United States)

    Kalman, Douglas S

    2014-06-30

    A protein concentrate (Oryzatein-80™) and a protein isolate (Oryzatein-90™) from organic whole-grain brown rice were analyzed for their amino acid composition. Two samples from different batches of Oryzatein-90™ and one sample of Oryzatein-80™ were provided by Axiom Foods (Los Angeles, CA, USA). Preparation and analysis was carried out by Covance Laboratories (Madison, WI, USA). After hydrolysis in 6-N hydrochloric acid for 24 h at approximately 110 °C and further chemical stabilization, samples were analyzed by HPLC after pre-injection derivitization. Total amino acid content of both the isolate and the concentrate was approximately 78% by weight with 36% essential amino acids and 18% branched-chain amino acids. These results are similar to the profiles of raw and cooked brown rice except in the case of glutamic acid which was 3% lower in the isolate and concentrate. The amino acid content and profile of the Oryzatein-90™ isolate was similar to published values for soy protein isolate but the total, essential, and branched-chain amino acid content of whey protein isolate was 20%, 39% and 33% greater, respectively, than that of Oryzatein-90™. These results provide a valuable addition to the nutrient database of protein isolates and concentrates from cereal grains.

  10. Deciphering systemic wound responses of the pumpkin extrafascicular phloem by metabolomics and stable isotope-coded protein labeling.

    Science.gov (United States)

    Gaupels, Frank; Sarioglu, Hakan; Beckmann, Manfred; Hause, Bettina; Spannagl, Manuel; Draper, John; Lindermayr, Christian; Durner, Jörg

    2012-12-01

    In cucurbits, phloem latex exudes from cut sieve tubes of the extrafascicular phloem (EFP), serving in defense against herbivores. We analyzed inducible defense mechanisms in the EFP of pumpkin (Cucurbita maxima) after leaf damage. As an early systemic response, wounding elicited transient accumulation of jasmonates and a decrease in exudation probably due to partial sieve tube occlusion by callose. The energy status of the EFP was enhanced as indicated by increased levels of ATP, phosphate, and intermediates of the citric acid cycle. Gas chromatography coupled to mass spectrometry also revealed that sucrose transport, gluconeogenesis/glycolysis, and amino acid metabolism were up-regulated after wounding. Combining ProteoMiner technology for the enrichment of low-abundance proteins with stable isotope-coded protein labeling, we identified 51 wound-regulated phloem proteins. Two Sucrose-Nonfermenting1-related protein kinases and a 32-kD 14-3-3 protein are candidate central regulators of stress metabolism in the EFP. Other proteins, such as the Silverleaf Whitefly-Induced Protein1, Mitogen Activated Protein Kinase6, and Heat Shock Protein81, have known defensive functions. Isotope-coded protein labeling and western-blot analyses indicated that Cyclophilin18 is a reliable marker for stress responses of the EFP. As a hint toward the induction of redox signaling, we have observed delayed oxidation-triggered polymerization of the major Phloem Protein1 (PP1) and PP2, which correlated with a decline in carbonylation of PP2. In sum, wounding triggered transient sieve tube occlusion, enhanced energy metabolism, and accumulation of defense-related proteins in the pumpkin EFP. The systemic wound response was mediated by jasmonate and redox signaling.

  11. Usefulness of C1 Esterase Inhibitor Protein Concentrate in the ...

    African Journals Online (AJOL)

    2018-04-04

    Apr 4, 2018 ... 2018 Nigerian Journal of Clinical Practice | Published by Wolters Kluwer ‑ Medknow ... of this case report is to describe the lifesaving use of a novel C1‑INH protein ... edema of the upper lip, uvula, and tongue [Figure 1].

  12. Intestinal DNA concentration and protein synthesis in response to ...

    African Journals Online (AJOL)

    Performance, protein synthesis and mucosal DNA in small intestine of Leghorn hens may be affected by low quality feedstuff. An experiment was conducted in completely randomized design (CRD) in 2 × 2 factorial arrangement. Main factors included diets containing 20 and 40 % barley and black and blue strains of leghorn ...

  13. Intestinal DNA concentration and protein synthesis in response to ...

    African Journals Online (AJOL)

    Jane

    2011-10-05

    Oct 5, 2011 ... Full Length Research Paper. Intestinal ... transporters are membrane-bound proteins and operate ... sporters that are similar to those found on other plasma ... on fastDNA® kit (application manual revision 6540-400-4H01) and.

  14. Age-dependent changes in the total protein concentrations in the ...

    African Journals Online (AJOL)

    related changes in total protein concentrations in ten regions of the pig brain and hypophyses from birth to 36 months of age. Age-related changes in protein concentrations in all the brain regions except the pons and cerebral cortex were not ...

  15. DNA-, RNA-, and Protein-Based Stable-Isotope Probing for High-Throughput Biomarker Analysis of Active Microorganisms.

    Science.gov (United States)

    Jameson, Eleanor; Taubert, Martin; Coyotzi, Sara; Chen, Yin; Eyice, Özge; Schäfer, Hendrik; Murrell, J Colin; Neufeld, Josh D; Dumont, Marc G

    2017-01-01

    Stable-isotope probing (SIP) enables researchers to target active populations within complex microbial communities, which is achieved by providing growth substrates enriched in heavy isotopes, usually in the form of 13 C, 18 O, or 15 N. After growth on the substrate and subsequent extraction of microbial biomarkers, typically nucleic acids or proteins, the SIP technique is used for the recovery and analysis of isotope-labeled biomarkers from active microbial populations. In the years following the initial development of DNA- and RNA-based SIP, it was common practice to characterize labeled populations by targeted gene analysis. Such approaches usually involved fingerprint-based analyses or sequencing of clone libraries containing 16S rRNA genes or functional marker gene amplicons. Although molecular fingerprinting remains a valuable approach for rapid confirmation of isotope labeling, recent advances in sequencing technology mean that it is possible to obtain affordable and comprehensive amplicon profiles, metagenomes, or metatranscriptomes from SIP experiments. Not only can the abundance of microbial groups be inferred from metagenomes, but researchers can bin, assemble, and explore individual genomes to build hypotheses about the metabolic capabilities of labeled microorganisms. Analysis of labeled mRNA is a more recent advance that can provide independent metatranscriptome-based analysis of active microorganisms. The power of metatranscriptomics is that mRNA abundance often correlates closely with the corresponding activity of encoded enzymes, thus providing insight into microbial metabolism at the time of sampling. Together, these advances have improved the sensitivity of SIP methods and allow the use of labeled substrates at ecologically relevant concentrations. Particularly as methods improve and costs continue to drop, we expect that the integration of SIP with multiple omics-based methods will become prevalent components of microbial ecology studies

  16. Physical and chemical changes in whey protein concentrate stored at elevated temperature and humidity

    Science.gov (United States)

    The chemistry of whey protein concentrate (WPC) under adverse storage conditions was monitored to provide information on shelf life in hot, humid areas. WPC34 (34.9 g protein/100 g) and WPC80 (76.8 g protein/100 g) were stored for up to 18 mo under ambient conditions and at elevated temperature and...

  17. 21 CFR 184.1979c - Whey protein concentrate.

    Science.gov (United States)

    2010-04-01

    ... the methods prescribed in section 16.057 (liquid sample), entitled “Gravimetric Method—Official Final... concentrate, on a dry product basis, based on analytical methods in the referenced sections of “Official Methods of Analysis of the Association of Official Analytical Chemists,” 13th ed. (1980), which is...

  18. Thermal-stable proteins of fruit of long-living Sacred Lotus Nelumbo nucifera Gaertn var. China Antique.

    Science.gov (United States)

    Shen-Miller, J; Lindner, Petra; Xie, Yongming; Villa, Sarah; Wooding, Kerry; Clarke, Steven G; Loo, Rachel R O; Loo, Joseph A

    2013-09-01

    Single-seeded fruit of the sacred lotus Nelumbo nucifera Gaertn var. China Antique from NE China have viability as long as ~1300 years determined by direct radiocarbon-dating, having a germination rate of 84%. The pericarp, a fruit tissue that encloses the single seeds of Nelumbo , is considered one of the major factors that contribute to fruit longevity. Proteins that are heat stable and have protective function may be equally important to seed viability. We show proteins of Nelumbo fruit that are able to withstand heating, 31% of which remained soluble in the 110°C-treated embryo-axis of a 549-yr-old fruit and 76% retained fluidity in its cotyledons. Genome of Nelumbo is published. The amino-acid sequences of 11 "thermal proteins" (soluble at 100°C) of modern Nelumbo embryo-axes and cotyledons, identified by mass spectrometry, Western blot and bioassay, are assembled and aligned with those of an archaeal-hyperthermophile Methancaldococcus jannaschii (Mj; an anaerobic methanogen having a growth optimum of 85°C) and with five mesophile angiosperms. These thermal proteins have roles in protection and repair under stress. More than half of the Nelumbo thermal proteins (55%) are present in the archaean Mj, indicating their long-term durability and history. One Nelumbo protein-repair enzyme exhibits activity at 100°C, having a higher heat-tolerance than that of Arabidopsis. A list of 30 sequenced but unassembled thermal proteins of Nelumbo is supplemented.

  19. Evaluation of energy status of dairy cows using milk fat, protein and urea concentrations

    Directory of Open Access Journals (Sweden)

    Kirovski Danijela

    2011-11-01

    Full Text Available Energy status of dairy cows may be estimated using results for concentrations of fat, protein and urea (MUN in milk samples obtained from bulk tank or individual cows. Using individual cow milk samples is recommended on dairy farms in our geografical region due to the unhomogenity of cows in the herds in respect to their genetic potential for milk production. Depression of milk fat occurs as a consequence of heat stress, underfeeding of peripartal cows, overfeeding concentrate with reduced ration fiber levels or overfeeding with dietary fat. High milk fat content is usually combined with severe negative energy balance. Nutrition and feeding practices have great impact on milk protein level. A deficiency of crude protein in the ration may depress protein in milk. Feeding excessive dietary protein does not significantly increase milk protein. MUN analyses point out potential problems in feeding program on dairy farm. High MUN values may reflect excessive dietary crude protein and/or low rumen degradable non fiber carbohydrates intake. Also, MUN levels is impacted by heat stress since its value is increased during the summer season. Low MUNs indicate a possible dietary protein deficiency. Additionally, low MUNs concentration may indicate excess in dietary nonstructural carbohydrates. On the bases on the interrelationships between protein and urea concentrations, as well as protein and fat concentrations in individual milk sample, estimation of energy balance of dairy cows may be done more accurately.

  20. Biomimetic triblock copolymer membrane arrays: a stable template for functional membrane proteins

    DEFF Research Database (Denmark)

    Gonzalez-Perez, A.; Jensen, Karin Bagger Stibius; Vissing, Thomas

    2009-01-01

    It is demonstrated that biomimetic stable triblock copolymer membrane arrays can be prepared using a scaffold containing 64 apertures of 300 μm diameter each. The membranes were made from a stock solution of block copolymers with decane as a solvent using a new deposition method. By using decane...

  1. The promyelocytic leukemia gene product (PML) forms stable complexes with the retinoblastoma protein

    DEFF Research Database (Denmark)

    Alcalay, M; Tomassoni, L; Colombo, E

    1998-01-01

    PML is a nuclear protein with growth-suppressive properties originally identified in the context of the PML-retinoic acid receptor alpha (RAR alpha) fusion protein of acute promyelocytic leukemia. PML localizes within distinct nuclear structures, called nuclear bodies, which are disrupted by the ...

  2. A robust and rapid method of producing soluble, stable, and functional G-protein coupled receptors.

    Directory of Open Access Journals (Sweden)

    Karolina Corin

    Full Text Available Membrane proteins, particularly G-protein coupled receptors (GPCRs, are notoriously difficult to express. Using commercial E. coli cell-free systems with the detergent Brij-35, we could rapidly produce milligram quantities of 13 unique GPCRs. Immunoaffinity purification yielded receptors at >90% purity. Secondary structure analysis using circular dichroism indicated that the purified receptors were properly folded. Microscale thermophoresis, a novel label-free and surface-free detection technique that uses thermal gradients, showed that these receptors bound their ligands. The secondary structure and ligand-binding results from cell-free produced proteins were comparable to those expressed and purified from HEK293 cells. Our study demonstrates that cell-free protein production using commercially available kits and optimal detergents is a robust technology that can be used to produce sufficient GPCRs for biochemical, structural, and functional analyses. This robust and simple method may further stimulate others to study the structure and function of membrane proteins.

  3. Biochemical studies on gamma irradiated male rats fed on whey protein concentrate

    International Nuclear Information System (INIS)

    Mohamed, N.E; Anwar, M.M.; El-bostany, N.A.

    2010-01-01

    This study carried out to investigate the possible role of whey protein protein concentrate in ameliorating some biochemical disorders induced in gamma irradiated male rats. Forty eight male albino rats were divided into four equal groups: Group 1 fed on normal diet during experimental period. Group 2 where the diet contain 15 % whey protein concentrate instead of soybean protein . Group 3 rats were exposed to whole body gamma radiation with single dose of 5 Gy and fed on the normal diet. Group 4 rate exposed to 5 Gy then fed on diet contain 15 % whey protein concentrate, the rats were decapitated after two and four weeks post irradiation. Exposure to whole body irradiation caused significant elevation of serum ALT, AST, glucose, urea, creatinine and total triiodothyronine with significant decrease in total protein, albumin and thyroxin. Irradiated rats fed on whey protein concentrate revealed significant improvement of some biochemical parameters. It could be conclude that whey protein concentrate may be considered as a useful protein source for reducing radiation injury via metabolic pathway.

  4. Composition, structure and functional properties of protein concentrates and isolates produced from walnut (Juglans regia L.).

    Science.gov (United States)

    Mao, Xiaoying; Hua, Yufei

    2012-01-01

    In this study, composition, structure and the functional properties of protein concentrate (WPC) and protein isolate (WPI) produced from defatted walnut flour (DFWF) were investigated. The results showed that the composition and structure of walnut protein concentrate (WPC) and walnut protein isolate (WPI) were significantly different. The molecular weight distribution of WPI was uniform and the protein composition of DFWF and WPC was complex with the protein aggregation. H(0) of WPC was significantly higher (p structure of WPI was similar to WPC. WPI showed big flaky plate like structures; whereas WPC appeared as a small flaky and more compact structure. The most functional properties of WPI were better than WPC. In comparing most functional properties of WPI and WPC with soybean protein concentrate and isolate, WPI and WPC showed higher fat absorption capacity (FAC). Emulsifying properties and foam properties of WPC and WPI in alkaline pH were comparable with that of soybean protein concentrate and isolate. Walnut protein concentrates and isolates can be considered as potential functional food ingredients.

  5. Effect of Addition of Concentrated Proteins and Seminal Plasma Low Molecular Weight Proteins in Freezing and Thawing of Equine Semen

    Directory of Open Access Journals (Sweden)

    Fagundes, B.

    2011-07-01

    Full Text Available Difficulties in obtaining equine frozen semen with potential fertility are recognized. This study was designed to investigate the effect of seminal plasma on frozen/thawing of eight stallion semen from different breed using the following treatments: Seminal plasma with ten-fold concentrated proteins with molecular weight above 10 kDa on frozen extender; Part of seminal plasma with proteins under 10 kDa on frozen extender; Conventional freezing, using whole seminal plasma on frozen extender. Using the parameter of 30% of seminal motility post-thawing as index of good freezability, it was verified an increased percentage of stallions that presented good freezability when semen was frozen with seminal plasma containing ten-fold concentrated proteins with molecular weight above 10 kDa on frozen extender. These results, suggested the use of seminal plasma concentrated proteins from own stallion to freezing/thawing semen.

  6. Concentration of serum thyroid hormone binding proteins after 131I treatment of hyperthyroidism

    International Nuclear Information System (INIS)

    Harrop, J.S.; Hopton, M.R.; Lazarus, J.H.

    1981-01-01

    Serum concentrations of the thyroid hormone binding proteins, thyroxine binding globulin, prealbumin, and albumin were determined in 30 thyrotoxic patients before and after 131 I treatment. Each patient was placed into one of three groups according to response to treatment. The serum concentration of all three proteins rose significantly in 10 patients who became euthyroid, and a greater increase was seen in 10 patients who developed hypothyroidism. There was no significant change in thyroid hormone binding protein concentrations in 10 subjects who remained hyperthyroid. Changes in the concentration of thyroid hormone binding proteins should be borne in mind when total thyroid hormone concentrations are used to monitor the progress of patients receiving treatment for hyperthyroidism. (author)

  7. Expression of a Recombinant Anti-HIV and Anti-Tumor Protein, MAP30, in Nicotiana tobacum Hairy Roots: A pH-Stable and Thermophilic Antimicrobial Protein.

    Directory of Open Access Journals (Sweden)

    Ali Moghadam

    Full Text Available In contrast to conventional antibiotics, which microorganisms can readily evade, it is nearly impossible for a microbial strain that is sensitive to antimicrobial proteins to convert to a resistant strain. Therefore, antimicrobial proteins and peptides that are promising alternative candidates for the control of bacterial infections are under investigation. The MAP30 protein of Momordica charantia is a valuable type I ribosome-inactivating protein (RIP with anti-HIV and anti-tumor activities. Whereas the antimicrobial activity of some type I RIPs has been confirmed, less attention has been paid to the antimicrobial activity of MAP30 produced in a stable, easily handled, and extremely cost-effective protein-expression system. rMAP30-KDEL was expressed in Nicotiana tobacum hairy roots, and its effect on different microorganisms was investigated. Analysis of the extracted total proteins of transgenic hairy roots showed that rMAP30-KDEL was expressed effectively and that this protein exhibited significant antibacterial activity in a dose-dependent manner. rMAP30-KDEL also possessed thermal and pH stability. Bioinformatic analysis of MAP30 and other RIPs regarding their conserved motifs, amino-acid contents, charge, aliphatic index, GRAVY value, and secondary structures demonstrated that these factors accounted for their thermophilicity. Therefore, RIPs such as MAP30 and its derived peptides might have promising applications as food preservatives, and their analysis might provide useful insights into designing clinically applicable antibiotic agents.

  8. Cell-free expression and stable isotope labelling strategies for membrane proteins

    International Nuclear Information System (INIS)

    Sobhanifar, Solmaz; Reckel, Sina; Junge, Friederike; Schwarz, Daniel; Kai, Lei; Karbyshev, Mikhail; Loehr, Frank; Bernhard, Frank; Doetsch, Volker

    2010-01-01

    Membrane proteins are highly underrepresented in the structural data-base and remain one of the most challenging targets for functional and structural elucidation. Their roles in transport and cellular communication, furthermore, often make over-expression toxic to their host, and their hydrophobicity and structural complexity make isolation and reconstitution a complicated task, especially in cases where proteins are targeted to inclusion bodies. The development of cell-free expression systems provides a very interesting alternative to cell-based systems, since it circumvents many problems such as toxicity or necessity for the transportation of the synthesized protein to the membrane, and constitutes the only system that allows for direct production of membrane proteins in membrane-mimetic environments which may be suitable for liquid state NMR measurements. The unique advantages of the cell-free expression system, including strong expression yields as well as the direct incorporation of almost any combination of amino acids with very little metabolic scrambling, has allowed for the development of a wide-array of isotope labelling techniques which facilitate structural investigations of proteins whose spectral congestion and broad line-widths may have earlier rendered them beyond the scope of NMR. Here we explore various labelling strategies in conjunction with cell-free developments, with a particular focus on α-helical transmembrane proteins which benefit most from such methods.

  9. Processed Meat Protein and Heat-Stable Peptide Marker Identification Using Microwave-Assisted Tryptic Digestion

    Directory of Open Access Journals (Sweden)

    Magdalena Montowska

    2016-01-01

    Full Text Available New approaches to rapid examination of proteins and peptides in complex food matrices are of great interest to the community of food scientists. The aim of the study is to examine the influence of microwave irradiation on the acceleration of enzymatic cleavage and enzymatic digestion of denatured proteins in cooked meat of five species (cattle, horse, pig, chicken and turkey and processed meat products (coarsely minced, smoked, cooked and semi-dried sausages. Severe protein aggregation occurred not only in heated meat under harsh treatment at 190 °C but also in processed meat products. All the protein aggregates were thoroughly hydrolyzed aft er 1 h of trypsin treatment with short exposure times of 40 and 20 s to microwave irradiation at 138 and 303 W. There were much more missed cleavage sites observed in all microwave-assisted digestions. Despite the incompleteness of microwave-assisted digestion, six unique peptide markers were detected, which allowed unambiguous identification of processed meat derived from the examined species. Although the microwave-assisted tryptic digestion can serve as a tool for rapid and high-throughput protein identification, great caution and pre-evaluation of individual samples is recommended in protein quantitation.

  10. Concentration-Induced Association in a Protein System Caused by a Highly Directional Patch Attraction.

    Science.gov (United States)

    Li, Weimin; Persson, Björn A; Lund, Mikael; Bergenholtz, Johan; Zackrisson Oskolkova, Malin

    2016-09-01

    Self-association of the protein lactoferrin is studied in solution using small-angle X-ray scattering techniques. Effective static structure factors have been shown to exhibit either a monotonic or a nonmonotonic dependence on protein concentration in the small wavevector limit, depending on salt concentration. The behavior correlates with a nonmonotonic dependence of the second virial coefficient on salt concentration, such that a maximum appears in the structure factor at a low protein concentration when the second virial coefficient is negative and close to a minimum. The results are interpreted in terms of an integral equation theory with explicit dimers, formulated by Wertheim, which provides a consistent framework able to explain the behavior in terms of a monomer-dimer equilibrium that appears because of a highly directional patch attraction. Short attraction ranges preclude trimer formation, which explains why the protein system behaves as if it were subject to a concentration-dependent isotropic protein-protein attraction. Superimposing an isotropic interaction, comprising screened Coulomb repulsion and van der Waals attraction, on the patch attraction allows for a semiquantitative modeling of the complete transition pathway from monomers in the dilute limit to monomer-dimer systems at somewhat higher protein concentrations.

  11. High Concentration Protein Ultrafiltration: a Comparative Fouling Assessment

    Science.gov (United States)

    Lim, Y. P.; Mohammad, A. W.

    2018-05-01

    In this paper, the predominant fouling mechanism via pH manipulation in gelatin ultrafiltration (UF) at constant operating pressure was studied. Two 30 kDa molecular weight cut off (MWCO) UF membranes with different hydrophilic/hydrophobic properties were tested at solution pH near gelatin isoelectric point (IEP), pH below and above gelatin’s IEP. The resistance-in-series model was used to determine quantitatively the contribution of each filtration resistance occurred during gelatin UF. The governing fouling mechanisms were investigated using classical blocking laws. The results demonstrated that concentration polarization remain as dominant fouling resistance in gelatin UF, but exceptional case was observed at pH away from gelatin’s IEP, showing that combined reversible and irreversible fouling resistances contributed around 57% and 37%, respectively to the overall fouling resistances. Under all experimental condition tested, permeate flux decline was accurately predicted by all the models studied. Fouling profile was fitted well with “Standard Blocking”, “Intermediate Blocking” and “Cake Filtration” model for regenerated cellulose acetate (RCA) membrane and “Cake Filtration” model for polyethersulphone (PES) membrane.

  12. A stable aspirin-triggered lipoxin A4 analog blocks phosphorylation of leukocyte-specific protein 1 in human neutrophils.

    Science.gov (United States)

    Ohira, Taisuke; Bannenberg, Gerard; Arita, Makoto; Takahashi, Minoru; Ge, Qingyuan; Van Dyke, Thomas E; Stahl, Gregory L; Serhan, Charles N; Badwey, John A

    2004-08-01

    Lipoxins and their aspirin-triggered 15-epimers are endogenous anti-inflammatory agents that block neutrophil chemotaxis in vitro and inhibit neutrophil influx in several models of acute inflammation. In this study, we examined the effects of 15-epi-16-(p-fluoro)-phenoxy-lipoxin A(4) methyl ester, an aspirin-triggered lipoxin A(4)-stable analog (ATLa), on the protein phosphorylation pattern of human neutrophils. Neutrophils stimulated with the chemoattractant fMLP were found to exhibit intense phosphorylation of a 55-kDa protein that was blocked by ATLa (10-50 nM). This 55-kDa protein was identified as leukocyte-specific protein 1, a downstream component of the p38-MAPK cascade in neutrophils, by mass spectrometry, Western blotting, and immunoprecipitation experiments. ATLa (50 nM) also reduced phosphorylation/activation of several components of the p38-MAPK pathway in these cells (MAPK kinase 3/MAPK kinase 6, p38-MAPK, MAPK-activated protein kinase-2). These results indicate that ATLa exerts its anti-inflammatory effects, at least in part, by blocking activation of the p38-MAPK cascade in neutrophils, which is known to promote chemotaxis and other proinflammatory responses by these cells.

  13. CAG Expansions Are Genetically Stable and Form Nontoxic Aggregates in Cells Lacking Endogenous Polyglutamine Proteins

    Directory of Open Access Journals (Sweden)

    Ashley A. Zurawel

    2016-09-01

    Full Text Available Proteins containing polyglutamine (polyQ regions are found in almost all eukaryotes, albeit with various frequencies. In humans, proteins such as huntingtin (Htt with abnormally expanded polyQ regions cause neurodegenerative diseases such as Huntington’s disease (HD. To study how the presence of endogenous polyQ aggregation modulates polyQ aggregation and toxicity, we expressed polyQ expanded Htt fragments (polyQ Htt in Schizosaccharomyces pombe. In stark contrast to other unicellular fungi, such as Saccharomyces cerevisiae, S. pombe is uniquely devoid of proteins with more than 10 Q repeats. We found that polyQ Htt forms aggregates within S. pombe cells only with exceedingly long polyQ expansions. Surprisingly, despite the presence of polyQ Htt aggregates in both the cytoplasm and nucleus, no significant growth defect was observed in S. pombe cells. Further, PCR analysis showed that the repetitive polyQ-encoding DNA region remained constant following transformation and after multiple divisions in S. pombe, in contrast to the genetic instability of polyQ DNA sequences in other organisms. These results demonstrate that cells with a low content of polyQ or other aggregation-prone proteins can show a striking resilience with respect to polyQ toxicity and that genetic instability of repetitive DNA sequences may have played an important role in the evolutionary emergence and exclusion of polyQ expansion proteins in different organisms.

  14. Periodontal status affects C-reactive protein and lipids in patients with stable heart disease from a tertiary care cardiovascular clinic.

    Science.gov (United States)

    Flores, Manuela F; Montenegro, Marlon M; Furtado, Mariana V; Polanczyk, Carisi A; Rösing, Cassiano K; Haas, Alex N

    2014-04-01

    There are scarce data on the impact of the periodontal condition in the control of biomarkers in patients with cardiovascular disease (CVD). The aim of this study is to assess whether periodontal inflammation and tissue breakdown are associated with C-reactive protein (CRP) and lipids in patients with stable heart disease. This cross-sectional study included 93 patients with stable coronary artery disease (57 males; mean age: 63.5 ± 9.8 years) who were in outpatient care for at least 6 months. After applying a structured questionnaire, periodontal examinations were performed by two calibrated periodontists in six sites per tooth at all teeth. Blood samples were collected from patients on the day of periodontal examination to determine levels of CRP, lipids, and glycated hemoglobin. Multiple linear regression models were fitted to evaluate the association among different periodontal and blood parameters controlling for sex, body mass index, glycated hemoglobin, use of oral hypoglycemic drugs, and smoking. Overall, the sample presented high levels of periodontal inflammation and tissue breakdown. Unadjusted mean concentrations of triglycerides (TGs), very-low-density lipoprotein cholesterol, and glucose were significantly higher in individuals with severe periodontitis. When multiple linear regression models were applied, number of teeth with clinical attachment loss ≥6 mm and presence of severe periodontitis were significantly associated with higher CRP concentrations. Bleeding on probing was significantly associated with TGs, total cholesterol, and non-high-density lipoprotein cholesterol. In this sample of patients with stable CVD, current periodontal inflammation and tissue breakdown are associated with cardiovascular inflammatory markers, such as CRP and lipid profile.

  15. Randomly organized lipids and marginally stable proteins: a coupling of weak interactions to optimize membrane signaling.

    Science.gov (United States)

    Rice, Anne M; Mahling, Ryan; Fealey, Michael E; Rannikko, Anika; Dunleavy, Katie; Hendrickson, Troy; Lohese, K Jean; Kruggel, Spencer; Heiling, Hillary; Harren, Daniel; Sutton, R Bryan; Pastor, John; Hinderliter, Anne

    2014-09-01

    Eukaryotic lipids in a bilayer are dominated by weak cooperative interactions. These interactions impart highly dynamic and pliable properties to the membrane. C2 domain-containing proteins in the membrane also interact weakly and cooperatively giving rise to a high degree of conformational plasticity. We propose that this feature of weak energetics and plasticity shared by lipids and C2 domain-containing proteins enhance a cell's ability to transduce information across the membrane. We explored this hypothesis using information theory to assess the information storage capacity of model and mast cell membranes, as well as differential scanning calorimetry, carboxyfluorescein release assays, and tryptophan fluorescence to assess protein and membrane stability. The distribution of lipids in mast cell membranes encoded 5.6-5.8bits of information. More information resided in the acyl chains than the head groups and in the inner leaflet of the plasma membrane than the outer leaflet. When the lipid composition and information content of model membranes were varied, the associated C2 domains underwent large changes in stability and denaturation profile. The C2 domain-containing proteins are therefore acutely sensitive to the composition and information content of their associated lipids. Together, these findings suggest that the maximum flow of signaling information through the membrane and into the cell is optimized by the cooperation of near-random distributions of membrane lipids and proteins. This article is part of a Special Issue entitled: Interfacially Active Peptides and Proteins. Guest Editors: William C. Wimley and Kalina Hristova. Copyright © 2014 Elsevier B.V. All rights reserved.

  16. Effect of membrane protein concentration on binding of 3H-imipramine in human platelets

    International Nuclear Information System (INIS)

    Barkai, A.I.; Kowalik, S.; Baron, M.

    1985-01-01

    Binding of 3 H-imipramine to platelet membranes has been implicated as a marker for depression. Comparing 3 H-IMI binding between depressed patients and normal subjects we observed an increase in the dissociation constant Kd with increasing membrane protein. This phenomenon was studied more rigorously in five normal subjects. Platelet membranes were prepared and adjusted to four concentrations of protein ranging from 100 to 800 micrograms/ml. The 3 H-IMI binding parameters of maximum binding sites number (Bmax) and Kd were obtained by Scatchard analysis at each membrane concentration. A positive linear relationship was found between K/sub d/ values and the concentration of membrane protein in the assay, but no change was observed in Bmax. The variability in Kd values reported in the literature may be accounted for in part by the different concentrations of membrane protein used in various studies

  17. Naturally occurring stable isotopes reflect changes in protein turnover and growth in gilthead sea bream (Sparus aurata) juveniles under different dietary protein levels.

    Science.gov (United States)

    Martin-Perez, Miguel; Fernandez-Borras, Jaume; Ibarz, Antoni; Felip, Olga; Fontanillas, Ramon; Gutierrez, Joaquim; Blasco, Josefina

    2013-09-18

    Ideal nutritional conditions are crucial to sustainable aquaculture due to economic and environmental issues. Here we apply stable isotope analysis as an indicator of fish growth and feeding balance, to define the optimum diet for efficient growing conditions. Juveniles of gilthead sea bream were fed with six isoenergetic diets differing in protein to lipid proportion (from 41/26 to 57/20). As protein intake increased, δ¹⁵N and Δδ¹⁵N of muscle and Δδ¹⁵N and Δδ¹³C of its protein fraction decreased, indicating lower protein turnover and higher protein deposition in muscle. This is reflected in the inverse relationship found between Δδ¹⁵N and growth rate, although no differences were observed in either parameter beyond the protein/lipid proportion 47/23. Principal component analysis (PCA) also signaled 47/23 diet as the pivotal point with the highest growing efficiency, with isotopic parameters having the highest discrimination load. Thus, muscle isotope composition, especially ¹⁵N, can be used to evaluate nutritional status in farmed fish.

  18. An Improved Method of Predicting Extinction Coefficients for the Determination of Protein Concentration.

    Science.gov (United States)

    Hilario, Eric C; Stern, Alan; Wang, Charlie H; Vargas, Yenny W; Morgan, Charles J; Swartz, Trevor E; Patapoff, Thomas W

    2017-01-01

    Concentration determination is an important method of protein characterization required in the development of protein therapeutics. There are many known methods for determining the concentration of a protein solution, but the easiest to implement in a manufacturing setting is absorption spectroscopy in the ultraviolet region. For typical proteins composed of the standard amino acids, absorption at wavelengths near 280 nm is due to the three amino acid chromophores tryptophan, tyrosine, and phenylalanine in addition to a contribution from disulfide bonds. According to the Beer-Lambert law, absorbance is proportional to concentration and path length, with the proportionality constant being the extinction coefficient. Typically the extinction coefficient of proteins is experimentally determined by measuring a solution absorbance then experimentally determining the concentration, a measurement with some inherent variability depending on the method used. In this study, extinction coefficients were calculated based on the measured absorbance of model compounds of the four amino acid chromophores. These calculated values for an unfolded protein were then compared with an experimental concentration determination based on enzymatic digestion of proteins. The experimentally determined extinction coefficient for the native proteins was consistently found to be 1.05 times the calculated value for the unfolded proteins for a wide range of proteins with good accuracy and precision under well-controlled experimental conditions. The value of 1.05 times the calculated value was termed the predicted extinction coefficient. Statistical analysis shows that the differences between predicted and experimentally determined coefficients are scattered randomly, indicating no systematic bias between the values among the proteins measured. The predicted extinction coefficient was found to be accurate and not subject to the inherent variability of experimental methods. We propose the use of a

  19. Protein-free transfection of CHO host cells with an IgG-fusion protein: selection and characterization of stable high producers and comparison to conventionally transfected clones.

    Science.gov (United States)

    Lattenmayer, Christine; Loeschel, Martina; Schriebl, Kornelia; Steinfellner, Willibald; Sterovsky, Thomas; Trummer, Evelyn; Vorauer-Uhl, Karola; Müller, Dethardt; Katinger, Hermann; Kunert, Renate

    2007-04-15

    In order to improve the current techniques of cell cultivation in the absence of serum, we have developed a protein-free transfection protocol for CHO cells, based on the Nucleofector technology. After starting with a heterogeneous pool of primary transfectants which express the fusion protein EpoFc, we isolated single clones and compared them with parallel clones generated by lipofection in serum-dependent cultivation. Our intensive characterization program was based on determination of specific productivity (q(p)) and analysis of genetic parameters. In two nucleofection experiments, transfection with 5 microg of DNA resulted in best productivities of the primary cell pools. After subcloning, the q(p) could be raised up to 27 pg x cells(-1) x day(-1). While the serum-dependent transfectants exhibited specific productivities up to 57 pg x cells(-1) x day(-1) in serum-dependent cultivation, a significant decrease that resulted in the range of q(p) of the protein-free transfectants was observed after switching to protein-free conditions. Investigation of genetic parameters revealed higher mRNA levels and gene copy numbers (GCN) for the protein-free adapted serum-dependent transfectants. Therefore, we assume that problems during protein-free adaptation (PFA) lead to a less efficient translation machinery after serum deprivation. We describe the generation of stable-producing recombinant CHO clones by protein-free transfection of a protein-free adapted host cell line, which reduces the risk of adverse clonal changes after PFA. The main advantage of this approach is the earlier predictability of clone behavior, which makes the generation of production clones by protein-free transfection, a viable and highly efficient strategy for recombinant cell line development. (c) 2006 Wiley Periodicals, Inc.

  20. Pregnancy associated plasma protein-A as a marker for myocardial infarction and death in patients with stable coronary artery disease: A prognostic study within the CLARICOR Trial

    DEFF Research Database (Denmark)

    Iversen, Kasper K; Teisner, Børge; Winkel, Per

    2011-01-01

    OBJECTIVE: Pregnancy associated plasma protein-A (PAPP-A) is a potential new marker for vulnerable plaques in the coronary arteries only examined in stable coronary disease (CAD) in patients undergoing coronary angiography. Here we address the prognostic value of serum PAPP-A in unselected stable...

  1. Stable Plastid Transformation for High-Level Recombinant Protein Expression: Promises and Challenges

    Directory of Open Access Journals (Sweden)

    Meili Gao

    2012-01-01

    Full Text Available Plants are a promising expression system for the production of recombinant proteins. However, low protein productivity remains a major obstacle that limits extensive commercialization of whole plant and plant cell bioproduction platform. Plastid genetic engineering offers several advantages, including high levels of transgenic expression, transgenic containment via maternal inheritance, and multigene expression in a single transformation event. In recent years, the development of optimized expression strategies has given a huge boost to the exploitation of plastids in molecular farming. The driving forces behind the high expression level of plastid bioreactors include codon optimization, promoters and UTRs, genotypic modifications, endogenous enhancer and regulatory elements, posttranslational modification, and proteolysis. Exciting progress of the high expression level has been made with the plastid-based production of two particularly important classes of pharmaceuticals: vaccine antigens, therapeutic proteins, and antibiotics and enzymes. Approaches to overcome and solve the associated challenges of this culture system that include low transformation frequencies, the formation of inclusion bodies, and purification of recombinant proteins will also be discussed.

  2. Legume Protein Isolates for Stable Acidic Emulsions Prepared by Premix Membrane Emulsification

    NARCIS (Netherlands)

    Ladjal Ettoumi, Yakoub; Berton-Carabin, Claire; Chibane, Mohamed; Schroën, Karin

    2017-01-01

    Proteins originating from dry legumes are not that much used in food formulations, yet, they are interesting components from a sustainability point of view, and could have interesting functional properties, e.g. for emulsion preparation. Therefore, this work focuses on the potential of the water

  3. Residues of the UL25 Protein of Herpes Simplex Virus That Are Required for Its Stable Interaction with Capsids ▿

    Science.gov (United States)

    Cockrell, Shelley K.; Huffman, Jamie B.; Toropova, Katerina; Conway, James F.; Homa, Fred L.

    2011-01-01

    The herpes simplex virus 1 (HSV-1) UL25 gene product is a minor capsid component that is required for encapsidation, but not cleavage, of replicated viral DNA. UL25 is located on the capsid surface in a proposed heterodimer with UL17, where five copies of the heterodimer are found at each of the capsid vertices. Previously, we demonstrated that amino acids 1 to 50 of UL25 are essential for its stable interaction with capsids. To further define the UL25 capsid binding domain, we generated recombinant viruses with either small truncations or amino acid substitutions in the UL25 N terminus. Studies of these mutants demonstrated that there are two important regions within the capsid binding domain. The first 27 amino acids are essential for capsid binding of UL25, while residues 26 to 39, which are highly conserved in the UL25 homologues of other alphaherpesviruses, were found to be critical for stable capsid binding. Cryo-electron microscopy reconstructions of capsids containing either a small tag on the N terminus of UL25 or the green fluorescent protein (GFP) fused between amino acids 50 and 51 of UL25 demonstrate that residues 1 to 27 of UL25 contact the hexon adjacent to the penton. A second region, most likely centered on amino acids 26 to 39, contacts the triplex that is one removed from the penton. Importantly, both of these UL25 capsid binding regions are essential for the stable packaging of full-length viral genomes. PMID:21411517

  4. Effects of Ionic Strength on the Enzymatic Hydrolysis of Diluted and Concentrated Whey Protein Isolate

    NARCIS (Netherlands)

    Butré, C.I.; Wierenga, P.A.; Gruppen, H.

    2012-01-01

    To identify the parameters that affect enzymatic hydrolysis at high substrate concentrations, whey protein isolate (1–30% w/v) was hydrolyzed by Alcalase and Neutrase at constant enzyme-to-substrate ratio. No changes were observed in the solubility and the aggregation state of the proteins. With

  5. Protein Concentration in Milk Formula, Growth, and Later Risk of Obesity: A Systematic Review

    NARCIS (Netherlands)

    Patro-Gołąb, Bernadeta; Zalewski, Bartłomiej M.; Kouwenhoven, Stefanie M. P.; Karaś, Jacek; Koletzko, Berthold; van Goudoever, Johannes Bernard; Szajewska, Hania

    2016-01-01

    Background: Protein intake may influence important health outcomes in later life. Objective: The objective of this study was to investigate current evidence on the effects of infant formulas and follow-on formulas with different protein concentrations on infants' and children's growth, body

  6. Concentration of Proteins and Protein Fractions in Blood Plasma of Chickens Hatched from Eggs Irradiated with Low Level Gamma Rays

    International Nuclear Information System (INIS)

    Kraljevic, P.; Vilic, M.; Simpraga, M.; Matisic, D.; Miljanic, S.

    2011-01-01

    In literature there are many results which have shown that low dose radiation can stimulate many physiological processes of living organism. In our earlier paper it was shown that low dose of gamma radiation has a stimulative effect upon metabolic process in chickens hatched from eggs irradiated before incubation. This was proved by increase of body weight gain and body weight, as well as by increase of two enzymes activities in blood plasma (aspartate aminotransferase and alanine aminotransferase) which play an important role in protein metabolism. Therefore, an attempt was made to determine the effect of eggs irradiation by low dose gamma rays upon concentration of total proteins and protein fractions in the blood plasma of chickens hatched from irradiated eggs. The eggs of heavy breed chickens were irradiated with a dose of 0.15 Gy gamma radiation (60Co) before incubation. Along with the chickens which were hatched from irradiated eggs, there was a control group of chickens hatched from nonirradiated eggs. All other conditions were the same for both groups of chickens. Blood samples were taken from the right jugular vein on the 1 s t and 3 r d day, or from the wing vein on days 5 and 7 after hatching. The total proteins concentration in the blood plasma was determined by the biuret method using Boehringer Mannheim GmbH optimized kits. The protein fractions (albumin, α 1 -globulin, α 2 -globulin, β- and γ-globulins) were estimated electrophoretically on Cellogel strips. The total proteins concentration was significantly decreased in blood plasma of chickens hatched from irradiated eggs on days 3 (P t h day (P 2 -globulin was decreased on days 1 (P t h day of life. Obtained results indicate that low dose of gamma radiation has mostly inhibitory effect upon concentration of total proteins and protein fractions in the blood plasma of chickens hatched from irradiated eggs before incubation. (author)

  7. Cold-set globular protein gels: Interactions, structure and rheology as a function of protein concentration.

    NARCIS (Netherlands)

    Alting, A.C.; Hamer, R.J.; Kruif, de C.G.

    2003-01-01

    We identified the contribution of covalent and noncovalent interactions to the scaling behavior of the structural and rheological properties in a cold gelling protein system. The system we studied consisted of two types of whey protein aggregates, equal in size but different in the amount of

  8. A novel approach for the detection of potentially hazardous pepsin stable hazelnut proteins as contaminants in chocolate-based food.

    Science.gov (United States)

    Akkerdaas, Jaap H; Wensing, Marjolein; Knulst, André C; Stephan, Oliver; Hefle, Susan L; Aalberse, Rob C; van Ree, Ronald

    2004-12-15

    Contamination of food products with pepsin resistant allergens is generally believed to be a serious threat to patients with severe food allergy. A sandwich type enzyme-linked immunosorbent assay (ELISA) was developed to measure pepsin resistant hazelnut protein in food products. Capturing and detecting rabbit antibodies were raised against pepsin-digested hazelnut and untreated hazelnut protein, respectively. The assay showed a detection limit of 0.7 ng/mL hazelnut protein or food matrix and a maximum of 0.034% cross-reactivity (peanut). Chocolate samples spiked with 0.5-100 microg hazelnut/g chocolate showed a mean recovery of 97.3%. In 9/12 food products labeled "may contain nuts", hazelnut was detected between 1.2 and 417 microg hazelnut/g food. It can be concluded that the application of antibodies directed to pepsin-digested food extracts in ELISA can facilitate specific detection of stable proteins that have the highest potential of inducing severe food anaphylaxis.

  9. Recombination-stable multimeric green fluorescent protein for characterization of weak promoter outputs in Saccharomyces cerevisiae

    DEFF Research Database (Denmark)

    Rugbjerg, Peter; Knuf, Christoph; Förster, Jochen

    2015-01-01

    a less leaky Cu2+-inducible promoter based on CUP1. The basal expression level of the new promoter was approx. 61% below the wild-type CUP1 promoter, thus expanding the absolute range of Cu2+-based gene control. The stability of 3vGFP towards direct-repeat recombination was assayed in S. cerevisiae......Green fluorescent proteins (GFPs) are widely used for visualization of proteins to track localization and expression dynamics. However, phenotypically important processes can operate at too low expression levels for routine detection, i.e. be overshadowed by autofluorescence noise. While GFP...... functions well in translational fusions, the use of tandem GFPs to amplify fluorescence signals is currently avoided in Saccharomyces cerevisiae and many other microorganisms due to the risk of loop-out by direct-repeat recombination. We increased GFP fluorescence by translationally fusing three different...

  10. Structural changes induced by high-pressure processing in micellar casein and milk protein concentrates.

    Science.gov (United States)

    Cadesky, Lee; Walkling-Ribeiro, Markus; Kriner, Kyle T; Karwe, Mukund V; Moraru, Carmen I

    2017-09-01

    Reconstituted micellar casein concentrates and milk protein concentrates of 2.5 and 10% (wt/vol) protein concentration were subjected to high-pressure processing at pressures from 150 to 450 MPa, for 15 min, at ambient temperature. The structural changes induced in milk proteins by high-pressure processing were investigated using a range of physical, physicochemical, and chemical methods, including dynamic light scattering, rheology, mid-infrared spectroscopy, scanning electron microscopy, proteomics, and soluble mineral analyses. The experimental data clearly indicate pressure-induced changes of casein micelles, as well as denaturation of serum proteins. Calcium-binding α S1 - and α S2 -casein levels increased in the soluble phase after all pressure treatments. Pressurization up to 350 MPa also increased levels of soluble calcium and phosphorus, in all samples and concentrations, whereas treatment at 450 MPa reduced the levels of soluble Ca and P. Experimental data suggest dissociation of calcium phosphate and subsequent casein micelle destabilization as a result of pressure treatment. Treatment of 10% micellar casein concentrate and 10% milk protein concentrate samples at 450 MPa resulted in weak, physical gels, which featured aggregates of uniformly distributed, casein substructures of 15 to 20 nm in diameter. Serum proteins were significantly denatured by pressures above 250 MPa. These results provide information on pressure-induced changes in high-concentration protein systems, and may inform the development on new milk protein-based foods with novel textures and potentially high nutritional quality, of particular interest being the soft gel structures formed at high pressure levels. The Authors. Published by the Federation of Animal Science Societies and Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/).

  11. Alkali solution extraction of rice residue protein isolates: Influence of alkali concentration on protein functional, structural properties and lysinoalanine formation.

    Science.gov (United States)

    Hou, Furong; Ding, Wenhui; Qu, Wenjuan; Oladejo, Ayobami Olayemi; Xiong, Feng; Zhang, Weiwei; He, Ronghai; Ma, Haile

    2017-03-01

    This study evaluated the nutrient property and safety of the rice residue protein isolates (RRPI) product (extracted by different alkali concentrations) by exploring the protein functional, structural properties and lysinoalanine (LAL) formation. The results showed that with the rising of alkali concentration from 0.03M to 0.15M, the solubility, emulsifying and foaming properties of RRPI increased at first and then descended. When the alkali concentration was greater than 0.03M, the RRPI surface hydrophobicity decreased and the content of thiol and disulfide bond, Lys and Cys significantly reduced. By the analysis of HPLC, the content of LAL rose up from 276.08 to 15,198.07mg/kg and decreased to 1340.98mg/kg crude protein when the alkali concentration increased from 0.03 to 0.09M and until to 0.15M. These results indicated that RRPI alkaline extraction concentration above 0.03M may cause severe nutrient or safety problems of protein. Copyright © 2016 Elsevier Ltd. All rights reserved.

  12. Semen quality and concentration of soluble proteins in the seminal plasma of Alpine bucks Semen quality and concentration of soluble proteins in the seminal plasma of Alpine bucks

    Directory of Open Access Journals (Sweden)

    Simone Eliza Facione Guimarães

    2010-06-01

    Full Text Available It was aimed to study the in vitro seminal quality analyzed by complementary tests and to compare them with physical, morphological and biochemical aspects of male goat semen of the Alpine breed. This experiment took place at the Federal University of Viçosa, situated at 20º45’ S latitude and 42º51’ W longitude, Southwest of Brazil. It was done during the summer months of January and February, and three adult male goats of the Alpine breed were used in intensive conditions. The semen was collected by artificial vagina method. In all semen samples (45 ejaculates, after the physical and morphological analysis, the hiposmotic test was done. In 24 ejaculates, it were done thermo-resistance test, and in 21 ejaculates it were determined the concentration of total soluble proteins in seminal plasma. The male goats presented difference in the semen physical and morphological aspects, in the hiposmotic test and thermo-resistance test, but they did not presented difference in total soluble proteins concentration in seminal plasma. Results of the slow thermo-resistance test and hiposmotic test were positively correlated (r = 0.60. It was concluded, according to our results, that the concentration of total soluble proteins in seminal plasma can not be used as a parameter to predict the seminal quality of Alpine bucks.It was aimed to study the in vitro seminal quality analyzed by complementary tests and to compare them with physical, morphological and biochemical aspects of male goat semen of the Alpine breed. This experiment took place at the Federal University of Viçosa, situated at 20º45’ S latitude and 42º51’ W longitude, Southwest of Brazil. It was done during the summer months of January and February, and three adult male goats of the Alpine breed were used in intensive conditions. The semen was collected by artificial vagina method. In all semen samples (45 ejaculates, after the physical and morphological analysis, the hiposmotic test

  13. A stable and convenient protein electrophoresis titration device with bubble removing system.

    Science.gov (United States)

    Zhang, Qiang; Fan, Liu-Yin; Li, Wen-Lin; Cong, Feng-Song; Zhong, Ran; Chen, Jing-Jing; He, Yu-Chen; Xiao, Hua; Cao, Cheng-Xi

    2017-07-01

    Moving reaction boundary titration (MRBT) has a potential application to immunoassay and protein content analysis with high selectivity. However, air bubbles often impair the accuracy of MRBT, and the leakage of electrolyte greatly decreases the safety and convenience of electrophoretic titration. Addressing these two issues a reliable MRBT device with modified electrolyte chamber of protein titration was designed. Multiphysics computer simulation was conducted for optimization according to two-phase flow. The single chamber was made of two perpendicular cylinders with different diameters. After placing electrophoretic tube, the resident air in the junction next to the gel could be eliminated by a simple fast electrolyte flow. Removing the electrophoretic tube automatically prevented electrolyte leakage at the junction due to the gravity-induced negative pressure within the chamber. Moreover, the numerical simulation and experiments showed that the improved MRBT device has following advantages: (i) easy and rapid setup of electrophoretic tube within 20 s; (ii) simple and quick bubble dissipates from the chamber of titration within 2 s; (iii) no electrolyte leakage from the two chambers: and (iv) accurate protein titration and safe instrumental operation. The developed technique and apparatus greatly improves the performance of the previous MRBT device, and providing a new route toward practical application. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  14. Milk protein concentration, estimated breeding value for fertility, and reproductive performance in lactating dairy cows.

    Science.gov (United States)

    Morton, John M; Auldist, Martin J; Douglas, Meaghan L; Macmillan, Keith L

    2017-07-01

    Milk protein concentration in dairy cows has been positively associated with a range of measures of reproductive performance, and genetic factors affecting both milk protein concentration and reproductive performance may contribute to the observed phenotypic associations. It was of interest to assess whether these beneficial phenotypic associations are accounted for or interact with the effects of estimated breeding values for fertility. The effects of a multitrait estimated breeding value for fertility [the Australian breeding value for daughter fertility (ABV fertility)] on reproductive performance were also of interest. Interactions of milk protein concentration and ABV fertility with the interval from calving date to the start of the herd's seasonally concentrated breeding period were also assessed. A retrospective single cohort study was conducted using data collected from 74 Australian seasonally and split calving dairy herds. Associations between milk protein concentration, ABV fertility, and reproductive performance in Holstein cows were assessed using random effects logistic regression. Between 52,438 and 61,939 lactations were used for analyses of 4 reproductive performance measures. Milk protein concentration was strongly and positively associated with reproductive performance in dairy cows, and this effect was not accounted for by the effects of ABV fertility. Increases in ABV fertility had important additional beneficial effects on the probability of pregnancy by wk 6 and 21 of the herd's breeding period. For cows calved before the start of the breeding period, the effects of increases in both milk protein concentration and ABV fertility were beneficial regardless of their interval from calving to the start of the breeding period. These findings demonstrate the potential for increasing reproductive performance through identifying the causes of the association between milk protein concentration and reproductive performance and then devising management

  15. Measurement of Hepatic Protein Fractional Synthetic Rate with Stable Isotope Labeling Technique in Thapsigargin Stressed HepG2 Cells

    Science.gov (United States)

    Song, Juquan; Zhang, Xiao-jun; Boehning, Darren; Brooks, Natasha C.; Herndon, David N.; Jeschke, Marc G.

    2012-01-01

    Severe burn-induced liver damage and dysfunction is associated with endoplasmic reticulum (ER) stress. ER stress has been shown to regulate global protein synthesis. In the current study, we induced ER stress in vitro and estimated the effect of ER stress on hepatic protein synthesis. The aim was two-fold: (1) to establish an in vitro model to isotopically measure hepatic protein synthesis and (2) to evaluate protein fractional synthetic rate (FSR) in response to ER stress. Human hepatocellular carcinoma cells (HepG2) were cultured in medium supplemented with stable isotopes 1,2-13C2-glycine and L-[ring-13C6]phenylalanine. ER stress was induced by exposing the cells to 100 nM of thapsigargin (TG). Cell content was collected from day 0 to 14. Alterations in cytosolic calcium were measured by calcium imaging and ER stress markers were confirmed by Western blotting. The precursor and product enrichments were detected by GC-MS analysis for FSR calculation. We found that the hepatic protein FSR were 0.97±0.02 and 0.99±0.05%/hr calculated from 1,2-13C2-glycine and L-[ring-13C6]phenylalanine, respectively. TG depleted ER calcium stores and induced ER stress by upregulating p-IRE-1 and Bip. FSR dramatically decreased to 0.68±0.03 and 0.60±0.06%/hr in the TG treatment group (pisotope tracer incorporation technique is a useful method for studying the effects of ER stress on hepatic protein synthesis. PMID:22298954

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

    NARCIS (Netherlands)

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

    2015-01-01

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

  17. A Rapid Method for Determining the Concentration of Recombinant Protein Secreted from Pichia pastoris

    International Nuclear Information System (INIS)

    Sun, L W; Zhao, Y; Jiang, R; Song, Y; Feng, H; Feng, K; Niu, L P; Qi, C

    2011-01-01

    Pichia secretive expression system is one of powerful eukaryotic expression systems in genetic engineering, which is especially suitable for industrial utilization. Because of the low concentration of the target protein in initial experiment, the methods and conditions for expression of the target protein should be optimized according to the protein yield repetitively. It is necessary to set up a rapid, simple and convenient analysis method for protein expression levels instead of the generally used method such as ultrafiltration, purification, dialysis, lyophilization and so on. In this paper, acetone precipitation method was chosen to concentrate the recombinant protein firstly after comparing with four different protein precipitation methods systematically, and then the protein was analyzed by SDS-Polyacrylamide Gel Electrophoresis. The recombinant protein was determined with the feature of protein band by the Automated Image Capture and 1-D Analysis Software directly. With this method, the optimized expression conditions of basic fibroblast growth factor secreted from pichia were obtained, which is as the same as using traditional methods. Hence, a convenient tool to determine the optimized conditions for the expression of recombinant proteins in Pichia was established.

  18. The effect of dietary protein on reproduction in the mare. VI. Serum progestagen concentrations during pregnancy

    Directory of Open Access Journals (Sweden)

    F.E. Van Niekerk

    1998-07-01

    Full Text Available Sixty-four Thoroughbred and Anglo-Arab mares aged 6-12 years were used, of which 40 were non-lactating and 24 lactating. Foals from these 24 mares were weaned at the age of 6 months. Non-lactating and lactating mares were divided into 4 dietary groups each. The total daily protein intake and the protein quality (essential amino-acid content differed in the 4 groups of non-lactating and 4 groups of lactating mares. The mares were covered and the effect of the quantity and quality of dietary protein on serum progestagen concentrations during pregnancy was studied. A sharp decline in serum progestagen concentrations was recorded in all dietary groups from Days 18 to 40 of pregnancy, with some individual mares reaching values of less than 4 ng/mℓ. Serum progestagen concentrations recorded in some of the non-lactating mares on the low-quality protein diet increased to higher values (p<0.05 than those of mares in the other 3 dietary groups at 35-140 days of pregnancy. A similar trend was observed for the lactating mares on a low-quality protein diet at 30-84 days of pregnancy. No such trends were observed in any of the other dietary groups. High-quality protein supplementation increased serum progestagen concentrations during the 1st 30 days of pregnancy. Lactation depressed serum progestagen concentrations until after the foals were weaned.

  19. CONCENTRATION AND RECOVERY OF PROTEIN FROM TUNA COOKING JUICE BY FORWARD OSMOSIS

    Directory of Open Access Journals (Sweden)

    KHONGNAKORN W.

    2016-07-01

    Full Text Available Tuna cooking processing plants generate large amount of cooking juice containing a significant content of protein. Recovery and concentrating process of this valuable compound together with a low energy consumption process are of interest regarding full utilization concept and green process approach. Forward osmosis (FO was employed in this work to recover and concentrate tuna cooking juice. FO process could increase the protein concentration up to 9% with an average permeate flux of 2.54 L/m2h. The permeate flux however tended to decrease as protein concentration increased due to the impact of osmotic pressure of the feed and fouling on the membrane surface. Since tuna cooking juice consists of protein and minerals, membrane analyses indicated that fouling was more severe compared to the fouling caused by standard bovine serum albumin pure protein. However, the presence of minerals rendered it a quicker and lower energy process by comparison. These results indicated that FO is a promising technique in the recovery and concentration of tuna cooking juice protein.

  20. Influence of watermelon seed protein concentrates on dough handling, textural and sensory properties of cookies.

    Science.gov (United States)

    Wani, Ali Abas; Sogi, D S; Singh, Preeti; Khatkar, B S

    2015-04-01

    Fruit processing wastes contain numerous by products of potential use in food & allied industry. Watermelon seeds represent a major by-product of the processing waste and contain high amount of nutritional proteins. Protein rich cereal based products are in demand due to their health promoting benefits. With this aim, wheat flour was fortified with watermelon seed protein concentrates (2.5 %, 5 %, 7.5 % and 10 % levels) to prepare cookies with desirable physical, nutritional, and textural and sensory properties. Substitution levels of 5 % and 10 % significantly (p ≤ 0.05) increased the dough stability and mixing tolerance index, however pasting properties and dough extensibility decreased considerably above 5 % substitution levels. Cookie fracture force (kg) increased significantly (p ≤ 0.05) above 5 % fortification levels. Cookie spread factor (W/T) increased from 2.5 % to 7.5 % fortification levels, further increase showed negative impact. Sensory scores of the cookies showed that protein concentrate may be added up to 7.5 % fortification levels. This study revealed that watermelon protein concentrates can be fortified with protein concentrates upto 5-7.5 % levels in cookies to improve their protein quality.

  1. Optimization of mold wheat bread fortified with soy flour, pea flour and whey protein concentrate.

    Science.gov (United States)

    Erben, Melina; Osella, Carlos A

    2017-07-01

    The objective of this work was to study the effect of replacing a selected wheat flour for defatted soy flour, pea flour and whey protein concentrate on both dough rheological characteristics and the performance and nutritional quality of bread. A mixture design was used to analyze the combination of the ingredients. The optimization process suggested that a mixture containing 88.8% of wheat flour, 8.2% of defatted soy flour, 0.0% of pea flour and 3.0% of whey protein concentrate could be a good combination to achieve the best fortified-bread nutritional quality. The fortified bread resulted in high protein concentration, with an increase in dietary fiber content and higher calcium levels compared with those of control (wheat flour 100%). Regarding protein quality, available lysine content was significantly higher, thus contributing with the essential amino acid requirement.

  2. Increase in local protein concentration by field-inversion gel electrophoresis

    Directory of Open Access Journals (Sweden)

    Paulus Aran

    2007-09-01

    Full Text Available Abstract Background Proteins that migrate through cross-linked polyacrylamide gels (PAGs under the influence of a constant electric field experience negative factors, such as diffusion and non-specific trapping in the gel matrix. These negative factors reduce protein concentrations within a defined gel volume with increasing migration distance and, therefore, decrease protein separation efficiency. Enhancement of protein separation efficiency was investigated by implementing pulsed field-inversion gel electrophoresis (FIGE. Results Separation of model protein species and large protein complexes was compared between FIGE and constant field electrophoresis (CFE in different percentages of PAGs. Band intensities of proteins in FIGE with appropriate ratios of forward and backward pulse times were superior to CFE despite longer running times. These results revealed an increase in band intensity per defined gel volume. A biphasic protein relative mobility shift was observed in percentages of PAGs up to 14%. However, the effect of FIGE on protein separation was stochastic at higher PAG percentage. Rat liver lysates subjected to FIGE in the second-dimension separation of two-dimensional polyarcylamide gel electrophoresis (2D PAGE showed a 20% increase in the number of discernible spots compared with CFE. Nine common spots from both FIGE and CFE were selected for peptide sequencing by mass spectrometry (MS, which revealed higher final ion scores of all nine protein spots from FIGE. Native protein complexes ranging from 800 kDa to larger than 2000 kDa became apparent using FIGE compared with CFE. Conclusion The present investigation suggests that FIGE under appropriate conditions improves protein separation efficiency during PAGE as a result of increased local protein concentration. FIGE can be implemented with minimal additional instrumentation in any laboratory setting. Despite the tradeoff of longer running times, FIGE can be a powerful protein

  3. Increase in local protein concentration by field-inversion gel electrophoresis.

    Science.gov (United States)

    Tsai, Henghang; Low, Teck Yew; Freeby, Steve; Paulus, Aran; Ramnarayanan, Kalpana; Cheng, Chung-Pui Paul; Leung, Hon-Chiu Eastwood

    2007-09-26

    Proteins that migrate through cross-linked polyacrylamide gels (PAGs) under the influence of a constant electric field experience negative factors, such as diffusion and non-specific trapping in the gel matrix. These negative factors reduce protein concentrations within a defined gel volume with increasing migration distance and, therefore, decrease protein separation efficiency. Enhancement of protein separation efficiency was investigated by implementing pulsed field-inversion gel electrophoresis (FIGE). Separation of model protein species and large protein complexes was compared between FIGE and constant field electrophoresis (CFE) in different percentages of PAGs. Band intensities of proteins in FIGE with appropriate ratios of forward and backward pulse times were superior to CFE despite longer running times. These results revealed an increase in band intensity per defined gel volume. A biphasic protein relative mobility shift was observed in percentages of PAGs up to 14%. However, the effect of FIGE on protein separation was stochastic at higher PAG percentage. Rat liver lysates subjected to FIGE in the second-dimension separation of two-dimensional polyarcylamide gel electrophoresis (2D PAGE) showed a 20% increase in the number of discernible spots compared with CFE. Nine common spots from both FIGE and CFE were selected for peptide sequencing by mass spectrometry (MS), which revealed higher final ion scores of all nine protein spots from FIGE. Native protein complexes ranging from 800 kDa to larger than 2000 kDa became apparent using FIGE compared with CFE. The present investigation suggests that FIGE under appropriate conditions improves protein separation efficiency during PAGE as a result of increased local protein concentration. FIGE can be implemented with minimal additional instrumentation in any laboratory setting. Despite the tradeoff of longer running times, FIGE can be a powerful protein separation tool.

  4. Radioiododestannylation. Convenient synthesis of a stable penicillin derivative for rapid penicillin binding protein (PBP) assay

    International Nuclear Information System (INIS)

    Blaszczak, L.C.; Halligan, N.G.; Seitz, D.E.

    1989-01-01

    Radioiodination of p-(trimethylstannyl)penicillin V with [ 125 I]Na using a modification of the chloramine-T method is simple, high yielding, and site-specific. The structure and penicillin binding protein (PBP) affinity of p-[ 125 I]-penicillin V (IPV) are similar to penicillin G and the product can be used directly without purification in the PBP assay. Because of the high degree of stability toward autoradiolysis and equivalent PBP binding affinity, IPV can be used in place of [ 3 H]-penicillin G or [ 14 C]-penicillin G for these experiments. (author)

  5. Photonic reagents for concentration measurement of flu-orescent proteins with overlapping spectra

    Science.gov (United States)

    Goun, Alexei; Bondar, Denys I.; Er, Ali O.; Quine, Zachary; Rabitz, Herschel A.

    2016-05-01

    By exploiting photonic reagents (i.e., coherent control by shaped laser pulses), we employ Optimal Dynamic Discrimination (ODD) as a novel means for quantitatively characterizing mixtures of fluorescent proteins with a large spectral overlap. To illustrate ODD, we simultaneously measured concentrations of in vitro mixtures of Enhanced Blue Fluorescent Protein (EBFP) and Enhanced Cyan Fluorescent Protein (ECFP). Building on this foundational study, the ultimate goal is to exploit the capabilities of ODD for parallel monitoring of genetic and protein circuits by suppressing the spectral cross-talk among multiple fluorescent reporters.

  6. Determination of possible effects of mineral concentration on protein synthesis by rumen microbes in vitro

    International Nuclear Information System (INIS)

    Nikolic, J.A.; Jovanovic, M.; Andric, R.

    1976-01-01

    The aim of the present investigation was to determine the effect of different concentrations of sulphide, magnesium and zinc on protein synthesis by rumen micro-organisms in vitro. Rumen content was taken from a young bull fed a diet based on maize and dried sugar beet pulp (2/1) supplemented with urea. The rate of incorporation of 35 S from Na 2 35 SO 4 in relation to the mean specific radioactivity of the sulphide pool was used to estimate the overall rate of microbial protein synthesis. It was found that the rate of protein synthesis and the net rate of utilization of ammonia-N were not affected by differences in mean sulphide concentration from 3.6-8.0 mg/litre. The rate of reduction of sulphate appeared not to be affected by the addition of sodium sulphide to the medium. The rate and efficiency of protein synthesis by rumen micro-organisms were not significantly affected by increasing the concentration of total magnesium from 8.4-15.3 mg/100 ml. The values for soluble magnesium varied widely (1.2-7.8 mg/100 ml), and appeared to be partly dependent on the pH of the medium. Zinc concentrations varying from 5.2-12.4 mg/litre did not influence the overall rate of protein synthesis, although the efficiency tended to be higher when the concentration of zinc was greater. Concentrations of soluble zinc were low (0.3-1.15 mg/litre), and not influenced by changes in the concentration of total zinc. It was concluded that increasing the concentrations of the examined elements above the basic values did not lead consistently to an improved production of microbial protein but, on the other hand, had no obvious detrimental effect on microbial metabolic activity within the limits studied. (author)

  7. Stable megadalton TOC-TIC supercomplexes as major mediators of protein import into chloroplasts.

    Science.gov (United States)

    Chen, Lih-Jen; Li, Hsou-Min

    2017-10-01

    Preproteins are believed to be imported into chloroplasts through membrane contact sites where the translocon complexes of the outer (TOC) and inner (TIC) envelope membranes are assembled together. However, a single TOC-TIC supercomplex containing preproteins undergoing active import has not yet been directly observed. We optimized the blue native polyacrylamide gel electrophoresis (PAGE) (BN-PAGE) system to detect and resolve megadalton (MD)-sized complexes. Using this optimized system, the outer-membrane channel Toc75 from pea chloroplasts was found in at least two complexes: the 880-kD TOC complex and a previously undetected 1-MD complex. Two-dimensional BN-PAGE immunoblots further showed that Toc75, Toc159, Toc34, Tic20, Tic56 and Tic110 were all located in the 880-kD to 1.3-MD region. During active preprotein import, preproteins were transported mostly through the 1-MD complex and a smaller amount of preproteins was also detected in a complex of 1.25 MD. Antibody-shift assays showed that the 1-MD complex is a TOC-TIC supercomplex containing at least Toc75, Toc159, Toc34 and Tic110. Results from crosslinking and import with Arabidopsis chloroplasts suggest that the 1.25-MD complex is also a supercomplex. Our data provide direct evidence supporting that chloroplast preproteins are imported through TOC-TIC supercomplexes, and also provide the first size estimation of these supercomplexes. Furthermore, unlike in mitochondria where translocon supercomplexes are only transiently assembled during preprotein import, in chloroplasts at least some of the supercomplexes are preassembled stable structures. © 2017 The Authors The Plant Journal published by John Wiley & Sons Ltd and Society for Experimental Biology.

  8. Identification of syntrophic acetate-oxidizing bacteria in anaerobic digesters by combined protein-based stable isotope probing and metagenomics.

    Science.gov (United States)

    Mosbæk, Freya; Kjeldal, Henrik; Mulat, Daniel G; Albertsen, Mads; Ward, Alastair J; Feilberg, Anders; Nielsen, Jeppe L

    2016-10-01

    Inhibition of anaerobic digestion through accumulation of volatile fatty acids occasionally occurs as the result of unbalanced growth between acidogenic bacteria and methanogens. A fast recovery is a prerequisite for establishing an economical production of biogas. However, very little is known about the microorganisms facilitating this recovery. In this study, we investigated the organisms involved by a novel approach of mapping protein-stable isotope probing (protein-SIP) onto a binned metagenome. Under simulation of acetate accumulation conditions, formations of (13)C-labeled CO2 and CH4 were detected immediately following incubation with [U-(13)C]acetate, indicating high turnover rate of acetate. The identified (13)C-labeled peptides were mapped onto a binned metagenome for improved identification of the organisms involved. The results revealed that Methanosarcina and Methanoculleus were actively involved in acetate turnover, as were five subspecies of Clostridia. The acetate-consuming organisms affiliating with Clostridia all contained the FTFHS gene for formyltetrahydrofolate synthetase, a key enzyme for reductive acetogenesis, indicating that these organisms are possible syntrophic acetate-oxidizing (SAO) bacteria that can facilitate acetate consumption via SAO, coupled with hydrogenotrophic methanogenesis (SAO-HM). This study represents the first study applying protein-SIP for analysis of complex biogas samples, a promising method for identifying key microorganisms utilizing specific pathways.

  9. Identification of syntrophic acetate-oxidizing bacteria in anaerobic digesters by combined protein-based stable isotope probing and metagenomics

    Science.gov (United States)

    Mosbæk, Freya; Kjeldal, Henrik; Mulat, Daniel G; Albertsen, Mads; Ward, Alastair J; Feilberg, Anders; Nielsen, Jeppe L

    2016-01-01

    Inhibition of anaerobic digestion through accumulation of volatile fatty acids occasionally occurs as the result of unbalanced growth between acidogenic bacteria and methanogens. A fast recovery is a prerequisite for establishing an economical production of biogas. However, very little is known about the microorganisms facilitating this recovery. In this study, we investigated the organisms involved by a novel approach of mapping protein-stable isotope probing (protein-SIP) onto a binned metagenome. Under simulation of acetate accumulation conditions, formations of 13C-labeled CO2 and CH4 were detected immediately following incubation with [U-13C]acetate, indicating high turnover rate of acetate. The identified 13C-labeled peptides were mapped onto a binned metagenome for improved identification of the organisms involved. The results revealed that Methanosarcina and Methanoculleus were actively involved in acetate turnover, as were five subspecies of Clostridia. The acetate-consuming organisms affiliating with Clostridia all contained the FTFHS gene for formyltetrahydrofolate synthetase, a key enzyme for reductive acetogenesis, indicating that these organisms are possible syntrophic acetate-oxidizing (SAO) bacteria that can facilitate acetate consumption via SAO, coupled with hydrogenotrophic methanogenesis (SAO-HM). This study represents the first study applying protein-SIP for analysis of complex biogas samples, a promising method for identifying key microorganisms utilizing specific pathways. PMID:27128991

  10. Replacing soybean meal with gelatin extracted from cow skin and corn protein concentrate as a protein source in broiler diets.

    Science.gov (United States)

    Khalaji, S; Manafi, M; Olfati, Z; Hedyati, M; Latifi, M; Veysi, A

    2016-02-01

    Two experiments were conducted to investigate the effects of replacing soybean meal with gelatin extracted from cow skin and corn protein concentrate as a protein source in broiler diets. Experiments were carried out as a completely randomized design where each experiment involved 4 treatments of 6 replicates and 10 chicks in each pen. Soybean meal proteins in a corn-soy control diet were replaced with 15, 30, and 45% of cow skin gelatin (CSG) or corn protein concentrate (CPC), respectively, in experiments 1 and 2. BW and cumulative feed intake were measured at 7, 21, and 42 d of age. Blood characteristics, relative organs weight and length, ileal digesta viscosity, ileal morphology, and cecal coliform and Salmonella population were measured at 42 d of age. Apparent total tract digestibility of protein was determined during 35 to 42 d of age. Replacement of soybean meal with CSG severely inhibited BW gain, decreased feed intake, and increased FCR in broilers during the experimental period (P ≤ 0.01). The inclusion of CPC reduced BW and increased FCR significantly (P ≤ 0.05) at 21 and 42 d of age without any consequence in feed intake. Protein digestibility was reduced and ileal digesta viscosity was increased linearly by increasing the amount of CSG and CPC in the control diet (P ≤ 0.01). Replacement of soybean meal with CSG and CPC did not significantly alter blood cell profile and plasma phosphorus, creatinine, blood urea nitrogen, Aspartate transaminase, and HDL and LDL cholesterol concentration. The inclusion of CSG linearly (P ≤ 0.05) increased plasma uric acid concentration and alkaline phosphatase activity. Triglyceride and cholesterol levels were decreased significantly (P ≤ 0.05) when the amount of CSG replacement was 15%. The results of this experiment showed that using CSG and CPC negatively affects broiler performance and therefore is not a suitable alternative to soybean meal in commercial diets. © 2015 Poultry Science Association Inc.

  11. Brown pigment formation in heated sugar-protein mixed suspensions containing unmodified and peptically modified whey protein concentrates.

    Science.gov (United States)

    Rongsirikul, Narumol; Hongsprabhas, Parichat

    2016-01-01

    Commercial whey protein concentrate (WPC) was modified by heating the acidified protein suspensions (pH 2.0) at 80 °C for 30 min and treating with pepsin at 37 °C for 60 min. Prior to spray-drying, such modification did not change the molecular weights (MWs) of whey proteins determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). After spray-drying the modified whey protein concentrate with trehalose excipient (MWPC-TH), it was found that the α-lactalbumin (α-La) was the major protein that was further hydrolyzed the most. The reconstituted MWPC-TH contained β-lactoglobulin (β-Lg) as the major protein and small molecular weight (MW) peptides of less than 6.5 kDa. The reconstituted MWPC-TH had higher NH2 group, Trolox equivalent antioxidant capacity (TEAC), lower exposed aromatic ring and thiol (SH) contents than did the commercial WPC. Kinetic studies revealed that the addition of MWPC-TH in fructose-glycine solution was able to reduce brown pigment formation in the mixtures heated at 80 to 95 °C by increasing the activation energy (Ea) of brown pigment formation due to the retardation of fluoresced advanced glycation end product (AGEs) formation. The addition of MWPC to reducing sugar-glycine/commercial WPC was also able to lower brown pigment formation in the sterilized (121 °C, 15 min) mixed suspensions containing 0.1 M reducing sugar and 0.5-1.0 % glycine and/or commercial (P < 0.05). It was demonstrated that the modification investigated in this study selectively hydrolyzed α-La and retained β-Lg for the production of antibrowning whey protein concentrate.

  12. Replacement of fish meal by protein soybean concentrate in practical diets for Pacific white shrimp

    Directory of Open Access Journals (Sweden)

    Mariana Soares

    2015-10-01

    Full Text Available ABSTRACTThe objective of this work was to evaluate the performance of Litopenaeus vannameifed different levels (0, 25, 50, 75, and 100% of soybean protein concentrate (63.07% crude protein, CP to replace fish meal-by product (61.24% CP. The study was conducted in clear water in fifteen 800 L tanks equipped with aeration systems, constant heating (29 ºC, and daily water exchange (30%. Each tank was stocked with 37.5 shrimp/m3 (3.03±0.14 g. Feed was supplied four times a day, at 6% of the initial biomass, adjusted daily. After 42 days, the weight gain of shrimp fed diets with 0 and 25% protein replacement was higher than that observed in shrimp fed 100% replacement, and there were no differences among those fed the other diets. Feed efficiency and survival did not differ among shrimp fed different protein replacements. There was a negative linear trend for growth parameters and feed intake as protein replacement with soybean protein concentrate increased. Fish meal by-product can be replaced by up to 75% of soybean protein concentrate, with no harm to the growth of Pacific white shrimp.

  13. DC biased low-frequency insulating constriction dielectrophoresis for protein biomolecules concentration.

    Science.gov (United States)

    Zhang, Peng; Liu, Yuxin

    2017-09-01

    Sample enrichment or molecules concentration is considered an essential step in sample processing of miniaturized devices aimed at biosensing and bioanalysis. Among all the means involved to achieve this aim, dielectrophoresis (DEP) is increasingly employed in molecules manipulation and concentration because it is non-destructive and high efficiency. This paper presents a methodology to achieve protein concentration utilizing the combination effects of electrokinetics and low frequency insulating dielectrophoresis (iDEP) generated within a microfluidic device, in which a submicron constricted channel was fabricated using DNA molecular combing and replica molding. This fabrication technique avoids using e-beam lithography or other complicated nanochannel fabrication methods, and provides an easy and low cost approach with the flexibility of controlling channel dimensions to create highly constricted channels embedded in a microfluidic device. With theoretical analysis and experiments, we demonstrated that fluorescein isothiocyanate conjugated bovine serum albumin (FITC-BSA) protein molecules can be significantly concentrated to form an arc-shaped band near the constricted channel under the effects of a negative dielectrophoretic force and DC electrokinetic forces within a short period of time. It was also observed that the amplitudes of the applied DC and AC electric fields, the AC frequencies as well as the suspending medium conductivities had strong effects on the concentration responses of the FITC-BSA molecules, including the concentrated area and position, intensities of the focused molecules, and concentration speed. Our method provides a simple and flexible approach for quickly concentrating protein molecules by controlling the applied electric field parameters. The iDEP device reported in this paper can be used as a stand-alone sensor or worked as a pre-concentration module integrated with biosensors for protein biomarker detection. Furthermore, low

  14. Paper-based microfluidic approach for surface-enhanced raman spectroscopy and highly reproducible detection of proteins beyond picomolar concentration.

    Science.gov (United States)

    Saha, Arindam; Jana, Nikhil R

    2015-01-14

    Although microfluidic approach is widely used in various point of care diagnostics, its implementation in surface enhanced Raman spectroscopy (SERS)-based detection is challenging. This is because SERS signal depends on plasmonic nanoparticle aggregation induced generation of stable electromagnetic hot spots and in currently available microfluidic platform this condition is difficult to adapt. Here we show that SERS can be adapted using simple paper based microfluidic system where both the plasmonic nanomaterials and analyte are used in mobile phase. This approach allows analyte induced controlled particle aggregation and electromagnetic hot spot generation inside the microfluidic channel with the resultant SERS signal, which is highly reproducible and sensitive. This approach has been used for reproducible detection of protein in the pico to femtomolar concentration. Presented approach is simple, rapid, and cost-effective, and requires low sample volume. Method can be extended for SERS-based detection of other biomolecules.

  15. Effect of protein binding on unbound atazanavir and darunavir cerebrospinal fluid concentrations.

    Science.gov (United States)

    Delille, Cecile A; Pruett, Sarah T; Marconi, Vincent C; Lennox, Jeffrey L; Armstrong, Wendy S; Arrendale, Richard F; Sheth, Anandi N; Easley, Kirk A; Acosta, Edward P; Vunnava, Aswani; Ofotokun, Ighovwerha

    2014-09-01

    HIV-1 protease inhibitors (PIs) exhibit different protein binding affinities and achieve variable plasma and tissue concentrations. Degree of plasma protein binding may impact central nervous system penetration. This cross-sectional study assessed cerebrospinal fluid (CSF) unbound PI concentrations, HIV-1 RNA, and neopterin levels in subjects receiving either ritonavir-boosted darunavir (DRV), 95% plasma protein bound, or atazanavir (ATV), 86% bound. Unbound PI trough concentrations were measured using rapid equilibrium dialysis and liquid chromatography/tandem mass spectrometry. Plasma and CSF HIV-1 RNA and neopterin were measured by Ampliprep/COBAS® Taqman® 2.0 assay (Roche) and enzyme-linked immunosorbent assay (ALPCO), respectively. CSF/plasma unbound drug concentration ratio was higher for ATV, 0.09 [95% confidence interval (CI) 0.06-0.12] than DRV, 0.04 (95%CI 0.03-0.06). Unbound CSF concentrations were lower than protein adjusted wild-type inhibitory concentration-50 (IC50 ) in all ATV and 1 DRV-treated subjects (P < 0.001). CSF HIV-1 RNA was detected in 2/15 ATV and 4/15 DRV subjects (P = 0.65). CSF neopterin levels were low and similar between arms. ATV relative to DRV had higher CSF/plasma unbound drug ratio. Low CSF HIV-1 RNA and neopterin suggest that both regimens resulted in CSF virologic suppression and controlled inflammation. © 2014, The American College of Clinical Pharmacology.

  16. Ground level environmental protein concentrations in various ecuadorian environments: potential uses of aerosolized protein for ecological research

    Science.gov (United States)

    Staton, Sarah J.R.; Woodward, Andrea; Castillo, Josemar A.; Swing, Kelly; Hayes, Mark A.

    2014-01-01

    Large quantities of free protein in the environment and other bioaerosols are ubiquitous throughout terrestrial ground level environments and may be integrative indicators of ecosystem status. Samples of ground level bioaerosols were collected from various ecosystems throughout Ecuador, including pristine humid tropical forest (pristine), highly altered secondary humid tropical forest (highly altered), secondary transitional very humid forest (regrowth transitional), and suburban dry montane deforested (suburban deforested). The results explored the sensitivity of localized aerosol protein concentrations to spatial and temporal variations within ecosystems, and their value for assessing environmental change. Ecosystem specific variations in environmental protein concentrations were observed: pristine 0.32 ± 0.09 μg/m3, highly altered 0.07 ± 0.05 μg/m3, regrowth transitional 0.17 ± 0.06 μg/m3, and suburban deforested 0.09 ± 0.04 μg/m3. Additionally, comparisons of intra-environmental differences in seasonal/daily weather (dry season 0.08 ± 0.03 μg/m3 and wet season 0.10 ± 0.04 μg/m3), environmental fragmentation (buffered 0.19 ± 0.06 μg/m3 and edge 0.15 ± 0.06 μg/m3), and sampling height (ground level 0.32 ± 0.09 μg/m3 and 10 m 0.24 ± 0.04 μg/m3) demonstrated the sensitivity of protein concentrations to environmental conditions. Local protein concentrations in altered environments correlated well with satellite-based spectral indices describing vegetation productivity: normalized difference vegetation index (NDVI) (r2 = 0.801), net primary production (NPP) (r2 = 0.827), leaf area index (LAI) (r2 = 0.410). Moreover, protein concentrations distinguished the pristine site, which was not differentiated in spectral indices, potentially due to spectral saturation typical of highly vegetated environments. Bioaerosol concentrations represent an inexpensive method to increase understanding of environmental changes, especially in densely vegetated

  17. Molecular Effects of Concentrated Solutes on Protein Hydration, Dynamics, and Electrostatics.

    Science.gov (United States)

    Abriata, Luciano A; Spiga, Enrico; Peraro, Matteo Dal

    2016-08-23

    Most studies of protein structure and function are performed in dilute conditions, but proteins typically experience high solute concentrations in their physiological scenarios and biotechnological applications. High solute concentrations have well-known effects on coarse protein traits like stability, diffusion, and shape, but likely also perturb other traits through finer effects pertinent at the residue and atomic levels. Here, NMR and molecular dynamics investigations on ubiquitin disclose variable interactions with concentrated solutes that lead to localized perturbations of the protein's surface, hydration, electrostatics, and dynamics, all dependent on solute size and chemical properties. Most strikingly, small polar uncharged molecules are sticky on the protein surface, whereas charged small molecules are not, but the latter still perturb the internal protein electrostatics as they diffuse nearby. Meanwhile, interactions with macromolecular crowders are favored mainly through hydrophobic, but not through polar, surface patches. All the tested small solutes strongly slow down water exchange at the protein surface, whereas macromolecular crowders do not exert such strong perturbation. Finally, molecular dynamics simulations predict that unspecific interactions slow down microsecond- to millisecond-timescale protein dynamics despite having only mild effects on pico- to nanosecond fluctuations as corroborated by NMR. We discuss our results in the light of recent advances in understanding proteins inside living cells, focusing on the physical chemistry of quinary structure and cellular organization, and we reinforce the idea that proteins should be studied in native-like media to achieve a faithful description of their function. Copyright © 2016 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  18. Functional properties and sensory testing of whey protein concentrate sweetened with rebaudioside A

    Directory of Open Access Journals (Sweden)

    Paula Gimenez MILANI

    2016-02-01

    Full Text Available ABSTRACT Objective: To develop a natural dietary product with functional benefits for diabetic patients. Whey protein concentrate was obtained through the separation membrane processes and sweetened with rebaudioside A. This product was submitted to sensory testing in humans and used to evaluate possible functional properties in male Wistar rats models with diabetesMellitus induced by streptozotocin. Methods: Two concentrates were produced. Only the second showed protein content of 74.3 and 17.3% of lactose was used as supplementation in induced diabetic rats. This concentrate was obtained from the concentration by reverse osmosis system (180 k Daltons, followed by nanofiltration in a 500 k Daltons membrane and spray drying at 5.0% solution of the first concentrate developed. The concentrate was sweetened with rebaudioside A (rebaudioside A 26 mg/100 g concentrate. All procedures were performed at the Center for Studies in Natural Products, at the Universidade Estadual de Maringá. Three experimental groups were established (n=6: two groups of diabetic animals, one control group and one supplemented group; and a control group of normal mice (non-diabetic. The supplemented group received concentrates sweetened with rebaudioside A in a dose of 100 mg/kg bw/day by an esophageal tube for 35 days. Fasting, the fed state and body weight were assessed weekly for all groups. At the end of the supplementation period, the following were analyzed: plasma parameters of glucose, total cholesterol, triglycerides and fructosamine; the serum levels of aspartate aminotransferase and alanine aminotransferase, water and food intake. Organs and tissues were removed and weighed to assess mass and anatomical changes. Results: The product presented 74% of proteins and 17% of lactose and showed satisfactory sensory testing by the addition of 26 mg of rebaudioside A/100 g concentrate. Supplementation of the product reduced hyperglycemia, plasma fructosamine levels

  19. Determination of adsorbed protein concentration in aluminum hydroxide suspensions by near-infrared transmittance Spectroscopy

    DEFF Research Database (Denmark)

    Lai, Xuxin; Zheng, Yiwu; Jacobsen, Susanne

    2008-01-01

    , using the partial least square regression (PLSR) method to construct a calibration model. The linear concentration range of adsorbed BSA is from 0 to 1.75 mg/mL by using 10 mm path length cuvettes. The influence of the sedimentation in suspension, different buffers, and different aluminum hydroxide......Analysis of aluminum hydroxide based vaccines is difficult after antigen adsorption. Adsorbed protein is often assessed by measuring residual unadsorbed protein for quality control. A new method for the direct determination of adsorbed protein concentration in suspension using near-infrared (NIR......) transmittance spectroscopy is proposed here. A simple adsorption system using albumin from bovine serum (BSA) and aluminum hydroxide as a model system is employed. The results show that the NIR absorbance at 700-1300 nm is correlated to the adsorbed BSA concentration, measured by the ultraviolet (UV) method...

  20. Diagnostic Accuracy of Urine Protein/Creatinine Ratio Is Influenced by Urine Concentration

    Science.gov (United States)

    Yang, Chih-Yu; Chen, Fu-An; Chen, Chun-Fan; Liu, Wen-Sheng; Shih, Chia-Jen; Ou, Shuo-Ming; Yang, Wu-Chang; Lin, Chih-Ching; Yang, An-Hang

    2015-01-01

    Background The usage of urine protein/creatinine ratio to estimate daily urine protein excretion is prevalent, but relatively little attention has been paid to the influence of urine concentration and its impact on test accuracy. We took advantage of 24-hour urine collection to examine both urine protein/creatinine ratio (UPCR) and daily urine protein excretion, with the latter as the reference standard. Specific gravity from a concomitant urinalysis of the same urine sample was used to indicate the urine concentration. Methods During 2010 to 2014, there were 540 adequately collected 24h urine samples with protein concentration, creatinine concentration, total volume, and a concomitant urinalysis of the same sample. Variables associated with an accurate UPCR estimation were determined by multivariate linear regression analysis. Receiver operating characteristic (ROC) curves were generated to determine the discriminant cut-off values of urine creatinine concentration for predicting an accurate UPCR estimation in either dilute or concentrated urine samples. Results Our findings indicated that for dilute urine, as indicated by a low urine specific gravity, UPCR is more likely to overestimate the actual daily urine protein excretion. On the contrary, UPCR of concentrated urine is more likely to result in an underestimation. By ROC curve analysis, the best cut-off value of urine creatinine concentration for predicting overestimation by UPCR of dilute urine (specific gravity ≦ 1.005) was ≦ 38.8 mg/dL, whereas the best cut-off values of urine creatinine for predicting underestimation by UPCR of thick urine were ≧ 63.6 mg/dL (specific gravity ≧ 1.015), ≧ 62.1 mg/dL (specific gravity ≧ 1.020), ≧ 61.5 mg/dL (specific gravity ≧ 1.025), respectively. We also compared distribution patterns of urine creatinine concentration of 24h urine cohort with a concurrent spot urine cohort and found that the underestimation might be more profound in single voided samples

  1. Diagnostic Accuracy of Urine Protein/Creatinine Ratio Is Influenced by Urine Concentration.

    Science.gov (United States)

    Yang, Chih-Yu; Chen, Fu-An; Chen, Chun-Fan; Liu, Wen-Sheng; Shih, Chia-Jen; Ou, Shuo-Ming; Yang, Wu-Chang; Lin, Chih-Ching; Yang, An-Hang

    2015-01-01

    The usage of urine protein/creatinine ratio to estimate daily urine protein excretion is prevalent, but relatively little attention has been paid to the influence of urine concentration and its impact on test accuracy. We took advantage of 24-hour urine collection to examine both urine protein/creatinine ratio (UPCR) and daily urine protein excretion, with the latter as the reference standard. Specific gravity from a concomitant urinalysis of the same urine sample was used to indicate the urine concentration. During 2010 to 2014, there were 540 adequately collected 24h urine samples with protein concentration, creatinine concentration, total volume, and a concomitant urinalysis of the same sample. Variables associated with an accurate UPCR estimation were determined by multivariate linear regression analysis. Receiver operating characteristic (ROC) curves were generated to determine the discriminant cut-off values of urine creatinine concentration for predicting an accurate UPCR estimation in either dilute or concentrated urine samples. Our findings indicated that for dilute urine, as indicated by a low urine specific gravity, UPCR is more likely to overestimate the actual daily urine protein excretion. On the contrary, UPCR of concentrated urine is more likely to result in an underestimation. By ROC curve analysis, the best cut-off value of urine creatinine concentration for predicting overestimation by UPCR of dilute urine (specific gravity ≦ 1.005) was ≦ 38.8 mg/dL, whereas the best cut-off values of urine creatinine for predicting underestimation by UPCR of thick urine were ≧ 63.6 mg/dL (specific gravity ≧ 1.015), ≧ 62.1 mg/dL (specific gravity ≧ 1.020), ≧ 61.5 mg/dL (specific gravity ≧ 1.025), respectively. We also compared distribution patterns of urine creatinine concentration of 24h urine cohort with a concurrent spot urine cohort and found that the underestimation might be more profound in single voided samples. The UPCR in samples with low

  2. Caffeic Acid-PLGA Conjugate to Design Protein Drug Delivery Systems Stable to Irradiation

    Directory of Open Access Journals (Sweden)

    Francesca Selmin

    2015-01-01

    Full Text Available This work reports the feasibility of caffeic acid grafted PLGA (g-CA-PLGA to design biodegradable sterile microspheres for the delivery of proteins. Ovalbumin (OVA was selected as model compound because of its sensitiveness of γ-radiation. The adopted grafting procedure allowed us to obtain a material with good free radical scavenging properties, without a significant modification of Mw and Tg of the starting PLGA (Mw PLGA = 26.3 ± 1.3 kDa vs. Mw g-CA-PLGA = 22.8 ± 0.7 kDa; Tg PLGA = 47.7 ± 0.8 °C vs. Tg g-CA-PLGA = 47.4 ± 0.2 °C. By using a W1/O/W2 technique, g-CA-PLGA improved the encapsulation efficiency (EE, suggesting that the presence of caffeic residues improved the compatibility between components (EEPLGA = 35.0% ± 0.7% vs. EEg-CA-PLGA = 95.6% ± 2.7%. Microspheres particle size distribution ranged from 15 to 50 µm. The zeta-potential values of placebo and loaded microspheres were −25 mV and −15 mV, respectively. The irradiation of g-CA-PLGA at the dose of 25 kGy caused a less than 1% variation of Mw and the degradation patterns of the non-irradiated and irradiated microspheres were superimposable. The OVA content in g-CA-PLGA microspheres decreased to a lower extent with respect to PLGA microspheres. These results suggest that g-CA-PLGA is a promising biodegradable material to microencapsulate biological drugs.

  3. Microvolume protein concentration determination using the NanoDrop 2000c spectrophotometer.

    Science.gov (United States)

    Desjardins, Philippe; Hansen, Joel B; Allen, Michael

    2009-11-04

    Traditional spectrophotometry requires placing samples into cuvettes or capillaries. This is often impractical due to the limited sample volumes often used for protein analysis. The Thermo Scientific NanoDrop 2000c Spectrophotometer solves this issue with an innovative sample retention system that holds microvolume samples between two measurement surfaces using the surface tension properties of liquids, enabling the quantification of samples in volumes as low as 0.5-2 microL. The elimination of cuvettes or capillaries allows real time changes in path length, which reduces the measurement time while greatly increasing the dynamic range of protein concentrations that can be measured. The need for dilutions is also eliminated, and preparations for sample quantification are relatively easy as the measurement surfaces can be simply wiped with laboratory wipe. This video article presents modifications to traditional protein concentration determination methods for quantification of microvolume amounts of protein using A280 absorbance readings or the BCA colorimetric assay.

  4. Effects of plant proteins on postprandial, free plasma amino acid concentrations in rainbow trout (Oncorhynchus mykiss)

    DEFF Research Database (Denmark)

    Larsen, Bodil Katrine; Dalsgaard, Anne Johanne Tang; Pedersen, Per Bovbjerg

    2012-01-01

    proteins from wheat, peas, field beans, sunflower and soybean. Blood samples were obtained from the caudal vein of 7 fish in each dietary treatment group prior to feeding, as well as: 2, 4, 6, 8, 12, 24, 48 and 72 h after feeding (sampling 7 new fish at each time point), and plasma amino acid......Postprandial patterns in plasma free amino acid concentrations were investigated in juvenile rainbow trout (Oncorhynchus mykiss) fed either a fish meal based diet (FM) or a diet (VEG) where 59% of fish meal protein (corresponding to 46% of total dietary protein) was replaced by a matrix of plant...... the two dietary treatment groups correlated largely with the amino acid content of the two diets except for methionine, lysine and arginine, where the differences were more extreme than what would be expected from differences in dietary concentrations. The apparent protein digestibility coefficient...

  5. Hypercalcemia and high parathyroid hormone-related protein concentration associated with malignant melanoma in a dog.

    Science.gov (United States)

    Pressler, Barrak M; Rotstein, David S; Law, Jerry M; Rosol, Thomas J; LeRoy, Bruce; Keene, Bruce W; Jackson, Mark W

    2002-07-15

    A 12-year-old Cocker Spaniel with an oral malignant melanoma was evaluated for progressive lethargy and anorexia. No metastases were identified during antemortem evaluation, but severe hypercalcemia was evident. Antemortem diagnostic testing failed to identify a cause for the hypercalcemia. No neoplasms other than the melanoma were identified on postmortem examination. Serum parathyroid hormone-related protein concentration was markedly high, and the melanoma had moderate to marked immunostaining for this protein. Paraneoplastic syndromes are rare in dogs with malignant melanoma.

  6. High-temperature stable absorber coatings for linear concentrating solar thermal power plants; Hochtemperaturstabile Absorberschichten fuer linear konzentrierende solarthermische Kraftwerke

    Energy Technology Data Exchange (ETDEWEB)

    Hildebrandt, Christina

    2009-03-23

    This work describes the development of new absorber coatings for different applications - para-bolic trough and linear Fresnel collectors - and operating conditions - absorber in vacuum or in air. The demand for higher efficiencies of solar thermal power plants using parabolic trough technology results in higher temperatures in the collectors and on the absorber tubes. As heat losses increase strongly with increasing temperatures, the need for a lower emissivity of the absorber coating at constant absorptivity arises. The linear Fresnel application envisions ab-sorber tubes stable in air at high temperatures of about 450 C, which are to date commercially not available. This work comprises the theoretical background, the modeling and the fabrication of absorber tubes including the technology transfer to a production-size inline sputter coater. In annealing tests and accompanying optical measurements, degradation processes have been observed and specified more precisely by material characterization techniques. The simulations provided the capability of different materials used as potential IR-reflector. The highest selectivity can be achieved by applying silver which consequently has been chosen for the application in absorber coatings of the parabolic trough technology. Thin silver films how-ever need to be stabilized when used at high temperatures. Appropriate barrier layers as well as process and layer parameters were identified. A high selectivity was achieved and stability of the absorber coating for 1200 h at 500 C in vacuum has been demonstrated. For the application in air, silver was also analyzed as a potential IR-reflector. Even though the stability could be increased considerably, it nevertheless proved to be insufficient. The main factors influencing stability in a positive way are the use of higher quality polishing, additional barrier layers and adequate process parameters. This knowledge was applied for developing coatings which are stable in air at

  7. Enhanced Temperature During Grain Filling Reduces Protein Concentration of Durum Wheat

    Directory of Open Access Journals (Sweden)

    Franco Miglietta

    2011-02-01

    Full Text Available Durum wheat is cultivated over more than 13 millions of hectares (ha world wide and Italy is the main European producer with 3.5 millions tons per year. The protein concentration of durum wheat is very important, it ensures high nutritional value and is highly appreciated by the pasta production industries. The protein concentration of wheat is determined during the grain filling period when carbon and nitrogen compounds are translocated into the grains. Air temperature affects translocation rates and contributes to final protein concentration of wheat grains. Two common commercial varieties of durum and bread wheat were exposed from anthesis to harvest, to a source of infrared radiation in the field. This allowed to investigate the relative effect of temperature on translocation of carbon and nitrogen compound during grain filling. The heat treatment imposed affected marginally dry mass accumulation of the grains in bread wheat and didn’t affect dry mass in durum wheat. Grain protein was affected by heat treatment in durum but not in bread wheat. Carbon accumulation rate was higher for durum than for bread wheat. The protein concentration was greater in durum than in bread wheat and we can assume that the absolute nitrogen accumulation rates were higher for the former species. Such difference may be either caused by a faster nitrogen uptake rate and translocation or a more efficient relocation of nitrogen accumulated in reserve organs.

  8. Turkish Tombul hazelnut (Corylus avellana L.) protein concentrates: functional and rheological properties.

    Science.gov (United States)

    Tatar, F; Tunç, M T; Kahyaoglu, T

    2015-02-01

    Turkish Tombul hazelnut consumed as natural or processed forms were evaluated to obtain protein concentrate. Defatted hazelnut flour protein (DHFP) and defatted hazelnut cake protein (DHCP) were produced from defatted hazelnut flour (DHF) and defatted hazelnut cake (DHC), respectively. The functional properties (protein solubility, emulsifying properties, foaming capacity, and colour), and dynamic rheological characteristics of protein concentrates were measured. The protein contents of samples varied in the range of 35-48 % (w/w, db) and 91-92 % (w/w, db) for DHF/DHC and DHFP/DHCP samples, respectively. The significant difference for water/fat absorption capacity, emulsion stability between DHF and DHC were determined. On the other hand, the solubility and emulsion activity of DHF and DHC were not significantly different (p > 0.05). Emulsion stability of DHFP (%46) was higher than that of DHCP (%35) but other functional properties were found similar. According to these results, the DHCP could be used as DHFP in food product formulations. The DHFP and DHCP samples showed different apparent viscosity at the same temperature and concentration, the elastic modulus (G' value) of DHPC was also found higher than that of DHFP samples.

  9. Dysregulation of autism-associated synaptic proteins by psychoactive pharmaceuticals at environmental concentrations.

    Science.gov (United States)

    Kaushik, Gaurav; Xia, Yu; Pfau, Jean C; Thomas, Michael A

    2017-11-20

    Autism Spectrum Disorders (ASD) are complex neurological disorders for which the prevalence in the U.S. is currently estimated to be 1 in 50 children. A majority of cases of idiopathic autism in children likely result from unknown environmental triggers in genetically susceptible individuals. These triggers may include maternal exposure of a developing embryo to environmentally relevant minute concentrations of psychoactive pharmaceuticals through ineffectively purified drinking water. Previous studies in our lab examined the extent to which gene sets associated with neuronal development were up- and down-regulated (enriched) in the brains of fathead minnows treated with psychoactive pharmaceuticals at environmental concentrations. The aim of this study was to determine whether similar treatments would alter in vitro expression of ASD-associated synaptic proteins on differentiated human neuronal cells. Human SK-N-SH neuroblastoma cells were differentiated for two weeks with 10μM retinoic acid (RA) and treated with environmentally relevant concentrations of fluoxetine, carbamazepine or venlafaxine, and flow cytometry technique was used to analyze expression of ASD-associated synaptic proteins. Data showed that carbamazepine individually, venlafaxine individually and mixture treatment at environmental concentrations significantly altered the expression of key synaptic proteins (NMDAR1, PSD95, SV2A, HTR1B, HTR2C and OXTR). Data indicated that psychoactive pharmaceuticals at extremely low concentrations altered the in vitro expression of key synaptic proteins that may potentially contribute to neurological disorders like ASD by disrupting neuronal development. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Influence of whey protein concentrate addition on textural properties of corn flour extrudates

    Directory of Open Access Journals (Sweden)

    Mladen Brnčić

    2008-05-01

    Full Text Available Texture is an important propertiy of extruded snack products, and depended on extrusion process conditions, raw material properties and various ingredients properties as well. The main purpose of this research was, using twin-screw extrusion, to manufacture a direct expanded extrudate based on mixtures of corn flour and whey protein concentrate with acceptable textural properties. Mixtures were made of corn flour and three different concentrations of whey protein concentrate (7,5 %, 15 %, 22,5 %. Materials were processed in co-rotating twin-screw extruder APV Baker, MPF 50.15 under input conditions: water intake was 10,08 L/h, 12,18 L/h, 14,28 L/h, screw speed was 300 rpm; expansion temperature was 130 °C; feed rate was 70 kg/h. Textural properties: breaking strength index and expansion ratio were determined. Breaking strength index had largest value for the sample with 22,5 % of whey protein concentrate and water intake of 14,28 L/h. Sample with 7,5 % of whey protein concentrate and 10,08 L/h had largest expansion ratio. Calculated textural properties confirmed validity of samples. This results suggest that enrichment of extrudates with wpc addition up to 22,5 % to improve their nutritional value as well as their textural characteristics can be accomplished. Validation of direct expanded extrudates in dependence of its textural properties have shown validity and justification of this research.

  11. Volumetric interpretation of protein adsorption: interfacial packing of protein adsorbed to hydrophobic surfaces from surface-saturating solution concentrations.

    Science.gov (United States)

    Kao, Ping; Parhi, Purnendu; Krishnan, Anandi; Noh, Hyeran; Haider, Waseem; Tadigadapa, Srinivas; Allara, David L; Vogler, Erwin A

    2011-02-01

    The maximum capacity of a hydrophobic adsorbent is interpreted in terms of square or hexagonal (cubic and face-centered-cubic, FCC) interfacial packing models of adsorbed blood proteins in a way that accommodates experimental measurements by the solution-depletion method and quartz-crystal-microbalance (QCM) for the human proteins serum albumin (HSA, 66 kDa), immunoglobulin G (IgG, 160 kDa), fibrinogen (Fib, 341 kDa), and immunoglobulin M (IgM, 1000 kDa). A simple analysis shows that adsorbent capacity is capped by a fixed mass/volume (e.g. mg/mL) surface-region (interphase) concentration and not molar concentration. Nearly analytical agreement between the packing models and experiment suggests that, at surface saturation, above-mentioned proteins assemble within the interphase in a manner that approximates a well-ordered array. HSA saturates a hydrophobic adsorbent with the equivalent of a single square or hexagonally-packed layer of hydrated molecules whereas the larger proteins occupy two-or-more layers, depending on the specific protein under consideration and analytical method used to measure adsorbate mass (solution depletion or QCM). Square or hexagonal (cubic and FCC) packing models cannot be clearly distinguished by comparison to experimental data. QCM measurement of adsorbent capacity is shown to be significantly different than that measured by solution depletion for similar hydrophobic adsorbents. The underlying reason is traced to the fact that QCM measures contribution of both core protein, water of hydration, and interphase water whereas solution depletion measures only the contribution of core protein. It is further shown that thickness of the interphase directly measured by QCM systematically exceeds that inferred from solution-depletion measurements, presumably because the static model used to interpret solution depletion does not accurately capture the complexities of the viscoelastic interfacial environment probed by QCM. Copyright © 2010

  12. Leptospiral outer membrane protein LipL41 is not essential for acute leptospirosis but requires a small chaperone protein, lep, for stable expression.

    Science.gov (United States)

    King, Amy M; Bartpho, Thanatchaporn; Sermswan, Rasana W; Bulach, Dieter M; Eshghi, Azad; Picardeau, Mathieu; Adler, Ben; Murray, Gerald L

    2013-08-01

    Leptospirosis is a worldwide zoonosis caused by pathogenic Leptospira spp., but knowledge of leptospiral pathogenesis remains limited. However, the development of mutagenesis systems has allowed the investigation of putative virulence factors and their involvement in leptospirosis. LipL41 is the third most abundant lipoprotein found in the outer membranes of pathogenic leptospires and has been considered a putative virulence factor. LipL41 is encoded on the large chromosome 28 bp upstream of a small open reading frame encoding a hypothetical protein of unknown function. This gene was named lep, for LipL41 expression partner. In this study, lipL41 was found to be cotranscribed with lep. Two transposon mutants were characterized: a lipL41 mutant and a lep mutant. In the lep mutant, LipL41 protein levels were reduced by approximately 90%. Lep was shown through cross-linking and coexpression experiments to bind to LipL41. Lep is proposed to be a molecular chaperone essential for the stable expression of LipL41. The roles of LipL41 and Lep in the pathogenesis of Leptospira interrogans were investigated; surprisingly, neither of these two unique proteins was essential for acute leptospirosis.

  13. Blood vitamin D(3) metabolite concentrations of adult female bearded dragons (Pogona vitticeps) remain stable after ceasing UVb exposure.

    Science.gov (United States)

    Oonincx, D G A B; van de Wal, M D; Bosch, G; Stumpel, J B G; Heijboer, A C; van Leeuwen, J P T M; Hendriks, W H; Kik, M

    2013-07-01

    Vitamin D deficiency can lead to several health problems collectively called metabolic bone disease (MBD). One commonly kept reptile species prone to develop MBD if managed incorrectly is the bearded dragon (Pogona vitticeps). This study aimed to determine the extent to which adult female bearded dragons fed a diet low in vitamin D can use stored vitamin D and its metabolites to maintain plasma 25(OH)D(3) and 1,25(OH)(2)D(3) concentrations after discontinuing UVb exposure. Blood samples of healthy adult female bearded dragons, exposed to UVb radiation for over 6 months were collected (day 0) after which UVb exposure was discontinued for 83 days and blood was collected. Blood plasma was analysed for concentrations of total Ca, total P, ionized Ca, uric acid, 25(OH)D(3) and 1,25(OH)(2)D(3). There was no significant change in plasma 25(OH)D(3) and 1,25(OH)(2)D(3) concentrations during the study. While total Ca and P in whole blood was found to significantly decrease over time (P dragons, previously exposed to UVb, are able to maintain blood vitamin D metabolite concentrations when UVb exposure is discontinued for a period of up to 83 days. Copyright © 2013 Elsevier Inc. All rights reserved.

  14. Liquid Whey Protein Concentrates Produced by Ultrafiltration as Primary Raw Materials for Thermal Dairy Gels

    Directory of Open Access Journals (Sweden)

    Marta Henriques

    2017-01-01

    Full Text Available The aim of this work is to study the gelation properties of liquid whey protein concentrates (LWPC produced by ultrafiltration (UF as raw material for thermally induced gels intended for food applications. LWPC thermal gelation was performed using different types of LWPC (non-defatted, defatted and diafiltered of different protein mass fractions and pH. Most of the produced gels showed viscoelastic behaviour. Non-defatted LWPC gave stronger heat-induced gels with a more cohesive microstructure, a higher water holding capacity and also higher elastic modulus (G’ and viscous modulus (G’’. Gel properties were not improved in products with lower content of non-protein compounds. As expected, the increase in protein mass fraction positively influences protein interactions. However, the pH is responsible for the equilibrium between attraction and repulsion forces in the gel components that influence gel hardness and water holding capacity.

  15. Passive dosing of triclosan in multi-generation tests with copepods - Stable exposure concentrations and effects at the low µg l-1 range

    DEFF Research Database (Denmark)

    Ribbenstedt, Anton; Mustajärvi, Lukas; Breitholtz, Magnus

    2017-01-01

    to test the applicability of passive dosing to maintain stable concentrations of the organochlorine bacteriocide triclosan in the water phase during a 6-week multi-generation population development test with the harpacticoid copepod Nitocra spinipes. Triclosan was loaded into silicone (1000 mg), which...... was used as passive dosing phase in the exposure vials. The distribution ratio for triclosan between silicone and water (Dsilicone-water ) was 10466 ± 1927. A population development test was conducted at three concentration levels of triclosan that were measured to be 3-5 µg L(-1) , 7-11 µg L(-1) and 16...... exerted on juvenile development. Progressively lower development index values in the populations exposed to increasing triclosan concentrations suggest developmental retardation. Our results further stress the need for chronic exposure during ecotoxicity testing in chemical risk assessment as even...

  16. Effect of ceramic membrane channel diameter on limiting retentate protein concentration during skim milk microfiltration.

    Science.gov (United States)

    Adams, Michael C; Barbano, David M

    2016-01-01

    Our objective was to determine the effect of retentate flow channel diameter (4 or 6mm) of nongraded permeability 100-nm pore size ceramic membranes operated in nonuniform transmembrane pressure mode on the limiting retentate protein concentration (LRPC) while microfiltering (MF) skim milk at a temperature of 50°C, a flux of 55 kg · m(-2) · h(-1), and an average cross-flow velocity of 7 m · s(-1). At the above conditions, the retentate true protein concentration was incrementally increased from 7 to 11.5%. When temperature, flux, and average cross-flow velocity were controlled, ceramic membrane retentate flow channel diameter did not affect the LRPC. This indicates that LRPC is not a function of the Reynolds number. Computational fluid dynamics data, which indicated that both membranes had similar radial velocity profiles within their retentate flow channels, supported this finding. Membranes with 6-mm flow channels can be operated at a lower pressure decrease from membrane inlet to membrane outlet (ΔP) or at a higher cross-flow velocity, depending on which is controlled, than membranes with 4-mm flow channels. This implies that 6-mm membranes could achieve a higher LRPC than 4-mm membranes at the same ΔP due to an increase in cross-flow velocity. In theory, the higher LRPC of the 6-mm membranes could facilitate 95% serum protein removal in 2 MF stages with diafiltration between stages if no serum protein were rejected by the membrane. At the same flux, retentate protein concentration, and average cross-flow velocity, 4-mm membranes require 21% more energy to remove a given amount of permeate than 6-mm membranes, despite the lower surface area of the 6-mm membranes. Equations to predict skim milk MF retentate viscosity as a function of protein concentration and temperature are provided. Retentate viscosity, retentate recirculation pump frequency required to maintain a given cross-flow velocity at a given retentate viscosity, and retentate protein

  17. The concentration of 137Cs and stable Cs in zooplankton in the western North Pacific in relation to their taxonomic composition

    International Nuclear Information System (INIS)

    Kaeriyama, Hideki; Watabe, Teruhisa; Kusakabe, Masashi

    2008-01-01

    The objective of this study is to obtain information on the background level of 137 Cs in zooplankton in the waters close to Rokkasho-mura in Aomori Prefecture, Japan, where a large-scale project has been progressively promoted to establish the nuclear fuel cycle. Prior to the full-scale operation of the spent nuclear fuel reprocessing plant, zooplankton samples were collected in May and October 2005, June 2006, and June 2007 from the surface water (0-5 m depth). The samples were analyzed to determine the concentrations of 137 Cs and stable Cs in relation to taxonomic composition. The numerical abundance of the plankton varied from 253 to 1514 inds. m -3 . Taxonomic composition differed between the two sampling periods. Although copepods formed the most abundant taxonomic group during both seasons, gelatinous plankton (chaetognaths, siphonophores, appendicularians and doliolids) were more abundant in October 2005 than in June 2006 and June 2007. The concentration of 137 Cs in zooplankton varied from 11 to 24 mBq kg-WW -1 . At the same station, the 137 Cs concentration in zooplankton in October 2005 was almost twice as much as that in June 2006, although the concentration of 137 Cs in seawater did not show a difference. The concentration of stable Cs was measured for each taxonomic group: that in gelatinous zooplankton (chaetognaths) was higher than that in crustacean zooplankton (copepods, euphausiids and amphipods). These results suggest that the concentration of 137 Cs in zooplankton is affected by the occurrence of gelatinous zooplankton. (author)

  18. [Activity of alpha-amylase and concentration of protein in saliva of pregnant women].

    Science.gov (United States)

    Ciejak, Magdalena; Olszewska, Maria; Jakubowska, Katarzyna; Zebiełowicz, Dariusz; Safranow, Krzysztof; Chlubek, Dariusz

    2007-01-01

    One of the hypothetical reasons of the increased incidence of caries in women during the pregnancy may be the increased activity of alpha-amylase, which can be found in their saliva. The enzyme takes part in the process of decomposition of simple sugars, which make basic substrate for caries-causing bacteria. The aim of the paper was the evaluation of the influence of pregnancy and gestational age on the activity of alpha-amylase and the concentration of protein in women's saliva. The examined group consisted of 64 pregnant women at age 17-39, between 21st and 40th week of pregnancy. The control group consisted of 44 healthy women at age 20-35, who were not pregnant. In saliva, which was taken before morning meal, without stimulation, protein concentration was determined by Bradford method and the activity of amylase was determined by kinetic method. The activity of amylase correlated strongly and positively with protein concentration in saliva of both the pregnant (RS = +0.65; p women. There were no significant differences between examined parameters in the examined and the control group. It has been observed in the examined group, that there is the significant negative correlation between protein concentration in saliva and the week of pregnancy (RS = -0.35; p increased caries incidence of pregnant women. However, the observed changes of total protein concentration in saliva during pregnancy, suggest that the exact cognition of proteins in pregnant women's saliva may reveal new mechanisms, which lead to an increase of caries risk.

  19. Serum protein concentrations from clinically healthy horses determined by agarose gel electrophoresis.

    Science.gov (United States)

    Riond, Barbara; Wenger-Riggenbach, Bettina; Hofmann-Lehmann, Regina; Lutz, Hans

    2009-03-01

    Serum protein electrophoresis is a useful screening test in equine laboratory medicine. The method can provide valuable information about changes in the concentrations of albumin and alpha-, beta-, and gamma-globulins and thereby help characterize dysproteinemias in equine patients. Reference values for horses using agarose gel as a support medium have not been reported. The purpose of this study was to establish reference intervals for serum protein concentrations in adult horses using agarose gel electrophoresis and to assess differences between warm-blooded and heavy draught horses. In addition, the precision of electrophoresis for determining fraction percentages and the detection limit were determined. Blood samples were obtained from 126 clinically healthy horses, including 105 Thoroughbreds and 21 heavy draught horses of both sexes and ranging from 2 to 20 years of age. The total protein concentration was determined by an automated biuret method. Serum protein electrophoresis was performed using a semi-automated agarose gel electrophoresis system. Coefficients of variation (CVs) were calculated for within-run and within-assay precision. Data from warm-blooded and draught horses were compared using the Mann-Whitney U test. Within-run and within-assay CVs were draught horses and so combined reference intervals (2.5-97.5%) were calculated for total protein (51.0-72.0 g/L), albumin (29.6-38.5 g/L), alpha(1)-globulin (1.9-3.1 g/L), alpha(2)-globulin (5.3-8.7 g/L), beta(1)-globulin (2.8-7.3g/L), beta(2)-globulin (2.2-6.0 g/L), and gamma-globulin (5.8-12.7 g/L) concentrations, and albumin/globulin ratio (0.93-1.65). Using agarose gel as the supporting matrix for serum protein electrophoresis in horses resulted in excellent resolution and accurate results that facilitated standardization into 6 protein fractions.

  20. Gas and particle concentrations in horse stables with individual boxes as a function of the bedding material and the mucking regimen.

    Science.gov (United States)

    Fleming, K; Hessel, E F; Van den Weghe, H F A

    2009-11-01

    The aim of this study was to compare different types of bedding and mucking regimens used in horse stables on the generation of airborne particulate matter bedding material (wheat straw, straw pellets, and wood shavings) used for horses were assessed according to their ammonia generation. Each type of bedding was used for 2 wk, with 3 repetitions. The mean ammonia concentrations within the stable were 3.07 +/- 0.23 mg/m(3) for wheat straw, 4.79 +/- 0.23 mg/m(3) for straw pellets, and 4.27 +/- 0.17 mg/m(3) for wood shavings. In Exp. 2, the effects of the mucking regimen on the generation of ammonia and PM10 from wheat straw (the bedding with the least ammonia generation in the previous experiment) were examined using 3 different daily regimens: 1) no mucking out, 2) complete mucking out, and 3) partial mucking out (removing only feces). The mean ammonia concentrations in the stable differed significantly among all 3 mucking regimens (P bedding regimen without mucking out was evaluated with regard to gas and airborne particle generation. The ammonia values were found not to increase constantly during the course of the 6-wk period. The average weekly values for PM10 also did not increase constantly but varied between approximately 90 and 140 microg/m. It can be concluded from the particle and gas generation patterns found in the results of all 3 experiments that wheat straw was the most suitable bedding of the 3 types investigated and that mucking out completely on a daily basis should not be undertaken in horse stables.

  1. Direct Analysis of Proteins from Solutions with High Salt Concentration Using Laser Electrospray Mass Spectrometry

    Science.gov (United States)

    Karki, Santosh; Shi, Fengjian; Archer, Jieutonne J.; Sistani, Habiballah; Levis, Robert J.

    2018-05-01

    The detection of lysozyme, or a mixture of lysozyme, cytochrome c, and myoglobin, from solutions with varying salt concentrations (0.1 to 250 mM NaCl) is compared using laser electrospray mass spectrometry (LEMS) and electrospray ionization-mass spectrometry (ESI-MS). Protonated protein peaks were observed up to a concentration of 250 mM NaCl in the case of LEMS. In the case of ESI-MS, a protein solution with salt concentration > 0.5 mM resulted in predominantly salt-adducted features, with suppression of the protonated protein ions. The constituents in the mixture of proteins were assignable up to 250 mM NaCl for LEMS and were not assignable above a NaCl concentration of 0.5 mM for ESI. The average sodium adducts () bound to the 7+ charge state of lysozyme for LEMS measurements from salt concentrations of 2.5, 25, 50, and 100 mM NaCl are 1.71, 5.23, 5.26, and 5.11, respectively. The conventional electrospray measurements for lysozyme solution containing salt concentrations of 0.1, 1, 2, and 5 mM NaCl resulted in of 2.65, 6.44, 7.57, and 8.48, respectively. LEMS displays an approximately two orders of magnitude higher salt tolerance in comparison with conventional ESI-MS. The non-equilibrium partitioning of proteins on the surface of the charged droplets is proposed as the mechanism for the high salt tolerance phenomena observed in the LEMS measurements. [Figure not available: see fulltext.

  2. GSTP1, GSTM1 and GSTT1 genetic polymorphisms and total serum GST concentration in stable male COPD

    Directory of Open Access Journals (Sweden)

    Žuntar Irena

    2014-03-01

    Full Text Available The aim of this study was to test the hypothesis that glutathione- S-transferase (GST genotypes were associated with COPD. GSTP1, GSTM1 and GSTT1 genotypes were determined by DNA methods and GST activity spectrophotometrically in older male Caucasian Croats (non- -smokers, ex-smokers, and smokers with stable COPD (n = 30 and sex/age matched controls (n = 60. The distribution of GSTP1 genotypes and alleles in controls vs. COPD showed a statistical difference (p < 0.05. The odds ratio of CC/CT+TT (wild type GSTP1 exon 6 vs. joint heterozygous and mutant homozygous GSTP1 exon 6 was 10.000 and statistically different (p = 0.002. In this study, the GSTP1 mutant genotype of exon 5 (GG, as well as GSTP1 mutant and heterozygous genotypes of exon 6 (TT and CT, were suggested to be genetic contributors to COPD susceptibility. Null GSTM1, null GSTT1 and joint GSTM1/GSTT1 null genotypes were not disease associated. Serum GST was not associated with GST genotypes and COPD or smoking history in our study subjects. Conclusions drawn from the study should be further supported and clarified by studies with larger sample sizes.

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

    Science.gov (United States)

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

    2015-01-01

    To evaluate serum concentrations of biochemical markers and survival time in dogs with protein-losing enteropathy (PLE). Prospective study. 29 dogs with PLE and 18 dogs with food-responsive diarrhea (FRD). Data regarding serum concentrations of various biochemical markers at the initial evaluation were available for 18 of the 29 dogs with PLE and compared with findings for dogs with FRD. Correlations between biochemical marker concentrations and survival time (interval between time of initial evaluation and death or euthanasia) for dogs with PLE were evaluated. Serum C-reactive protein concentration was high in 13 of 18 dogs with PLE and in 2 of 18 dogs with FRD. Serum concentration of canine pancreatic lipase immunoreactivity was high in 3 dogs with PLE but within the reference interval in all dogs with FRD. Serum α1-proteinase inhibitor concentration was less than the lower reference limit in 9 dogs with PLE and 1 dog with FRD. Compared with findings in dogs with FRD, values of those 3 variables in dogs with PLE were significantly different. Serum calprotectin (measured by radioimmunoassay and ELISA) and S100A12 concentrations were high but did not differ significantly between groups. Seventeen of the 29 dogs with PLE were euthanized owing to this disease; median survival time was 67 days (range, 2 to 2,551 days). Serum C-reactive protein, canine pancreatic lipase immunoreactivity, and α1-proteinase inhibitor concentrations differed significantly between dogs with PLE and FRD. Most initial biomarker concentrations were not predictive of survival time in dogs with PLE.

  4. Preparation and physicochemical properties of protein concentrate and isolate produced from Acacia tortilis (Forssk.) Hayne ssp. raddiana.

    Science.gov (United States)

    Embaby, Hassan E; Swailam, Hesham M; Rayan, Ahmed M

    2018-02-01

    The composition and physicochemical properties of defatted acacia flour (DFAF), acacia protein concentrate (APC) and acacia protein isolate (API) were evaluated. The results indicated that API had lower, ash and fat content, than DFAF and APC. Also, significant difference in protein content was noticed among DFAF, APC and API (37.5, 63.7 and 91.8%, respectively). Acacia protein concentrate and isolates were good sources of essential amino acids except cystine and methionine. The physicochemical and functional properties of acacia protein improved with the processing of acacia into protein concentrate and protein isolate. The results of scanning electron micrographs showed that DFAF had a compact structure; protein concentrate were, flaky, and porous type, and protein isolate had intact flakes morphology.

  5. Fermentation of solutions of glucose-protein concentrate in a cascade-multi-ray unit

    Energy Technology Data Exchange (ETDEWEB)

    Denshchikov, M T; Shashilova, V P

    1964-01-01

    Glucose-protein concentrate is a material obtained by the hydrolysis of corn, containing glucose 75 to 80, maltose, isomaltose, and other non-fermentable sugars 1.5 to 2, H/sub 2/O 15 to 17, mineral matter 1.9 to 1%, and N-containing materials 3.2 to 3.4 g/kg. In earlier fermentation trails with this material, after addition of H/sub 2/O, only 10 to 12% ethanol concentrations were obtained. With period addition of citric acid and replacement of the yeast at regular intervals, using a cascade-multitray unit, 12 to 13% concentrations of ethanol were obtained.

  6. Split Nitrogen Application Improves Wheat Baking Quality by Influencing Protein Composition Rather Than Concentration.

    Science.gov (United States)

    Xue, Cheng; Auf'm Erley, Gunda Schulte; Rossmann, Anne; Schuster, Ramona; Koehler, Peter; Mühling, Karl-Hermann

    2016-01-01

    The use of late nitrogen (N) fertilization (N application at late growth stages of wheat, e.g., booting, heading or anthesis) to improve baking quality of wheat has been questioned. Although it increases protein concentration, the beneficial effect on baking quality (bread loaf volume) needs to be clearly understood. Two pot experiments were conducted aiming to evaluate whether late N is effective under controlled conditions and if these effects result from increased N rate or N splitting. Late N fertilizers were applied either as additional N or split from the basal N at late boot stage or heading in the form of nitrate-N or urea. Results showed that late N fertilization improved loaf volume of wheat flour by increasing grain protein concentration and altering its composition. Increasing N rate mainly enhanced grain protein quantitatively. However, N splitting changed grain protein composition by enhancing the percentages of gliadins and glutenins as well as certain high molecular weight glutenin subunits (HMW-GS), which led to an improved baking quality of wheat flour. The late N effects were greater when applied as nitrate-N than urea. The proportions of glutenin and x-type HMW-GS were more important than the overall protein concentration in determining baking quality. N splitting is more effective in improving wheat quality than the increase in the N rate by late N, which offers the potential to cut down N fertilization rates in wheat production systems.

  7. Split nitrogen application improves wheat baking quality by influencing protein composition rather than concentration

    Directory of Open Access Journals (Sweden)

    Cheng eXue

    2016-06-01

    Full Text Available The use of late nitrogen (N fertilization (N application at late growth stages of wheat, e.g. booting, heading or anthesis to improve baking quality of wheat has been questioned. Although it increases protein concentration, the beneficial effect on baking quality (bread loaf volume needs to be clearly understood. Two pot experiments were conducted aiming to evaluate whether late N is effective under controlled conditions and if these effects result from increased N rate or N splitting. Late N fertilizers were applied either as additional N or split from the basal N at late boot stage or heading in the form of nitrate-N or urea. Results showed that late N fertilization improved loaf volume of wheat flour by increasing grain protein concentration and altering its composition. Increasing N rate mainly enhanced grain protein quantitatively. However, N splitting changed grain protein composition by enhancing the percentages of gliadins and glutenins as well as certain high molecular weight glutenin subunits (HMW-GS, which led to an improved baking quality of wheat flour. The late N effects were greater when applied as nitrate-N than urea. The proportions of glutenin and x-type HMW-GS were more important than the overall protein concentration in determining baking quality. N splitting is more effective in improving wheat quality than the increase in the N rate by late N, which offers the potential to cut down N fertilization rates in wheat production systems.

  8. Serum acute phase protein concentrations in female dogs with mammary tumors.

    Science.gov (United States)

    Tecles, Fernando; Caldín, Marco; Zanella, Anna; Membiela, Francisco; Tvarijonaviciute, Asta; Subiela, Silvia Martínez; Cerón, José Joaquín

    2009-03-01

    Acute phase proteins (APPs) are proteins whose concentrations in serum change after any inflammatory stimulus or tissue damage. The aim of the current study was to evaluate 3 positive APPs (C-reactive protein, serum amyloid A, and haptoglobin) and 1 negative APP (albumin) in female dogs with mammary neoplasia. Acute phase proteins were studied in 70 female dogs aged 8-12 years in the following groups: healthy (n = 10); mammary tumors in stages I (n = 19), II (n = 5), III (n = 6), IV (n = 5), and V (n = 7); and with mammary neoplasia plus a concomitant disease (n = 18). In animals with mammary neoplasia, significant increases of positive APPs were only detected in those that had metastasis or a neoplasm with a diameter greater than 5 cm and ulceration. Dogs with mammary neoplasia and a concomitant disease also had high C-reactive protein concentrations. Albumin concentration was decreased in animals with metastasis and with a concomitant disease. The results of the present study indicate that the acute phase response could be stimulated in female dogs with mammary gland tumors because of different factors, such as metastasis, large size of the primary mass, and ulceration or secondary inflammation of the neoplasm.

  9. The elution of certain protein affinity tags with millimolar concentrations of diclofenac.

    Science.gov (United States)

    Baliova, Martina; Juhasova, Anna; Jursky, Frantisek

    2015-12-01

    Diclofenac (2-[(2, 6-dichlorophenyl)amino] benzeneacetic acid) is a sparingly soluble, nonsteroidal anti-inflammatory drug therapeutically acting at low micromolar concentrations. In pH range from 8 to 11, its aqueous solubility can be increased up to 200 times by the presence of counter ions such as sodium. Our protein interaction studies revealed that a millimolar concentration of sodium diclofenac is able to elute glutathione S-transferase (GST), cellulose binding protein (CBD), and maltose binding protein (MBP) but not histidine-tagged or PDZ-tagged proteins from their affinity resins. The elution efficiency of diclofenac is comparable with the eluting agents normally used at similar concentrations. Native gel electrophoresis of sodium diclofenac-treated proteins showed that the interaction is non-covalent and non-denaturing. These results suggest that sodium diclofenac, in addition to its pharmaceutical applications, can also be exploited as a lead for the development of new proteomics reagents. Copyright © 2015 Elsevier B.V. All rights reserved.

  10. Effects of a concentrate of pea antinutritional factors on pea protein digestibility in piglets

    NARCIS (Netherlands)

    Guen, M.P. Le; Huisman, J.; Guéguen, J.; Beelen, G.; Verstegen, M.W.A.

    1995-01-01

    Four experiments were designed to investigate the apparent ileal digestibility of raw pea (Pisum sativum) and two of its components - an isolate of its proteins and a concentrate of its proteinaceous antinutritional factors (ANFs). Three varieties of peas were used: spring varieties Finale and

  11. Quantifying Protein Concentrations Using Smartphone Colorimetry: A New Method for an Established Test

    Science.gov (United States)

    Gee, Clifford T.; Kehoe, Eric; Pomerantz, William C. K.; Penn, R. Lee

    2017-01-01

    Proteins are involved in nearly every biological process, which makes them of interest to a range of scientists. Previous work has shown that hand-held cameras can be used to determine the concentration of colored analytes in solution, and this paper extends the approach to reactions involving a color change in order to quantify protein…

  12. Resting serum concentration of high-sensitivity C-reactive protein ...

    African Journals Online (AJOL)

    Resting serum concentration of high-sensitivity C-reactive protein (hs-CRP) in sportsmen and untrained male adults. F.A. Niyi-Odumosu, O. A. Bello, S.A. Biliaminu, B.V. Owoyele, T.O. Abu, O.L. Dominic ...

  13. Textural behavior of gels formed by rice starch and whey protein isolate: Concentration and crosshead velocities

    Directory of Open Access Journals (Sweden)

    Thiago Novaes Silva

    Full Text Available ABSTRACT Fabricated food gels involving the use of hydrocolloids are gaining polpularity as confectionery/convenience foods. Starch is commonly combined with a hydrocolloid (protein our polyssacharides, particularly in the food industry, since native starches generally do not have ideal properties for the preparation of food products. Therefore the texture studies of starch-protein mixtures could provide a new approach in producing starch-based food products, being thus acritical attribute that needs to be carefully adjusted to the consumer liking. This work investigated the texture and rheological properties of mixed gels of different concentrations of rice starch (15%, 17.5%, and 20% and whey protein isolate (0%, 3%, and 6% with different crosshead velocities (0.05, 5.0, and 10.0 mm/s using a Box-Behnken experimental design. The samples were submitted to uniaxial compression tests with 80% deformation in order to determinate the following rheological parameters: Young’s modulus, fracture stress, fracture deformation, recoverable energy, and apparent biaxial elongational viscosity. Gels with a higher rice starch concentration that were submitted to higher test velocities were more rigid and resistant, while the whey protein isolate concentration had little influence on these properties. The gels showed a higher recoverable energy when the crosshead velocity was higher, and the apparent biaxial elongational viscosity was also influenced by this factor. Therefore, mixed gels exhibit different properties depending on the rice starch concentration and crosshead velocity.

  14. Bioactive Whey Protein Concentrate and Lactose Stimulate Gut Function in Formula-Fed Preterm Pigs

    DEFF Research Database (Denmark)

    Li, Yanqi; Nguyen, Duc Ninh; Ryom, Karina

    2017-01-01

    OBJECTIVE:: Formula feeding is associated with compromised intestinal health in preterm neonates compared with maternal milk, but the mechanisms behind this are unclear. We hypothesized that the use of maltodextrin and whey protein concentrates (WPCs) with reduced bioactivity due to thermal-proce...

  15. Changes in volatile compounds in whey protein concentrate stored at elevated temperature and humidity

    Science.gov (United States)

    Whey protein concentrate (WPC) has been recommended for use in emergency aid programs, but it is often stored overseas without temperature and relative humidity (RH) control, which may cause it to be rejected because of yellowing, off-flavors, or clumping. Therefore, the volatile compounds present ...

  16. The use of crude protein content to predict concentrations of lysine ...

    African Journals Online (AJOL)

    Correlations were determined between the crude protein (CP) and lysine or methionine concentrations of grain from wheat (cultivar: palmiet), barley (cultivar: clipper) and triticale (cultivar: usgen 19) grown in the Western Cape region of South Africa. Twenty samples of varying CP content were collected for each grain type ...

  17. Potential utilization of algal protein concentrate as a food ingredient in space habitats

    Science.gov (United States)

    Nakhost, Z.; Karel, M.

    1989-01-01

    Green alga Scenedesmus obliquus was studied as one of the potential sources of macronutrients in a space habitat. Algal protein concentrate (70.5% protein) was incorporated into a variety of food products such as bran muffins, fettuccine (spinach noodle imitation) and chocolate chip cookies. Food products containing 20 to 40% of incorporated algal proteins were considered. In the sensory analysis the greenish color of the bran muffins and cookies was not found to be objectional. The mild spinachy flavor (algae flavor) was less detectable in chocolate chip cookies than in bran muffins. The color and taste of the algae noodles were found to be pleasant and compared well with commercially available spinach noodles. Commercially available spray-dried Spirulina algae was also incorporated so the products can be compared with those containing Scenedesmus obliquus concentrate. Food products containing commercial algae had a dark green color and a "burnt after taste" and were less acceptable to the panelists.

  18. Simultaneous pre-concentration and separation on simple paper-based analytical device for protein analysis.

    Science.gov (United States)

    Niu, Ji-Cheng; Zhou, Ting; Niu, Li-Li; Xie, Zhen-Sheng; Fang, Fang; Yang, Fu-Quan; Wu, Zhi-Yong

    2018-02-01

    In this work, fast isoelectric focusing (IEF) was successfully implemented on an open paper fluidic channel for simultaneous concentration and separation of proteins from complex matrix. With this simple device, IEF can be finished in 10 min with a resolution of 0.03 pH units and concentration factor of 10, as estimated by color model proteins by smartphone-based colorimetric detection. Fast detection of albumin from human serum and glycated hemoglobin (HBA1c) from blood cell was demonstrated. In addition, off-line identification of the model proteins from the IEF fractions with matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was also shown. This PAD IEF is potentially useful either for point of care test (POCT) or biomarker analysis as a cost-effective sample pretreatment method.

  19. Preservation of grass juice and wet leaf protein concentrate for animal feeds

    Directory of Open Access Journals (Sweden)

    Matti Näsi

    1983-09-01

    Full Text Available Formic acid, mixtures of acids (AIV 1, AIV 2 and formalin-acid mixtures (Viher solution, Viher acid were tested as preservatives of juice and wet leaf protein concentrate (LPC obtained from grass, clover and pea. The main criteria used in judging the success of preservation were changes in the protein fraction, fermentation of sugars, and losses of dry matter and true protein during storage. Fermentation of sugars and moulding could be inhibited in plant juices by adding 0.5 % v/w preservative, but proteolysis continued and true protein was degraded in unheated juices. Ensiling losses of pea juice were considerable, 4.0-15.6 % of DM, in all treatments. For wet leaf protein concentrate precipitated by steaming (85°C, good preservation could be obtained with the additives used in silage making applied at a level of 1 % v/w. In these treatments protein breakdown was minimal, because heating eliminated proteolytic enzymes and partly sterilized the LPC product.

  20. The specific ion effect on emulsions, foam and gels of a seed protein concentrate

    International Nuclear Information System (INIS)

    Lawal, O.S.

    2008-05-01

    Protein concentrate was prepared from the seeds of jack bean (Canavalia ensiformis) and the influences of selected Hofmeister salts on some functional properties of the protein concentrate were investigated. The results indicate that kosmotropic salts (Na 2 SO 4 , NaCl, NaBr) had improved water absorption capacities over the chaotropic salts (NaI, NaClO 4 , NaSCN) and generally, the reduction in water absorption capacity followed the Hofmeister trend: Na 2 SO 4 > NaCl > NaBr > NaI > NaClO 4 > NaSCN. However, the reverse was observed for the foaming and emulsification properties. The least gelation concentration (LGC) was used as the index of gelation properties and the results showed that LGC were higher in kosmotropic salts than in chaotropic salts. Generally, increases in salt concentration reduced the water absorption capacity, the surfactant properties as well as the gelation property. The findings would provide insight into the understanding of the structure property relations of the protein concentrate. (author)

  1. Mercury concentrations in Alaska Pacific halibut muscle relative to stable isotopes of C and N and other biological variables

    International Nuclear Information System (INIS)

    Bentzen, Rebecca; Castellini, J. Margaret; Gerlach, Robert; Dykstra, Claude; O'Hara, Todd

    2016-01-01

    Total mercury concentrations ([THg]), δ 15 N and δ 13 C values were determined in muscle of 693 Pacific halibut caught in International Pacific Halibut Commission setline surveys in Alaska (2002 − 2011). Project goals were to evaluate whether 1) δ 15 N and δ 13 C varied with region, age, sex and length of halibut, and 2) muscle [THg] varied with δ 15 N and δ 13 C (feeding ecology) while accounting for sex, length, and region. Variation in [THg] was explained, in part, by halibut feeding ecology as [THg] increased with trophic position (increasing δ 15 N). Halibut from the western Aleutian Island region were the exception, with overall lower δ 15 N values and significantly higher [THg] than halibut from other Alaskan waters. This [THg] pattern has been observed in other Aleutian biota, possibly the result of northeasterly atmospheric movement of mercury emissions from Asia and/or other local sources and processes. The significantly lower δ 15 N values for these halibut warrants further investigation of halibut prey. - Highlights: • Mercury, δ 15 N, and δ 13 C were determined in muscle of 693 Pacific halibut in Alaska. • Variation in muscle mercury level was explained, in part, by halibut feeding ecology. • Muscle mercury concentration generally increased with relative trophic position (δ 15 N). • Western Aleutian halibut had lower δ 15 N and higher mercury than other regions. • Assessment of mercury and trophic relationships accounted for sex, size, and region.

  2. Meal composition and plasma amino acid ratios: Effect of various proteins or carbohydrates, and of various protein concentrations

    Science.gov (United States)

    Yokogoshi, Hidehiko; Wurtman, Richard J.

    1986-01-01

    The effects of meals containing various proteins and carbohydrates, and of those containing various proportions of protein (0 percent to 20 percent of a meal, by weight) or of carbohydrate (0 percent to 75 percent), on plasma levels of certain large neutral amino acids (LNAA) in rats previously fasted for 19 hours were examined. Also the plasma tryptophan ratios (the ratio of the plasma trytophan concentration to the summed concentrations of the other large neutral amino acids) and other plasma amino acid ratios were calculated. (The plasma tryptophan ratio has been shown to determine brain tryptophan levels and, thereby, to affect the synthesis and release of the neurotransmitter serotonin). A meal containing 70 percent to 75 percent of an insulin-secreting carbohydrate (dextrose or dextrin) increased plasma insulin levels and the tryptophan ratio; those containing 0 percent or 25 percent carbohydrate failed to do so. Addition of as little as 5 percent casein to a 70 percent carbohydrate meal fully blocked the increase in the plasma tryptophan ratio without affecting the secretion of insulin - probably by contributing much larger quantities of the other LNAA than of tryptophan to the blood. Dietary proteins differed in their ability to suppress the carbohydrate-induced rise in the plasma tryptophan ratio. Addition of 10 percent casein, peanut meal, or gelatin fully blocked this increase, but lactalbumin failed to do so, and egg white did so only partially. (Consumption of the 10 percent gelatin meal also produced a major reduction in the plasma tyrosine ratio, and may thereby have affected brain tyrosine levels and catecholamine synthesis.) These observations suggest that serotonin-releasing neurons in brains of fasted rats are capable of distinguishing (by their metabolic effects) between meals poor in protein but rich in carbohydrates that elicit insulin secretion, and all other meals. The changes in brain serotonin caused by carbohydrate-rich, protein

  3. Conservative, unconditionally stable discretization methods for Hamiltonian equations, applied to wave motion in lattice equations modeling protein molecules

    Science.gov (United States)

    LeMesurier, Brenton

    2012-01-01

    A new approach is described for generating exactly energy-momentum conserving time discretizations for a wide class of Hamiltonian systems of DEs with quadratic momenta, including mechanical systems with central forces; it is well-suited in particular to the large systems that arise in both spatial discretizations of nonlinear wave equations and lattice equations such as the Davydov System modeling energetic pulse propagation in protein molecules. The method is unconditionally stable, making it well-suited to equations of broadly “Discrete NLS form”, including many arising in nonlinear optics. Key features of the resulting discretizations are exact conservation of both the Hamiltonian and quadratic conserved quantities related to continuous linear symmetries, preservation of time reversal symmetry, unconditional stability, and respecting the linearity of certain terms. The last feature allows a simple, efficient iterative solution of the resulting nonlinear algebraic systems that retain unconditional stability, avoiding the need for full Newton-type solvers. One distinction from earlier work on conservative discretizations is a new and more straightforward nearly canonical procedure for constructing the discretizations, based on a “discrete gradient calculus with product rule” that mimics the essential properties of partial derivatives. This numerical method is then used to study the Davydov system, revealing that previously conjectured continuum limit approximations by NLS do not hold, but that sech-like pulses related to NLS solitons can nevertheless sometimes arise.

  4. Correlation between C-Reactive Protein in Peripheral Vein and Coronary Sinus in Stable and Unstable Angina

    Energy Technology Data Exchange (ETDEWEB)

    Leite, Weverton Ferreira, E-mail: wfleite@cardiol.br [Instituto do Coração (InCor) do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo (HC-FMUSP), São Paulo, SP (Brazil); Hospital Beneficência Portuguesa de São Paulo, São Paulo, SP (Brazil); Ramires, José Antonio Franchini; Moreira, Luiz Felipe Pinho; Strunz, Célia Maria Cassaro [Instituto do Coração (InCor) do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo (HC-FMUSP), São Paulo, SP (Brazil); Mangione, José Armando [Hospital Beneficência Portuguesa de São Paulo, São Paulo, SP (Brazil)

    2015-03-15

    High sensitivity C-reactive protein (hs-CRP) is commonly used in clinical practice to assess cardiovascular risk. However, a correlation has not yet been established between the absolute levels of peripheral and central hs-CRP. To assess the correlation between serum hs-CRP levels (mg/L) in a peripheral vein in the left forearm (LFPV) with those in the coronary sinus (CS) of patients with coronary artery disease (CAD) and a diagnosis of stable angina (SA) or unstable angina (UA). This observational, descriptive, and cross-sectional study was conducted at the Instituto do Coração, Hospital das Clinicas, Faculdade de Medicina, Universidade de São Paulo, and at the Hospital Beneficência Portuguesa de Sao Paulo, where CAD patients referred to the hospital for coronary angiography were evaluated. Forty patients with CAD (20 with SA and 20 with UA) were included in the study. Blood samples from LFPV and CS were collected before coronary angiography. Furthermore, analysis of the correlation between serum levels of hs-CRP in LFPV versus CS showed a strong linear correlation for both SA (r = 0.993, p < 0.001) and UA (r = 0.976, p < 0.001) and for the entire sample (r = 0.985, p < 0.001). Our data suggest a strong linear correlation between hs-CRP levels in LFPV versus CS in patients with SA and UA.

  5. High performance and thermally stable tandem solar selective absorber coating for concentrated solar thermal power (CSP) application

    Science.gov (United States)

    Prasad, M. Shiva; Kumar, K. K. Phani; Atchuta, S. R.; Sobha, B.; Sakthivel, S.

    2018-05-01

    A novel tandem absorber system (Mn-Cu-Co-Ox-ZrO2/SiO2) developed on an austenitic stainless steel (SS-304) substrate to show an excellent optical performance (αsol: 0.96; ɛ: 0.23@500 °C). In order to achieve this durable tandem, we experimented with two antireflective layers such as ZrO2-SiO2 and nano SiO2 layer on top of Mn-Cu-Co-Ox-ZrO2 layer. We optimized the thickness of antireflective layers to get good tandem system in terms of solar absorptance and emittance. Field emission scanning electron microscopy (FESEM), UV-Vis-NIR and Fourier transform infrared spectroscopy (FTIR) were used to characterize the developed coatings. Finally, the Mn-Cu-Co-Ox-ZrO2/SiO2 exhibits high temperature resistance up to 800 °C, thus allow an increase in the operating temperature of CSP which may lead to high efficiency. We successfully developed a high temperature resistant tandem layer with easy manufacturability at low cost which is an attractive candidate for concentrated solar power generation (CSP).

  6. Effects of ventilation on hyaluronan and protein concentration in pleural liquid of anesthetized and conscious rabbits.

    Science.gov (United States)

    Wang, P M; Lai-Fook, S J

    1998-01-01

    The hypothesis of this study is that pleural lubrication is enhanced by hyaluronan acting as a boundary lubricant in pleural liquid and by pleural filtration as reflected in changes in protein concentration with ventilation. Anesthetized rabbits were injected intravenously with Evans blue dye and ventilated with 100% O2 at either of two levels of ventilation for 6 h. Postmortem values of hyaluronan, total protein, and Evans blue-dyed albumin (EBA) concentrations in pleural liquid were greater at the higher ventilation, consistent with increases in boundary lubrication, pleural membrane permeability, and pleural filtration. To determine whether these effects were caused by hyperoxia or anesthesia, conscious rabbits were ventilated with either 3% CO2 or room air in a box for 6, 12, or 24 h. Similar to the anesthetized rabbits, pleural liquid hyaluronan concentration after 24 h was higher in the conscious rabbits with the hypercapnic-induced greater ventilation. By contrast, the time course of total protein and EBA in pleural liquid was similar in both groups of conscious rabbits, indicating no effect of ventilation on pleural permeability. The increase in pleural liquid hyaluronan concentration might be the result of mesothelial cell stimulation by a ventilation-induced increase in pleural liquid shear stress.

  7. Absence of diurnal variation of C-reactive protein concentrations in healthy human subjects

    Science.gov (United States)

    Meier-Ewert, H. K.; Ridker, P. M.; Rifai, N.; Price, N.; Dinges, D. F.; Mullington, J. M.

    2001-01-01

    BACKGROUND: The concentration of C-reactive protein (CRP) in otherwise healthy subjects has been shown to predict future risk of myocardial infarction and stroke. CRP is synthesized by the liver in response to interleukin-6, the serum concentration of which is subject to diurnal variation. METHODS: To examine the existence of a time-of-day effect for baseline CRP values, we determined CRP concentrations in hourly blood samples drawn from healthy subjects (10 males, 3 females; age range, 21-35 years) during a baseline day in a controlled environment (8 h of nighttime sleep). RESULTS: Overall CRP concentrations were low, with only three subjects having CRP concentrations >2 mg/L. Comparison of raw data showed stability of CRP concentrations throughout the 24 h studied. When compared with cutoff values of CRP quintile derived from population-based studies, misclassification of greater than one quintile did not occur as a result of diurnal variation in any of the subjects studied. Nonparametric ANOVA comparing different time points showed no significant differences for both raw and z-transformed data. Analysis for rhythmic diurnal variation using a method fitting a cosine curve to the group data was negative. CONCLUSIONS: Our data show that baseline CRP concentrations are not subject to time-of-day variation and thus help to explain why CRP concentrations are a better predictor of vascular risk than interleukin-6. Determination of CRP for cardiovascular risk prediction may be performed without concern for diurnal variation.

  8. Concentrated Protein Body Product Derived from Rice Endosperm as an Oral Tolerogen for Allergen-Specific Immunotherapy—A New Mucosal Vaccine Formulation against Japanese Cedar Pollen Allergy

    Science.gov (United States)

    Wakasa, Yuhya; Takagi, Hidenori; Watanabe, Nobumasa; Kitamura, Noriko; Fujiwara, Yoshihiro; Ogo, Yuko; Hayashi, Shimpei; Yang, Lijun; Ohta, Masaru; Thet Tin, Wai Wai; Sekikawa, Kenji; Takano, Makoto; Ozawa, Kenjirou; Hiroi, Takachika; Takaiwa, Fumio

    2015-01-01

    The endoplasmic reticulum-derived type-I protein body (PB-I) from rice endosperm cells is an ideal candidate formulation for the oral delivery of bioencapsulated peptides as tolerogens for allergen-specific immunotherapy. In the present study, PBs containing the deconstructed Japanese cedar pollen allergens Cryptomeria japonica 1 (Cry j 1) and Cry j 2 were concentrated by treatment with thermostable α-amylase at 90°C to remove the starch from milled rice powder, which resulted in a 12.5-fold reduction of dry weight compared to the starting material. The modified Cry j 1 and Cry j 2 antigens in this concentrated PB product were more resistant to enzymatic digestion than those in the milled seed powder despite the absence of intact cell wall and starch, and remained stable for at least 10 months at room temperature without detectable loss or degradation. The high resistance of these allergens could be attributed to changes in protein physicochemical properties induced by the high temperature concentration process, as suggested by the decreased solubility of the antigens and seed proteins in PBs in step-wise-extraction experiments. Confocal microscopy showed that the morphology of antigen-containing PB-Is was preserved in the concentrated PB product. The concentrated PB product induced specific immune tolerance against Cry j 1 and Cry j 2 in mice when orally administered, supporting its potential use as a novel oral tolerogen formulation. PMID:25774686

  9. Concentrated protein body product derived from rice endosperm as an oral tolerogen for allergen-specific immunotherapy--a new mucosal vaccine formulation against Japanese cedar pollen allergy.

    Directory of Open Access Journals (Sweden)

    Yuhya Wakasa

    Full Text Available The endoplasmic reticulum-derived type-I protein body (PB-I from rice endosperm cells is an ideal candidate formulation for the oral delivery of bioencapsulated peptides as tolerogens for allergen-specific immunotherapy. In the present study, PBs containing the deconstructed Japanese cedar pollen allergens Cryptomeria japonica 1 (Cry j 1 and Cry j 2 were concentrated by treatment with thermostable α-amylase at 90°C to remove the starch from milled rice powder, which resulted in a 12.5-fold reduction of dry weight compared to the starting material. The modified Cry j 1 and Cry j 2 antigens in this concentrated PB product were more resistant to enzymatic digestion than those in the milled seed powder despite the absence of intact cell wall and starch, and remained stable for at least 10 months at room temperature without detectable loss or degradation. The high resistance of these allergens could be attributed to changes in protein physicochemical properties induced by the high temperature concentration process, as suggested by the decreased solubility of the antigens and seed proteins in PBs in step-wise-extraction experiments. Confocal microscopy showed that the morphology of antigen-containing PB-Is was preserved in the concentrated PB product. The concentrated PB product induced specific immune tolerance against Cry j 1 and Cry j 2 in mice when orally administered, supporting its potential use as a novel oral tolerogen formulation.

  10. INTERACTION OF NIZKOMETILIROVANNYJ PECTINS WITH A CONCENTRATE OF PROTEINS OF WHEY

    Directory of Open Access Journals (Sweden)

    H. I. Teshaev

    2012-01-01

    Full Text Available Potentiometric titration method was used to study quality complex formation between low methylated pectin and proteins concentrated from whey. It’s shown that at рН>IEP of the lactoglobulin the interaction occurs between negatively charged chains of LM-pectin and positively charged patches of polypeptide chains. The biopolymers ratio had no significant effect on the initial pH of soluble complex formation (pHc; addition of sodium chloride decreased pHc and pK0 of complexes, which linked to electrostatic nature of complex formation between LM-pectin and whey proteins.

  11. Jatropha curcas Protein Concentrate Stimulates Insulin Signaling, Lipogenesis, Protein Synthesis and the PKCα Pathway in Rat Liver.

    Science.gov (United States)

    León-López, Liliana; Márquez-Mota, Claudia C; Velázquez-Villegas, Laura A; Gálvez-Mariscal, Amanda; Arrieta-Báez, Daniel; Dávila-Ortiz, Gloria; Tovar, Armando R; Torres, Nimbe

    2015-09-01

    Jatropha curcas is an oil seed plant that belongs to the Euphorbiaceae family. Nontoxic genotypes have been reported in Mexico. The purpose of the present work was to evaluate the effect of a Mexican variety of J. curcas protein concentrate (JCP) on weight gain, biochemical parameters, and the expression of genes and proteins involved in insulin signaling, lipogenesis, cholesterol and protein synthesis in rats. The results demonstrated that short-term consumption of JCP increased serum glucose, insulin, triglycerides and cholesterol levels as well as the expression of transcription factors involved in lipogenesis and cholesterol synthesis (SREBP-1 and LXRα). Moreover, there was an increase in insulin signaling mediated by Akt phosphorylation and mTOR. JCP also increased PKCα protein abundance and the activation of downstream signaling pathway targets such as the AP1 and NF-κB transcription factors typically activated by phorbol esters. These results suggested that phorbol esters are present in JCP, and that they could be involved in the activation of PKC which may be responsible for the high insulin secretion and consequently the activation of insulin-dependent pathways. Our data suggest that this Mexican Jatropha variety contains toxic compounds that produce negative metabolic effects which require caution when using in the applications of Jatropha-based products in medicine and nutrition.

  12. Calorie Restricted High Protein Diets Downregulate Lipogenesis and Lower Intrahepatic Triglyceride Concentrations in Male Rats

    Directory of Open Access Journals (Sweden)

    Lee M. Margolis

    2016-09-01

    Full Text Available The purpose of this investigation was to assess the influence of calorie restriction (CR alone, higher-protein/lower-carbohydrate intake alone, and combined CR higher-protein/lower-carbohydrate intake on glucose homeostasis, hepatic de novo lipogenesis (DNL, and intrahepatic triglycerides. Twelve-week old male Sprague Dawley rats consumed ad libitum (AL or CR (40% restriction, adequate (10%, or high (32% protein (PRO milk-based diets for 16 weeks. Metabolic profiles were assessed in serum, and intrahepatic triglyceride concentrations and molecular markers of de novo lipogenesis were determined in liver. Independent of calorie intake, 32% PRO tended to result in lower homeostatic model assessment of insulin resistance (HOMA-IR values compared to 10% PRO, while insulin and homeostatic model assessment of β-cell function (HOMA-β values were lower in CR than AL, regardless of protein intake. Intrahepatic triglyceride concentrations were 27.4 ± 4.5 and 11.7 ± 4.5 µmol·g−1 lower (p < 0.05 in CR and 32% PRO compared to AL and 10% PRO, respectively. Gene expression of fatty acid synthase (FASN, stearoyl-CoA destaurase-1 (SCD1 and pyruvate dehydrogenase kinase, isozyme 4 (PDK4 were 45% ± 1%, 23% ± 1%, and 57% ± 1% lower (p < 0.05, respectively, in CR than AL, regardless of protein intake. Total protein of FASN and SCD were 50% ± 1% and 26% ± 1% lower (p < 0.05 in 32% PRO compared to 10% PRO, independent of calorie intake. Results from this investigation provide evidence that the metabolic health benefits associated with CR—specifically reduction in intrahepatic triglyceride content—may be enhanced by consuming a higher-protein/lower-carbohydrate diet.

  13. Parathyroid hormone related protein concentration in human serum and CSF correlates with age.

    Science.gov (United States)

    Kushnir, Mark M; Peterson, Lisa K; Strathmann, Frederick G

    2018-02-01

    Parathyroid Hormone-Related Protein (PTHrP) is involved in intracellular calcium (Ca) regulation, and has been demonstrated to participate in regulation of Ca in brain cells, activation of neurons, and modulation of pain. However, there are conflicting reports regarding the presence of PTHrP in CSF. PTHrP and Ca were quantified in paired CSF and serum samples using mass spectrometry-based methods. Associations between PTHrP and Ca concentrations with age, sex and concentrations of nine CSF diagnostic markers in a set of 140 paired serum and CSF patient samples were evaluated. The observed median PTHrP concentration in CSF was 51 times higher than in serum; the median concentration of Ca in CSF was 1.8 times lower than in serum. We observed positive correlation between concentrations of PTHrP in CSF and serum (p=0.013). Distribution of PTHrP concentrations in serum was associated with age (p=0.0068) and the concentrations were higher in women. In samples with serum calcium concentrations within the reference intervals (n=118), central 95% distribution of concentrations for Ca-CSF, PTHrP-serum and PTHrP-CSF were 5.4 (4.5-6.1) mg/dL, 1.2 (0.5-2.5) pmol/L, 62 (22-125) pmol/L, respectively. Our data demonstrate that PTHrP is a normal constituent of human CSF with median concentrations 51 fold higher than in serum. Elevated serum PTHrP concentrations were positively correlated with age and significantly higher in women. Our data suggest that CSF could be a significant source of circulating PTHrP. Copyright © 2017 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  14. Effect of prescribing a high protein diet and increasing the dose of dialysis on nutrition in stable chronic haemodialysis patients : a randomized, controlled trial

    NARCIS (Netherlands)

    Kloppenburg, Wybe; Stegeman, CA; Kremer Hovinga, T; Vastenburg, G; Vos, P; de Jong, PE; Huisman, RM

    Background. Protein requirements in stable, adequately dialysed haemodialysis patients are not known and recommendations vary. It is not known whether increasing the dialysis dose above the accepted adequate level has a favourable effect on nutrition. The aim of this study was to determine whether

  15. Dissolved organic carbon, CO2, and CH4 concentrations and their stable isotope ratios in thermokarst lakes on the Qinghai-Tibetan Plateau

    Directory of Open Access Journals (Sweden)

    Cuicui Mu

    2016-01-01

    Full Text Available Thermokarst lakes are widely distributed on the Qinghai-Tibetan Plateau (QTP, which accounts for 8% of the global permafrost area. These lakes probably promote organic matter biodegradation and thus accelerate the emission of carbon-based greenhouse gases. However, little is known about greenhouse gas concentrations and their stable isotopes characteristics of these lakes. In this study, we measured the concentrations of dissolved organic carbon (DOC, dissolved CO2 and CH4, as well as the distribution of δ13CCO2, δ13CCH4, and δ13COM (organic matter of lake sediments in thermokarst lakes on the QTP. Results showed that the OM of the lake sediments was highly decomposed. The concentrations of DOC, CO2 and CH4 in the lake water on the QTP were 1.2–49.6 mg L–1, 3.6–45.0 μmol L–1 and 0.28–3.0 μmol L–1, respectively. The highest CO2 and CH4 concentrations were recorded in July while the lowest values in September, which suggested that temperature had an effect on greenhouse gas production, although this pattern may also relate to thermal stratification of the water column. The results implied that thermokast lakes should be paid more attention to regarding carbon cycle and greenhouse gas emissions on the QTP.

  16. Serum protein concentration in low-dose total body irradiation of normal and malnourished rats

    International Nuclear Information System (INIS)

    Viana, W.C.M.; Lambertz, D.; Borges, E.S.; Neto, A.M.O.; Lambertz, K.M.F.T.; Amaral, A.

    2016-01-01

    Among the radiotherapeutics' modalities, total body irradiation (TBI) is used as treatment for certain hematological, oncological and immunological diseases. The aim of this study was to evaluate the long-term effects of low-dose TBI on plasma concentration of total protein and albumin using prematurely and undernourished rats as animal model. For this, four groups with 9 animals each were formed: Normal nourished (N); Malnourished (M); Irradiated Normal nourished (IN); Irradiated Malnourished (IM). At the age of 28 days, rats of the IN and IM groups underwent total body gamma irradiation with a source of cobalt-60. Total protein and Albumin in the blood serum was quantified by colorimetry. This research indicates that procedures involving low-dose total body irradiation in children have repercussions in the reduction in body-mass as well as in the plasma levels of total protein and albumin. Our findings reinforce the periodic monitoring of total serum protein and albumin levels as an important tool in long-term follow-up of pediatric patients in treatments associated to total body irradiation. - Highlights: • Low-dose total body irradiation (TBI) in children have repercussions in their body-mass. • Long-term total protein and albumin levels are affected by TBI. • The monitoring of total protein and albumin levels are useful in the follow-up of TBI pediatric patients.

  17. Agarose gel electrophoresis of cerebrospinal fluid proteins of dogs after sample concentration using a membrane microconcentrator technique.

    Science.gov (United States)

    Gama, Fernanda Gomes Velasque; Santana, Aureo Evangelista; Filho, Eugênio de Campos; Nogueira, Cláudia Aparecida da Silva

    2007-03-01

    Cerebrospinal fluid (CSF) is produced in the cerebral ventricles through ultrafiltration of plasma and active transport mechanisms. Evaluation of proteins in CSF may provide important information about the production of immunoglobulins within the central nervous system as well as possible disturbances in the blood-brain barrier. The objective of this study was to measure the concentration and fractions of protein in CSF samples using a membrane microconcentrator technique followed by electrophoresis, and to compare the protein fractions obtained with those in serum. CSF samples from 3 healthy dogs and 3 dogs with canine distemper virus infection were concentrated using a membrane microconcentrator having a 0.5 to 30,000 d nominal molecular weight limit (Ultrafree, Millipore, Billerica, MA, USA). Protein concentration was determined before and after concentration. Agarose gel electrophoresis was done on concentrated CSF samples, serum, and serial dilutions of one of the CSF samples. Electrophoretic bands were clearly identified in densitometer tracings in CSF samples with protein concentrations as low as 1.3 g/dL. The higher CSF protein concentration in dogs with distemper was mainly the result of increased albumin concentration. The microconcentrating method used in this study enables characterization of the main protein fractions in CSF by routine electrophoresis and may be useful for interpreting the underlying cause of changes in CSF protein concentrations.

  18. Measurement of the concentration of plasmatic cortisol by competition to the binding protein

    International Nuclear Information System (INIS)

    Okada, H.; Tambascia, M.A.; Wajchenberg, B.L.; Pieroni, R.R.

    1977-01-01

    The concentration of plasmatic cortisol was measured by competition to the binding protein (transcortin), after extracting the samples with dicloromethane. It is a suitable method for clinical routine, 100μl of plasma being used in each analysis. The normal mean +- standard error mean in 8:00 a.m. fasting subjects was 13,62 +- 5,43 μl/100 ml of plasma [pt

  19. Modified procedure for rapid labelling of low concentrations of bioactive proteins with indium-111

    Energy Technology Data Exchange (ETDEWEB)

    Zoghbi, S S; Neumann, R D; Gottschalk, A

    1985-01-01

    The authors describe the conjugation of DTPA to 100-500 g of protein in concentrations of 0.6-1.0 mg mL utilizing the mixed anhydride method. Free DTPA is removed by minicolumn gel filtration and centrifugation with minimal protein dilution. Radiolabelling process can be monitored by instant thin layer chromatography. Any radiochemical impurity detected can be eliminated either by additional minicolumn filtration of further chelation with more conjugated protein. In citrate buffer at pH 6 with minicolumn gel chromatography the authors prepared In-DTPA-D3 (3.0 Ci g) monoclonal antibody and used it to image hepatocarcinoma in guinea pigs. 13 references, 5 figures, 2 tables.

  20. Effect of whey protein concentrate on texture of fat-free desserts: sensory and instrumental measurements

    Directory of Open Access Journals (Sweden)

    Márcia Cristina Teixeira Ribeiro Vidigal

    2012-06-01

    Full Text Available It is important to understand how changes in the product formulation can modify its characteristics. Thus, the objective of this study was to investigate the effect of whey protein concentrate (WPC on the texture of fat-free dairy desserts. The correlation between instrumental and sensory measurements was also investigated. Four formulations were prepared with different WPC concentrations (0, 1.5, 3.0, and 4.5 wt. (% and were evaluated using the texture profile analysis (TPA and rheology. Thickness was evaluated by nine trained panelists. Formulations containing WPC showed higher firmness, elasticity, chewiness, and gumminess and clearly differed from the control as indicated by principal component analysis (PCA. Flow behavior was characterized as time-dependent and pseudoplastic. Formulation with 4.5% WPC at 10 °C showed the highest thixotropic behavior. Experimental data were fitted to Herschel-Bulkley model. The addition of WPC contributed to the texture of the fat-free dairy dessert. The yield stress, apparent viscosity, and perceived thickness in the dairy desserts increased with WPC concentration. The presence of WPC promotes the formation of a stronger gel structure as a result of protein-protein interactions. The correlation between instrumental parameters and thickness provided practical results for food industries.

  1. PerturbationAnalyzer: a tool for investigating the effects of concentration perturbation on protein interaction networks.

    Science.gov (United States)

    Li, Fei; Li, Peng; Xu, Wenjian; Peng, Yuxing; Bo, Xiaochen; Wang, Shengqi

    2010-01-15

    The propagation of perturbations in protein concentration through a protein interaction network (PIN) can shed light on network dynamics and function. In order to facilitate this type of study, PerturbationAnalyzer, which is an open source plugin for Cytoscape, has been developed. PerturbationAnalyzer can be used in manual mode for simulating user-defined perturbations, as well as in batch mode for evaluating network robustness and identifying significant proteins that cause large propagation effects in the PINs when their concentrations are perturbed. Results from PerturbationAnalyzer can be represented in an intuitive and customizable way and can also be exported for further exploration. PerturbationAnalyzer has great potential in mining the design principles of protein networks, and may be a useful tool for identifying drug targets. PerturbationAnalyzer can be accessed from the Cytoscape web site http://www.cytoscape.org/plugins/index.php or http://biotech.bmi.ac.cn/PerturbationAnalyzer. Supplementary data are available at Bioinformatics online.

  2. Chemical Utilization of Albizia lebbeck Leaves for Developing Protein Concentrates as a Dietary Supplement.

    Science.gov (United States)

    Khan, Lutful Haque; Varshney, V K

    2017-08-17

    In search of nonconventional sources of protein to combat widespread malnutrition, the possibility of developing a protein concentrate as an alternative dietary supplement from abundantly available yet poorly valorized leaves of Albizia lebbeck (siris) was examined. A process for recovery of leaf protein concentrate (LPC) from these leaves was optimized and applied for isolation of LPCs from lower, middle, and upper canopies of the tree. The optimized conditions (leaves to water 1:9, coagulation at pH 4.0 using 1 N citric acid at 90°C for 11 minutes) afforded LPCs containing protein 37.15%, 37.57%, and 37.76% in 5.99%, 5.97%, and 6.07% yield, respectively. The proximate nutritional composition, pigments, minerals, in vitro digestibility, and antinutritional factors of these LPCs were determined. Analysis of variance of these data revealed no significant difference with respect to canopy. Use of Albizia lebbeck leaves for development of LPC as a food/feed supplement was revealed.

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

    DEFF Research Database (Denmark)

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

    2004-01-01

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

  4. Type 2 diabetes mellitus is associated with differential effects on plasma cholesteryl ester transfer protein and phospholipid transfer protein activities and concentrations

    NARCIS (Netherlands)

    Dullaart, RPF; De Vries, R; Scheek, L; Borggreve, SE; Van Gent, T; Dallinga-Thie, GM; Ito, M; Nagano, M; Sluiter, WJ; Hattori, H; Van Tol, A

    Background: Human plasma contains two lipid transfer proteins, cholesteryl ester transfer protein (CETP) and phospholipid transfer protein (PLTP), which are crucial in reverse cholesterol transport. Methods: Plasma CETP and PLTP activity levels and concentrations in 16 type 2 diabetic patients and

  5. Adding Sarcosine to Antipsychotic Treatment in Patients with Stable Schizophrenia Changes the Concentrations of Neuronal and Glial Metabolites in the Left Dorsolateral Prefrontal Cortex.

    Science.gov (United States)

    Strzelecki, Dominik; Podgórski, Michał; Kałużyńska, Olga; Stefańczyk, Ludomir; Kotlicka-Antczak, Magdalena; Gmitrowicz, Agnieszka; Grzelak, Piotr

    2015-10-15

    The glutamatergic system is a key point in pathogenesis of schizophrenia. Sarcosine (N-methylglycine) is an exogenous amino acid that acts as a glycine transporter inhibitor. It modulates glutamatergic transmission by increasing glycine concentration around NMDA (N-methyl-d-aspartate) receptors. In patients with schizophrenia, the function of the glutamatergic system in the prefrontal cortex is impaired, which may promote negative and cognitive symptoms. Proton nuclear magnetic resonance (¹H-NMR) spectroscopy is a non-invasive imaging method enabling the evaluation of brain metabolite concentration, which can be applied to assess pharmacologically induced changes. The aim of the study was to evaluate the influence of a six-month course of sarcosine therapy on the concentration of metabolites (NAA, N-acetylaspartate; Glx, complex of glutamate, glutamine and γ-aminobutyric acid (GABA); mI, myo-inositol; Cr, creatine; Cho, choline) in the left dorso-lateral prefrontal cortex (DLPFC) in patients with stable schizophrenia. Fifty patients with schizophrenia, treated with constant antipsychotics doses, in stable clinical condition were randomly assigned to administration of sarcosine (25 patients) or placebo (25 patients) for six months. Metabolite concentrations in DLPFC were assessed with 1.5 Tesla ¹H-NMR spectroscopy. Clinical symptoms were evaluated with the Positive and Negative Syndrome Scale (PANSS). The first spectroscopy revealed no differences in metabolite concentrations between groups. After six months, NAA/Cho, mI/Cr and mI/Cho ratios in the left DLPFC were significantly higher in the sarcosine than the placebo group. In the sarcosine group, NAA/Cr, NAA/Cho, mI/Cr, mI/Cho ratios also significantly increased compared to baseline values. In the placebo group, only the NAA/Cr ratio increased. The addition of sarcosine to antipsychotic therapy for six months increased markers of neurons viability (NAA) and neurogilal activity (mI) with simultaneous improvement

  6. Adding Sarcosine to Antipsychotic Treatment in Patients with Stable Schizophrenia Changes the Concentrations of Neuronal and Glial Metabolites in the Left Dorsolateral Prefrontal Cortex

    Directory of Open Access Journals (Sweden)

    Dominik Strzelecki

    2015-10-01

    Full Text Available The glutamatergic system is a key point in pathogenesis of schizophrenia. Sarcosine (N-methylglycine is an exogenous amino acid that acts as a glycine transporter inhibitor. It modulates glutamatergic transmission by increasing glycine concentration around NMDA (N-methyl-d-aspartate receptors. In patients with schizophrenia, the function of the glutamatergic system in the prefrontal cortex is impaired, which may promote negative and cognitive symptoms. Proton nuclear magnetic resonance (1H-NMR spectroscopy is a non-invasive imaging method enabling the evaluation of brain metabolite concentration, which can be applied to assess pharmacologically induced changes. The aim of the study was to evaluate the influence of a six-month course of sarcosine therapy on the concentration of metabolites (NAA, N-acetylaspartate; Glx, complex of glutamate, glutamine and γ-aminobutyric acid (GABA; mI, myo-inositol; Cr, creatine; Cho, choline in the left dorso-lateral prefrontal cortex (DLPFC in patients with stable schizophrenia. Fifty patients with schizophrenia, treated with constant antipsychotics doses, in stable clinical condition were randomly assigned to administration of sarcosine (25 patients or placebo (25 patients for six months. Metabolite concentrations in DLPFC were assessed with 1.5 Tesla 1H-NMR spectroscopy. Clinical symptoms were evaluated with the Positive and Negative Syndrome Scale (PANSS. The first spectroscopy revealed no differences in metabolite concentrations between groups. After six months, NAA/Cho, mI/Cr and mI/Cho ratios in the left DLPFC were significantly higher in the sarcosine than the placebo group. In the sarcosine group, NAA/Cr, NAA/Cho, mI/Cr, mI/Cho ratios also significantly increased compared to baseline values. In the placebo group, only the NAA/Cr ratio increased. The addition of sarcosine to antipsychotic therapy for six months increased markers of neurons viability (NAA and neurogilal activity (mI with simultaneous

  7. Optimization of expression JTAT protein with emphasis on transformation efficiency and IPTG concentration

    Directory of Open Access Journals (Sweden)

    Endang Tri Margawati

    2017-12-01

    Full Text Available One of small accessory genes between pol and env is tat gene encoding TAT protein. This research was aimed to optimize the expression of Jembrana TAT (JTAT protein with preparing Escherichia coli (E. coli in advance using adopted methods of M1 (MgCl2 + CaCl2 and M2 (CaCl2 + Glycerol. The best transformation efficiency resulting from a better transformation method was used to subsequent expression of JTAT protein. A synthetic tat gene encoding protein JTAT was previously cloned into pBT-hisC. Concentration of 200; 400; 600 µM IPTG was induced to a small volume culture (200 ml; OD600 = 4, incubated for 3 h. Pellets were harvested by centrifugation (4000 rpm; 4 °C; 15 min. Buffer B (10 mM Immidazole was added into pellets, lysed by freeze-thaw followed by sonication. Supernatant was collected by centrifugation (10,000 rpm; 4 °C; 20 min and purified using Ni-NTA Agarose resin, released by elution buffer (E containing 400 mM Immidazole to collect purified protein twice (E1, E2. The protein was characterized by SDS-PAGE and Western Blot (WB, quantified (at λ595 nm with BSA standard method in prior. The result showed that transformation efficiency was better in M2 (2.53 × 106 than M1 (3.10 × 105. The JTAT protein was expressed at a right size of 11.8 kDa. Concentration of 200 µM IPTG produced a significantly better protein yield (1.500 ± 0.089 mg/ml; P < 0.05 than 600 µM IPTG (0.896 ± 0.052 mg/ml and not different to 400 µM IPTG (1.298 ± 0.080 mg/ml. This research indicated that transformation efficiency needs to be taken account in prior of optimization of the protein expression.

  8. Pan-Arctic concentrations of mercury and stable isotope ratios of carbon (δ{sup 13}C) and nitrogen (δ{sup 15}N) in marine zooplankton

    Energy Technology Data Exchange (ETDEWEB)

    Pomerleau, Corinne, E-mail: corinne.pomerleau@umanitoba.ca [Centre for Earth Observation Science, University of Manitoba, Winnipeg, MB R3T 2N2 (Canada); Greenland Institute of Natural Resources, Kivioq 2, Nuuk 3900, Greenland (Denmark); Stern, Gary A.; Pućko, Monika [Centre for Earth Observation Science, University of Manitoba, Winnipeg, MB R3T 2N2 (Canada); Foster, Karen L. [Foster Environmental, Peterborough, ON K9J 8L2 (Canada); Macdonald, Robie W. [Institute of Ocean Sciences, Fisheries and Oceans Canada, Sidney, BC V8L 4B2 (Canada); Fortier, Louis [Québec-Océan, Département de Biologie, Université Laval, Québec, QC G1V 0A6 (Canada)

    2016-05-01

    Zooplankton play a central role in marine food webs, dictating the quantity and quality of energy available to upper trophic levels. They act as “keystone” species in transfer of mercury (Hg) up through the marine food chain. Here, we present the first Pan-Arctic overview of total and monomethylmercury concentrations (THg and MMHg) and stable isotope ratios of carbon (δ{sup 13}C) and nitrogen (δ{sup 15}N) in selected zooplankton species by assembling data collected between 1998 and 2012 from six arctic regions (Laptev Sea, Chukchi Sea, southeastern Beaufort Sea, Canadian Arctic Archipelago, Hudson Bay and northern Baffin Bay). MMHg concentrations in Calanus spp., Themisto spp. and Paraeuchaeta spp. were found to increase with higher δ{sup 15}N and lower δ{sup 13}C. The southern Beaufort Sea exhibited both the highest THg and MMHg concentrations. Biomagnification of MMHg between Calanus spp. and two of its known predators, Themisto spp. and Paraeuchaeta spp., was greatest in the southern Beaufort Sea. Our results show large geographical variations in Hg concentrations and isotopic signatures for individual species related to regional ecosystem features, such as varying water masses and freshwater inputs, and highlight the increased exposure to Hg in the marine food chain of the southern Beaufort Sea. - Highlights: • Assessment of Pan-Arctic variability in zooplankton Hg concentrations • Increased exposure to Hg in the marine food chain of the southern Beaufort Sea • Zooplankton plays a central role in the Hg pathway within Arctic marine food webs.

  9. Interfacial composition and stability of emulsions made with mixtures of commercial sodium caseinate and whey protein concentrate.

    Science.gov (United States)

    Ye, Aiqian

    2008-10-15

    The interfacial composition and the stability of oil-in-water emulsion droplets (30% soya oil, pH 7.0) made with mixtures of sodium caseinate and whey protein concentrate (WPC) (1:1 by protein weight) at various total protein concentrations were examined. The average volume-surface diameter (d32) and the total surface protein concentration of emulsion droplets were similar to those of emulsions made with both sodium caseinate alone and WPC alone. Whey proteins were adsorbed in preference to caseins at low protein concentrations (caseins were adsorbed in preference to whey proteins at high protein concentrations. The creaming stability of the emulsions decreased markedly as the total protein concentration of the system was increased above 2% (sodium caseinate >1%). This was attributed to depletion flocculation caused by the sodium caseinate in these emulsions. Whey proteins did not retard this instability in the emulsions made with mixtures of sodium caseinate and WPC. Copyright © 2008 Elsevier Ltd. All rights reserved.

  10. STUDIES OF METHOD FOR DETERMINING THE PROTEIN CONCENTRATION OF "MALEIN PPD" BY THE KJELDAHL METHOD

    Directory of Open Access Journals (Sweden)

    Ciuca, V

    2017-06-01

    Full Text Available Glanders is a contagious and fatal disease of horses, donkeys, and mules, caused by infection with the bacterium Burkholderia mallei. The pathogen causes nodules and ulcerations in the upper respiratory tract and lungs. Glanders is transmissible to humans by direct contact with diseased animals or with infected or contaminated material. In the untreated acute disease, the mortality rate can reach 95% within 3 weeks Malein PPD - the diagnostic product contain max 2mg/ml Burkholderia mallei. The amount of protein in the biological product "Malein PPD" is measured as nitrogen from protein molecule, applying the Kjeldahl (method determination of nitrogen by sulphuric acid digestion. The validation study aims to demonstrate the determination of the protein of the Malein PPD, by sulphuric acid digestion, it is an appropriate analytical method, reproducible and meets the quality requirements of diagnostic reagents. The paper establishes the performance characteristics of the method considered and identify the factors that influence these characteristics. The method for determining the concentration of protein, by the Kjeldahl method is considered valid if the results obtained for each validation parameter are within the admissibility criteria.The validation procedure includes details on protocol working to determine the protein of the Malein PPD, validation criteria, experimental results, mathematical calculations.

  11. On the interconnection of stable protein complexes: inter-complex hubs and their conservation in Saccharomyces cerevisiae and Homo sapiens networks.

    Science.gov (United States)

    Guerra, Concettina

    2015-01-01

    Protein complexes are key molecular entities that perform a variety of essential cellular functions. The connectivity of proteins within a complex has been widely investigated with both experimental and computational techniques. We developed a computational approach to identify and characterise proteins that play a role in interconnecting complexes. We computed a measure of inter-complex centrality, the crossroad index, based on disjoint paths connecting proteins in distinct complexes and identified inter-complex hubs as proteins with a high value of the crossroad index. We applied the approach to a set of stable complexes in Saccharomyces cerevisiae and in Homo sapiens. Just as done for hubs, we evaluated the topological and biological properties of inter-complex hubs addressing the following questions. Do inter-complex hubs tend to be evolutionary conserved? What is the relation between crossroad index and essentiality? We found a good correlation between inter-complex hubs and both evolutionary conservation and essentiality.

  12. Lipoprotein(a) and dietary proteins: casein lowers lipoprotein(a) concentrations as compared with soy protein1-3

    DEFF Research Database (Denmark)

    Nilausen, Karin Johanne; Meinertz, H.

    1999-01-01

    Lipoprotein(a), plasma lipoproteins, dietary proteins, soy protein, casein, liquid-formula, coronary artery disease, men, Denmark......Lipoprotein(a), plasma lipoproteins, dietary proteins, soy protein, casein, liquid-formula, coronary artery disease, men, Denmark...

  13. Plasma urea nitrogen and progesterone concentrations and follicular dynamics in ewes fed proteins of different degradability

    Directory of Open Access Journals (Sweden)

    Gustavo Bianchi Lazarin

    2012-07-01

    Full Text Available The effects of overfeeding with protein of different degradability on body condition, plasma urea nitrogen and progesterone concentrations, ovulation number and follicular dynamics were assessed in Santa Ines ewes. Twelve ewes were assigned to a randomized block design according to body weight and received overfeeding with soybean meal or with corn gluten meal or maintenance diet for 28 days before ovulation and during the next estrous cycle. Blood samples were taken on days 7, 14, 21, and 28 after the beginning of treatments for analysis of plasma urea nitrogen and on days 3, 6, 9, 12, and 15 into the estrous cycle for analysis of plasma urea nitrogen and progesterone. Follicular dynamics was monitored daily by ultrasound during one estrous cycle. Dry matter and crude protein intake, weight gain, plasma urea nitrogen concentration before ovulation, number of ovulations, diameter of the largest follicle of the 1st and of the 2nd waves and the growth rate of the largest follicle of the 1st wave were higher in the ewes that received overfeeding. The growth rate of the largest follicle of the 3rd wave was higher in the ewes fed maintenance diet. The back fat thickness, plasma urea nitrogen before ovulation and progesterone concentrations, diameter of the largest follicle of the 2nd wave and growth rate of the largest follicle of the 3rd wave were higher in ewes that received overfeeding with soybean meal. The growth rate of the largest follicle of the 1st wave was higher in ewes that received overfeeding with corn gluten meal. Overfeeding with protein-rich feeds may increase the ovulation number and with soybean meal, it may be effective in increasing plasma progesterone concentration in ewes.

  14. Serum C-reactive protein concentration in preeclamptic women: Effect on pregnancy outcome

    Directory of Open Access Journals (Sweden)

    Sharmin Sultana

    2016-07-01

    Full Text Available Background: Preeclampsia is a multisystem disorder of unknown etiology characterized by development of hyperten­sion to the extent of 140/90 mm of Hg or more with proteinuria after the 20th gestational week in a previously normoten­sive and non protein uric women. According to the National High blood presure Working group (NHBPEP and Ameri­can college of obstetricans and Gynecologiests (ACOG hypertension in pregnancy is defined as a diastolic blood pressure of 90 mm Hg or higher after 20 weeks of gestation in a woman with previously normal blood pressure (NHBPEP, 2000; ACOG, 2002. If the disease is allowed to progress to the HELLP syndrome or eclampsia, maternal morbidity and mortality increases. The majority of perinatal losses are related to placental insufficiency, which causes intrauterine growth retardation, prematurity associated with preterm delivery, or abruptio placentae. Objectives: This study tried to explore the effect of serum C reactive protein concentration in preeclamptic women and its effect on pregnancy outcome.Methods: This case control study included 60 third trimester pregnant women (30 normotensive and 30 preeclamptic who attended Department of Obstetrics and Gynaecology, BIRDEM and DMCH, during July 2009 and June 2010. Estimation of serum C reactive protein (CRP concentrations was done by liquid phase immunoprecipitation assay and turbulometry at DMC.Results: Mean (±SD age showed no significant difference between groups; however, BMI, SBP, DBP and CRP were significantly (P<0.001 high in case group. Gravidity and ANC showed no significant variation between groups. CRP concentration was significantly high case group. Gestational age was significantly low in case group resulting in higher preterm delivery. No significant variation was observed regarding fetal outcome; however, birth weight was significantly low and neonatal complication was also significantly high in case group.Conclusion: CRP concentration was high in

  15. Quantifying sediment source contributions in coastal catchments impacted by the Fukushima nuclear accident with carbon and nitrogen elemental concentrations and stable isotope ratios

    Science.gov (United States)

    Laceby, J. Patrick; Huon Huon, Sylvain; Onda, Yuichi; Evrard, Olivier

    2016-04-01

    The Fukushima Dai-ichi Nuclear Power Plant accidental release of radioactive contaminants resulted in the significant fallout of radiocesium over several coastal catchments in the Fukushima Prefecture. Radiocesium, considered to be the greatest risk to the short and long term health of the local community, is rapidly bound to fine soil particles and thus is mobilized and transported during soil erosion and runoff processes. As there has been a broad-scale decontamination of rice paddy fields and rural residential areas in the contaminated region, one important long term question is whether there is, or may be, a downstream transfer of radiocesium from forests that covered over 65% of the most contaminated region. Accordingly, carbon and nitrogen elemental concentrations and stable isotope ratios are used to determine the relative contributions of forests and rice paddies to transported sediment in three contaminated coastal catchments. Samples were taken from the three main identified sources: cultivated soils (rice paddies and fields, n=30), forest soils (n=45), and subsoils (channel bank and decontaminated soils, n = 25). Lag deposit sediment samples were obtained from five sampling campaigns that targeted the main hydrological events from October 2011 to October 2014. In total, 86 samples of deposited sediment were analyzed for particulate organic matter elemental concentrations and isotope ratios, 24 from the Mano catchment, 44 from the Niida catchment, and 18 from the Ota catchment. Mann-Whitney U-tests were used to examine the source discrimination potential of this tracing suite and select the appropriate tracers for modelling. The discriminant tracers were modelled with a concentration-dependent distribution mixing model. Preliminary results indicate that cultivated sources (predominantly rice paddies) contribute disproportionately more sediment per unit area than forested regions in these contaminated catchments. Future research will examine if there are

  16. Preparation, aroma characteristics and volatile compounds of flavorings from enzymatic hydrolyzed rice bran protein concentrate.

    Science.gov (United States)

    Arsa, Supeeraya; Theerakulkait, Chockchai

    2018-02-19

    Rice bran is a by-product obtained from the rice milling industry. The aims of this research were to add value to rice bran by preparation of enzymatic hydrolyzed rice bran protein concentrate (HRPC) as a flavoring agent and the flavoring which was produced by HRPC has not been investigated. Different drying methods (freeze-drying and spray-drying) and fructose additions were studied for improvement of rice bran protein sensorial aroma characteristics. The most abundant amino acids in liquid HRPC (LH) were glutamic acid, arginine, aspartic acid and leucine. The intensity of desirable aromas, such as cereal-like, nut-like, milk-powder-like, sweet, and cocoa-like aroma, were higher in spray-dried HRPC powder (SHP) than in LH and freeze-dried HRPC. Volatile compounds, such as aldehydes, pyrazines and ketones, were significantly increased in HRPC powders in which fructose was added before spray-drying (SHP-F). Higher amounts of 2-methylbutanal, 3-methylbutanal, phenylacetaldehyde, 2,5-dimethylpyrazine, vanillin, 2-acetylpyrrole and maltol were detected in SHP-F. Moreover, these compounds had high odor active values, which accounted for the cocoa-like, sweet, nut-like, and milk-powder-like characteristics of SHP-F. These findings could lead to the creation of desirable aroma characteristics of rice bran protein concentrate by different preparation methods. © 2018 Society of Chemical Industry. © 2018 Society of Chemical Industry.

  17. EFFECTS OF PROTEIN-XANTHOPHYLL (PX CONCENTRATE OF ALFALFA ADDITIVE TO CRUDE PROTEIN-REDUCED DIETS ON NITROGEN EXCRETION, GROWTH PERFORMANCE AND MEAT QUALITY OF PIGS

    Directory of Open Access Journals (Sweden)

    Eugeniusz GRELA

    2009-06-01

    Full Text Available The infl uence of protein-xanthophyll (PX concentrate of alfalfa supplement to crude protein-reduced diets was examined in relation to nitrogen excretion, performance parameters and pig meat quality. The investigations included 60 growers (PL x PLW x Duroc crossbreeds assigned to 3 groups. The conclusion is that there is a large potential to decrease nitrogen emission to the environment by 10% lowering of dietary crude protein intake along with reduced animal growth rate and elevated mixture utilization. Inclusion of a protein-xanthophyll concentrate (PX of alfalfa to the diet is likely to diminish disadvantageous productive parameters arising from limiting of total crude protein level in relation to the requirements of pigs feeding norms [1993]. At the same time, it improves feed nitrogen utilization and reduces noxious odour emissions from a piggery. The components of a protein-xanthophyll concentrate (PX contribute to increased liver and kidney weight.

  18. Alterations in serum amino acid concentrations in dogs with protein-losing enteropathy.

    Science.gov (United States)

    Kathrani, Aarti; Allenspach, Karin; Fascetti, Andrea J; Larsen, Jennifer A; Hall, Edward J

    2018-03-31

    Certain amino acids are decreased in humans with inflammatory bowel disease (IBD) and supplementation with the same amino acids has shown beneficial effects in animal models of IBD. Currently, the amino acid status of dogs with protein-losing enteropathy (PLE) is unknown. To determine if serum amino acid concentrations are abnormal in dogs with PLE and correlated with clinical and laboratory variables and outcome. Thirty client-owned dogs diagnosed with PLE and 12 apparently healthy dogs seen at Bristol Veterinary School. Retrospective study using stored residual serum from fasted dogs with PLE, collected at the time of diagnostic investigation and from apparently healthy dogs. Serum was analyzed for 30 amino acids using an automated high-performance liquid chromatography amino acid analyzer. Serum tryptophan concentrations were significantly decreased in dogs with PLE (median, 22 nmol/mL; range, 1-80 nmol/mL) compared with apparently healthy control dogs (median, 77.5 nmol/mL; range, 42-135 nmol/mL, P PLE and apparently healthy. Serum tryptophan concentrations were also significantly correlated with serum albumin concentrations in dogs with PLE (P = .001, R 2 = 0.506). Decreased serum tryptophan concentration might play a role in the pathogenesis of canine PLE or be a consequence of the disease. © 2018 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

  19. Influence of different levels of concentrate and ruminally undegraded protein on digestive variables in beef heifers.

    Science.gov (United States)

    Pina, D S; Valadares Filho, S C; Tedeschi, L O; Barbosa, A M; Valadares, R F D

    2009-03-01

    This experiment evaluated the effect of 2 levels of diet concentrate (20 and 40% of DM) and 2 levels of ruminally undegraded protein (RUP: 25 and 40% of CP) on nutrient intake, total and partial apparent nutrient digestibility, microbial protein synthesis, and ruminal and physiological variables. Eight Nellore heifers (233 +/- 14 kg of BW) fitted with ruminal, abomasal, and ileal cannulas were used. The animals were held in individual sheltered pens of approximately 15 m(2) and fed twice daily at 0800 and 1600 h for ad libitum intake. Heifers were allocated in two 4 x 4 Latin square designs, containing 8 heifers, 4 experimental periods, and 4 treatments in a 2 x 2 factorial arrangement. All statistical analyses were performed using PROC MIXED of SAS. Titanium dioxide (TiO(2)) and chromic oxide (Cr(2)O(3)) were used to estimate digesta fluxes and fecal excretion. Purine derivative (PD) excretion and abomasal purine bases were used to estimate the microbial N (MN) synthesis. No significant interaction (P > 0.10) between dietary levels of RUP and concentrate was observed. There was no effect of treatment (P = 0.24) on DMI. Both markers led to the same estimates of fecal, abomasal, and ileal DM fluxes, and digestibilities of DM and individual nutrients. Ruminal pH was affected by sampling time (P RUP, whereas a quadratic effect (P RUP. The higher level of dietary concentrate led to greater MN yield regardless of the level of RUP. The MN yield and the efficiency of microbial yield estimated from urinary PD excretion produced greater (P RUP and concentrate were observed for ruminal and digestive parameters. Neither RUP nor concentrate level affected DMI. Titanium dioxide showed to be similar to Cr(2)O(3) as an external marker to measure digestibility and nutrient fluxes in cattle.

  20. [Thyroid proteins in endemic goitre and their relationship to the intrathyroidal thyroid hormone concentration].

    Science.gov (United States)

    Platzer, S; Groebner, P; Hausen, A; Obendorf, L; Riccabona, G

    1980-02-01

    According to several reports we suspected that the pathogenesis of endemic goitre cannot be explained by iodine deficiency only, but that other--partially endogenous--goitrogenic factors must be present. We therefore studied 16 cases of "euthyroid" endemic goitre from the endemic goitre area of the province of Bolzano in Italy. After fractionation of tissue homogenates, T 4 and T 3 were measured by RIA and the I concentration was also termined. Thyroglobulin and its fractions were measured by ultracentrifuge procedures after assessment of the total protein concentration. Evaluation of the present results suggests that an insufficient synthesis of thyroglobulin in the examined goitres induces an inadequate adaptation of the organism to iodine deficiency, which, in turn, decreases the thyroid hormone concentration in thyroid tissue and enhances goitrogenesis. Considering the normal iodine content of the examined tissues, there obviously seems to be two intrathyroidal iodine pools, one of which supplies the body with thyroid hormones under pituitary stimulation even though its thyroglobulin pool is reduced, while a significant amount of the thyroidal iodine pool is bound in metabolically inert protein molecules and therefore increases the goitrogenic effect of iodine deficiency.

  1. Serum C-reactive protein concentrations in healthy Miniature Schnauzer dogs.

    Science.gov (United States)

    Wong, Valerie M; Kidney, Beverly A; Snead, Elisabeth C R; Myers, Sherry L; Jackson, Marion L

    2011-09-01

    C-reactive protein (CRP) is a sensitive marker for inflammation in people and dogs. In people, an association between CRP concentration and atherosclerosis has been reported. Atherosclerosis is rare in dogs, but the Miniature Schnauzer breed may be at increased risk for developing this vascular disease. It is not known if CRP concentrations in Miniature Schnauzer dogs differ from those in other dog breeds. Our objectives were to validate an automated human CRP assay for measuring CRP in dogs and compare CRP concentrations in healthy Miniature Schnauzer dogs with those in non-Miniature Schnauzer breeds. Sera from 37 non-Miniature Schnauzer dogs with inflammatory disease were pooled and used to validate a human CRP immunoturbidimetric assay for measuring canine CRP. Blood was collected from 20 healthy Miniature Schnauzer dogs and 41 healthy dogs of other breeds. Median serum CRP concentration of healthy Miniature Schnauzer dogs was compared with that of healthy non-Miniature Schnauzer dogs. The human CRP assay measured CRP reliably with linearity between 0 and 20 mg/L. CRP concentration for healthy Miniature Schnauzer dogs (median 4.0 mg/L, minimum-maximum 0-18.2 mg/L) was significantly higher than for the healthy non-Miniature Schnauzer dogs (median 0.1 mg/L, minimum-maximum 0-10.7 mg/L); 17 of the 20 Miniature Schnauzer dogs had values that overlapped with those of the non-Miniature Schnauzer dogs. Median CRP concentration of Miniature Schnauzer dogs was slightly higher than that of other breeds of dogs. A relationship between higher CRP concentration in Miniature Schnauzer dogs and idiopathic hyperlipidemia, pancreatitis, and possible increased risk for atherosclerosis remains to be determined. ©2011 American Society for Veterinary Clinical Pathology.

  2. Processing traits and digestibility of extruded dog foods with soy protein concentrate.

    Science.gov (United States)

    Venturini, K S; Sarcinelli, M F; Baller, M A; Putarov, T C; Malheiros, E B; Carciofi, A C

    2018-04-11

    Soya bean protein concentrate (SPC) with two particle sizes were evaluated on extrusion parameters, kibble formation, digestibility and palatability of dog foods. Eight diets were extruded: PBM-control diet based on poultry by-product meal (PBM); GM-a diet in which corn gluten meal (GM) replaced 45% of the diet protein; cSPC15%, cSPC30% and cSPC45%-diets in which SPC of coarse particle size (600 μm) replaced 15%, 30% and 45% of the diet protein; and sSPC15%, sSPC30% and sSPC45%-diets in which SPC of small particle size (200 μm) replaced 15%, 30% and 45% of the diet protein. The digestibility of nutrients was evaluated for the PBM, GM, cSPC45% and sSPC45% diets, using six dogs per food. The PBM, GM and cSPC45% diets were compared for palatability. Data were submitted for analysis of variance, and the means were compared by polynomial contrasts or Tukey's test (p beans resulted in an ingredient with low fermentable fibre content, which did not alter faecal formation or characteristics. © 2018 Blackwell Verlag GmbH.

  3. A Graphical User Interface for Software-assisted Tracking of Protein Concentration in Dynamic Cellular Protrusions.

    Science.gov (United States)

    Saha, Tanumoy; Rathmann, Isabel; Galic, Milos

    2017-07-11

    Filopodia are dynamic, finger-like cellular protrusions associated with migration and cell-cell communication. In order to better understand the complex signaling mechanisms underlying filopodial initiation, elongation and subsequent stabilization or retraction, it is crucial to determine the spatio-temporal protein activity in these dynamic structures. To analyze protein function in filopodia, we recently developed a semi-automated tracking algorithm that adapts to filopodial shape-changes, thus allowing parallel analysis of protrusion dynamics and relative protein concentration along the whole filopodial length. Here, we present a detailed step-by-step protocol for optimized cell handling, image acquisition and software analysis. We further provide instructions for the use of optional features during image analysis and data representation, as well as troubleshooting guidelines for all critical steps along the way. Finally, we also include a comparison of the described image analysis software with other programs available for filopodia quantification. Together, the presented protocol provides a framework for accurate analysis of protein dynamics in filopodial protrusions using image analysis software.

  4. The structure of mAG, a monomeric mutant of the green fluorescent protein Azami-Green, reveals the structural basis of its stable green emission

    International Nuclear Information System (INIS)

    Ebisawa, Tatsuki; Yamamura, Akihiro; Kameda, Yasuhiro; Hayakawa, Kou; Nagata, Koji; Tanokura, Masaru

    2010-01-01

    The crystal structure of a monomeric mutant of Azami-Green (mAG) from G. fascicularis was determined at 2.2 Å resolution. Monomeric Azami-Green (mAG) from the stony coral Galaxea fascicularis is the first known monomeric green-emitting fluorescent protein that is not a variant of Aequorea victoria green fluorescent protein (avGFP). These two green fluorescent proteins are only 27% identical in their amino-acid sequences. mAG is more similar in its amino-acid sequence to four fluorescent proteins: Dendra2 (a green-to-red irreversibly photoconverting fluorescent protein), Dronpa (a bright-and-dark reversibly photoswitchable fluorescent protein), KikG (a tetrameric green-emitting fluorescent protein) and Kaede (another green-to-red irreversibly photoconverting fluorescent protein). To reveal the structural basis of stable green emission by mAG, the 2.2 Å crystal structure of mAG has been determined and compared with the crystal structures of avGFP, Dronpa, Dendra2, Kaede and KikG. The structural comparison revealed that the chromophore formed by Gln62-Tyr63-Gly64 (QYG) and the fixing of the conformation of the imidazole ring of His193 by hydrogen bonds and van der Waals contacts involving His193, Arg66 and Thr69 are likely to be required for the stable green emission of mAG. The crystal structure of mAG will contribute to the design and development of new monomeric fluorescent proteins with faster maturation, brighter fluorescence, improved photostability, new colours and other preferable properties as alternatives to avGFP and its variants

  5. Annotating N termini for the human proteome project: N termini and Nα-acetylation status differentiate stable cleaved protein species from degradation remnants in the human erythrocyte proteome.

    Science.gov (United States)

    Lange, Philipp F; Huesgen, Pitter F; Nguyen, Karen; Overall, Christopher M

    2014-04-04

    A goal of the Chromosome-centric Human Proteome Project is to identify all human protein species. With 3844 proteins annotated as "missing", this is challenging. Moreover, proteolytic processing generates new protein species with characteristic neo-N termini that are frequently accompanied by altered half-lives, function, interactions, and location. Enucleated and largely void of internal membranes and organelles, erythrocytes are simple yet proteomically challenging cells due to the high hemoglobin content and wide dynamic range of protein concentrations that impedes protein identification. Using the N-terminomics procedure TAILS, we identified 1369 human erythrocyte natural and neo-N-termini and 1234 proteins. Multiple semitryptic N-terminal peptides exhibited improved mass spectrometric identification properties versus the intact tryptic peptide enabling identification of 281 novel erythrocyte proteins and six missing proteins identified for the first time in the human proteome. With an improved bioinformatics workflow, we developed a new classification system and the Terminus Cluster Score. Thereby we described a new stabilizing N-end rule for processed protein termini, which discriminates novel protein species from degradation remnants, and identified protein domain hot spots susceptible to cleavage. Strikingly, 68% of the N-termini were within genome-encoded protein sequences, revealing alternative translation initiation sites, pervasive endoproteolytic processing, and stabilization of protein fragments in vivo. The mass spectrometry proteomics data have been deposited to ProteomeXchange with the data set identifier .

  6. Single-step transesterification with simultaneous concentration and stable isotope analysis of fatty acid methyl esters by gas chromatography-combustion-isotope ratio mass spectrometry.

    Science.gov (United States)

    Panetta, Robert J; Jahren, A Hope

    2011-05-30

    Gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS) is increasingly applied to food and metabolic studies for stable isotope analysis (δ(13) C), with the quantification of analyte concentration often obtained via a second alternative method. We describe a rapid direct transesterification of triacylglycerides (TAGs) for fatty acid methyl ester (FAME) analysis by GC-C-IRMS demonstrating robust simultaneous quantification of amount of analyte (mean r(2) =0.99, accuracy ±2% for 37 FAMEs) and δ(13) C (±0.13‰) in a single analytical run. The maximum FAME yield and optimal δ(13) C values are obtained by derivatizing with 10% (v/v) acetyl chloride in methanol for 1 h, while lower levels of acetyl chloride and shorter reaction times skewed the δ(13) C values by as much as 0.80‰. A Bland-Altman evaluation of the GC-C-IRMS measurements resulted in excellent agreement for pure oils (±0.08‰) and oils extracted from French fries (±0.49‰), demonstrating reliable simultaneous quantification of FAME concentration and δ(13) C values. Thus, we conclude that for studies requiring both the quantification of analyte and δ(13) C data, such as authentication or metabolic flux studies, GC-C-IRMS can be used as the sole analytical method. Copyright © 2011 John Wiley & Sons, Ltd.

  7. Nonlinear concentration gradients regulated by the width of channels for observation of half maximal inhibitory concentration (IC50) of transporter proteins.

    Science.gov (United States)

    Abe, Yuta; Kamiya, Koki; Osaki, Toshihisa; Sasaki, Hirotaka; Kawano, Ryuji; Miki, Norihisa; Takeuchi, Shoji

    2015-08-21

    This paper describes a simple microfluidic device that can generate nonlinear concentration gradients. We changed the "width" of channels that can drastically shorten the total microfluidic channel length and simplify the microfluidic network design rather than the "length" of channels. The logarithmic concentration gradients generated by the device were in good agreement with those obtained by simulation. Using this device, we evaluated a probable IC50 value of the ABC transporter proteins by the competitive transport assays at five different logarithmic concentrations. This probable IC50 value was in good agreement with an IC50 value (0.92 μM) obtained at the diluted concentrations of seven points.

  8. Protein intercalation in DNA as one of main modes of fixation of the most stable chromatin loop domains

    Directory of Open Access Journals (Sweden)

    М. I. Chopei

    2014-08-01

    Full Text Available The main mechanism of DNA track formation during comet assay of nucleoids, obtained after removal of cell membranes and most of proteins, is the extension to anode of negatively supercoiled DNA loops attached to proteins, remaining in nucleoid after lysis treatment. The composition of these residual protein structures and the nature of their strong interaction with the loop ends remain poorly studied. In this work we investigated the influence of chloroquine intercalation and denaturation of nucleoid proteins on the efficiency of electrophoretic track formation during comet assay. The results obtained suggest that even gentle protein denaturation is sufficient to reduce considerably the effectiveness of the DNA loop migration due to an increase in the loops size. The same effect was observed under local DNA unwinding upon chloroquine intercalation around the sites of the attachment of DNA to proteins. The topological interaction (protein intercalation into the double helix between DNA loop ends and nucleoid proteins is discussed.

  9. Isoflavonas em isolados e concentrados protéicos de soja Isoflavones in soy protein isolate and soy protein concentrate

    Directory of Open Access Journals (Sweden)

    Maria Cristina Y. Lui

    2003-12-01

    Full Text Available Isolados e concentrados protéicos de soja são ingredientes largamente utilizados na indústria de panificação, confeitaria, bebidas e embutidos. As isoflavonas presentes na soja podem sofrer alterações em quantidade e perfil de distribuição dependendo das condições de processamento. O objetivo deste estudo foi verificar o balanço de massa de isoflavonas e proteína em processamento de isolados e de concentrados protéicos de soja (tratamento com ácido e com álcool. A maior parte das isoflavonas presentes na matéria-prima (farinha desengordurada de soja é perdida nos sobrenadantes de processo (90% para extração com etanol 60%, 52% para processamento de isolado protéico e 47% para extração com ácido. O teor de isoflavonas nos produtos obtidos foi de 686µg/g base seca (b.s. para isolado protéico, 871µ g/g b.s. para concentrado protéico obtido por tratamento ácido e apenas 153µg/g b.s. para concentrado protéico obtido por tratamento com álcool. Não foi observada alteração no perfil de distribuição das isoflavonas nesse último processo, enquanto que nos dois primeiros notou-se diminuição da quantidade das formas malonil glicosídeos e aumento da quantidade das formas beta-glicosiladas e gliconas.Soy protein isolate (SPI and soy protein concentrate (SPC are largely used in bakery, confectionary, meat and beverage products. Isoflavones present in soybeans products can undergo changes in quantity and profile depending on the processing conditions. The objective of this work was to conduct mass balance studies of isoflavones and protein during the processing of SPI and SPC (acid and alcohol leach. The majority of isoflavones present in the raw material is lost in the supernatants (90% for SPC treated with alcohol, 52% for SPI and 47% for SPC treated with acid. Total concentration of isoflavones was 652µg/g for SPI, 838µg/g for SPC (acid leach, and only 147µg/g for SPC (alcohol leach. There were no changes in the

  10. Deciphering Systemic Wound Responses of the Pumpkin Extrafascicular Phloem by Metabolomics and Stable Isotope-Coded Protein Labeling1[C][W

    Science.gov (United States)

    Gaupels, Frank; Sarioglu, Hakan; Beckmann, Manfred; Hause, Bettina; Spannagl, Manuel; Draper, John; Lindermayr, Christian; Durner, Jörg

    2012-01-01

    In cucurbits, phloem latex exudes from cut sieve tubes of the extrafascicular phloem (EFP), serving in defense against herbivores. We analyzed inducible defense mechanisms in the EFP of pumpkin (Cucurbita maxima) after leaf damage. As an early systemic response, wounding elicited transient accumulation of jasmonates and a decrease in exudation probably due to partial sieve tube occlusion by callose. The energy status of the EFP was enhanced as indicated by increased levels of ATP, phosphate, and intermediates of the citric acid cycle. Gas chromatography coupled to mass spectrometry also revealed that sucrose transport, gluconeogenesis/glycolysis, and amino acid metabolism were up-regulated after wounding. Combining ProteoMiner technology for the enrichment of low-abundance proteins with stable isotope-coded protein labeling, we identified 51 wound-regulated phloem proteins. Two Sucrose-Nonfermenting1-related protein kinases and a 32-kD 14-3-3 protein are candidate central regulators of stress metabolism in the EFP. Other proteins, such as the Silverleaf Whitefly-Induced Protein1, Mitogen Activated Protein Kinase6, and Heat Shock Protein81, have known defensive functions. Isotope-coded protein labeling and western-blot analyses indicated that Cyclophilin18 is a reliable marker for stress responses of the EFP. As a hint toward the induction of redox signaling, we have observed delayed oxidation-triggered polymerization of the major Phloem Protein1 (PP1) and PP2, which correlated with a decline in carbonylation of PP2. In sum, wounding triggered transient sieve tube occlusion, enhanced energy metabolism, and accumulation of defense-related proteins in the pumpkin EFP. The systemic wound response was mediated by jasmonate and redox signaling. PMID:23085839

  11. Absolute quantitative autoradiography of low concentrations of [125I]-labeled proteins in arterial tissue

    International Nuclear Information System (INIS)

    Schnitzer, J.J.; Morrel, E.M.; Colton, C.K.; Smith, K.A.; Stemerman, M.B.

    1987-01-01

    We developed a method for absolute quantitative autoradiographic measurement of very low concentrations of [ 125 I]-labeled proteins in arterial tissue using Kodak NTB-2 nuclear emulsion. A precise linear relationship between measured silver grain density and isotope concentration was obtained with uniformly labeled standard sources composed of epoxy-embedded gelatin containing glutaraldehyde-fixed [ 125 I]-albumin. For up to 308-day exposures of 1 micron-thick tissue sections, background grain densities ranged from about two to eight grains/1000 micron 2, and the technique was sensitive to as little as about one grain/1000 micron 2 above background, which correspond to a radioactivity concentration of about 2 x 10(4) cpm/ml. A detailed statistical analysis of variability was performed and the sum of all sources of variation quantified. The half distance for spatial resolution was 1.7 micron. Both visual and automated techniques were employed for quantitative grain density analysis. The method was illustrated by measurement of in vivo transmural [ 125 I]-low-density lipoprotein [( 125 I]-LDL) concentration profiles in de-endothelialized rabbit thoracic aortic wall

  12. [Examination of acute phase proteins concentrations in children with allergic rhinitis].

    Science.gov (United States)

    Steiner, Iwona; Sobieska, Magdalena; Pucher, Beata; Grzegorowski, Michał; Samborski, Włodzimierz

    2006-01-01

    , blood sample was taken after written parents' consent. In all children skin tests (Stallergens) were performed at the beginning of the year. Any reaction was present in the control group. Following proteins were analyzed: CRP, AGP, alpha1-antichymotrypsin (ACT), transferrin, ceruloplasmin (Cp), alpha1-antitrypsin, haptoglobin and alpha2-macroglobulin (A2M). C-reactive protein level was very low, what allowed us to exclude all acute conditions. As expected, AGP and ACT concentrations were not elevated, either, and even non significantly lower values were observed in allergic children in comparison to controls. It is worth noticing that transferrin concentration was statistically lower in allergic children, as well as A2M and Cp concentrations. All this suggest an on-going disturbance in cytokine network that may directly affect both the iron metabolism and the non-specific immunity. It may be stated that allergic rhinitis causes impairment of acute phase proteins synthesis, which may affect natural defense or homeostasis in the immune system of a child.

  13. Interaction between Nbp35 and Cfd1 proteins of cytosolic Fe-S cluster assembly reveals a stable complex formation in Entamoeba histolytica.

    Directory of Open Access Journals (Sweden)

    Shadab Anwar

    Full Text Available Iron-Sulfur (Fe-S proteins are involved in many biological functions such as electron transport, photosynthesis, regulation of gene expression and enzymatic activities. Biosynthesis and transfer of Fe-S clusters depend on Fe-S clusters assembly processes such as ISC, SUF, NIF, and CIA systems. Unlike other eukaryotes which possess ISC and CIA systems, amitochondriate Entamoeba histolytica has retained NIF & CIA systems for Fe-S cluster assembly in the cytosol. In the present study, we have elucidated interaction between two proteins of E. histolytica CIA system, Cytosolic Fe-S cluster deficient 1 (Cfd1 protein and Nucleotide binding protein 35 (Nbp35. In-silico analysis showed that structural regions ranging from amino acid residues (P33-K35, G131-V135 and I147-E151 of Nbp35 and (G5-V6, M34-D39 and G46-A52 of Cfd1 are involved in the formation of protein-protein complex. Furthermore, Molecular dynamic (MD simulations study suggested that hydrophobic forces surpass over hydrophilic forces between Nbp35 and Cfd1 and Van-der-Waal interaction plays crucial role in the formation of stable complex. Both proteins were separately cloned, expressed as recombinant fusion proteins in E. coli and purified to homogeneity by affinity column chromatography. Physical interaction between Nbp35 and Cfd1 proteins was confirmed in vitro by co-purification of recombinant Nbp35 with thrombin digested Cfd1 and in vivo by pull down assay and immunoprecipitation. The insilico, in vitro as well as in vivo results prove a stable interaction between these two proteins, supporting the possibility of its involvement in Fe-S cluster transfer to target apo-proteins through CIA machinery in E. histolytica. Our study indicates that initial synthesis of a Fe-S precursor in mitochondria is not necessary for the formation of Cfd1-Nbp35 complex. Thus, Cfd1 and Nbp35 with the help of cytosolic NifS and NifU proteins can participate in the maturation of non-mitosomal Fe-S proteins

  14. Concentration of rat brown adipose tissue uncoupling protein may not be correlated with 3H-GDP binding

    International Nuclear Information System (INIS)

    Henningfield, M.F.; Swick, A.G.; Swick, R.W.

    1986-01-01

    Rats fed diets low in protein or exposed to cold show an increase in brown adipose tissue (BAT) mitochondrial 3 H-GDP binding. To investigate this phenomenon further, the uncoupling protein associated with BAT function was measured immunochemically on nitrocellulose blots. Quantitation of uncoupling protein was achieved by densitometer scanning with a BioRad densitometer. Peaks were integrated with Chromatochart software and an Apple IIe computer. A standard curve of purified uncoupling protein (50 to 500 ng) was used to calculate uncoupling protein concentration. There is a 1.5-fold increase in uncoupling protein per mg of protein in BAT mitochondria from rats exposed to cold for 15 days. There was no decrease in uncoupling protein from rats exposed to the cold followed by 24 h at 27 0 C although 3 H-GDP binding had decreased by half. Rats fed diets containing either 5 or 15% lactalbumin for 3 weeks did not show differences in uncoupling protein concentration although 3 H-GDP binding was 1.5-fold greater in BAT mitochondria from the low protein group. These results indicate that GDP binding does not necessarily reflect the concentration of uncoupling protein in BAT mitochondria

  15. Production of Protein Concentrate and 1,3-Propanediol by Wheat-Based Thin Stillage Fermentation.

    Science.gov (United States)

    Ratanapariyanuch, Kornsulee; Shim, Youn Young; Emami, Shahram; Reaney, Martin J T

    2017-05-17

    Fermentation of wheat with yeast produces thin stillage (W-TS) and distiller's wet grains. A subsequent fermentation of W-TS (two-stage fermentation, TSF) with endemic bacteria at 25 and 37 °C decreased glycerol and lactic acid concentrations, while 1,3-propanediol (1,3-PD) and acetic acid accumulated with greater 1,3-PD and acetic acid produced at 37 °C. During TSF, W-TS colloids coagulated and floated in the fermentation medium producing separable liquid and slurry fractions. The predominant endemic bacteria in W-TS were Lactobacillus panis, L. gallinarum, and L. helveticus, and this makeup did not change substantially as fermentation progressed. As nutrients were exhausted, floating particles precipitated. Protein contents of slurry and clarified liquid increased and decreased, respectively, as TSF progressed. The liquid was easily filtered through an ultrafiltration membrane. These results suggested that TSF is a novel method for W-TS clarification and production of protein concentrates and 1,3-PD from W-TS.

  16. Quantification of protein concentration by the Bradford method in the presence of pharmaceutical polymers.

    Science.gov (United States)

    Carlsson, Nils; Borde, Annika; Wölfel, Sebastian; Kerman, Björn; Larsson, Anette

    2011-04-01

    We investigated how the Bradford assay for measurements of protein released from a drug formulation may be affected by a concomitant release of a pharmaceutical polymer used to formulate the protein delivery device. The main result is that polymer-caused perturbations of the Coomassie dye absorbance at the Bradford monitoring wavelength (595nm) can be identified and corrected by recording absorption spectra in the region of 350-850mm. The pharmaceutical polymers Carbopol and chitosan illustrate two potential types of perturbations in the Bradford assay, whereas the third polymer, hydroxypropylmethylcellulose (HPMC), acts as a nonperturbing control. Carbopol increases the apparent absorbance at 595nm because the polymer aggregates at the low pH of the Bradford protocol, causing a turbidity contribution that can be corrected quantitatively at 595nm by measuring the sample absorbance at 850nm outside the dye absorption band. Chitosan is a cationic polymer under Bradford conditions and interacts directly with the anionic Coomassie dye and perturbs its absorption spectrum, including 595nm. In this case, the Bradford method remains useful if the polymer concentration is known but should be used with caution in release studies where the polymer concentration may vary and needs to be measured independently. Copyright © 2010 Elsevier Inc. All rights reserved.

  17. Effects of diets containing vegetable protein concentrates on performance and activity of digestive enzymes in silver catfish (Rhamdia quelen

    Directory of Open Access Journals (Sweden)

    Naglezi de Menezes Lovatto

    2014-02-01

    Full Text Available The purpose of study was to evaluate the effect of using protein concentrates crambe and sunflower meal in the diet of silver catfish juveniles, as substitute for animal protein source. A total of 300 silver catfish had been separate in 15 experimental units of 280 L, totaling five treatments with three replications. We evaluated two levels (25% and 50% replacement of the meat and bone meal by protein concentrates of crambe and sunflower meals. Evaluated growth parameters, biological index and digestive enzymes in fish. There was no statistical difference for mass (g and standard length (cm, but the fish diet CPFCr-25% had greater total length (cm. No difference in dry matter, crude protein and total protein deposited (calculated. However, there was a higher concentration of ash in the carcass of the animals fed the control diet and CPFCr-50% in relation to diet CPFG- 50%, in addition, higher levels of lipids in fish fed diet CPFG-50%. No significant differences for hepatosomatic index, digestive somatic index and intestinal quotient of animals subjected to different treatments. The activity of digestive enzymes trypsin and chymotrypsin did not change. There was increased activity of acid protease. The quantitative and qualitative increase in protein concentration from this fraction allows the use of bran protein concentrates crambe and sunflower as substitutes for animal protein source.

  18. Technology development of protein rich concentrates for nutrition in extreme conditions using soybean and meat by-products.

    Science.gov (United States)

    Kalenik, Tatiana K; Costa, Rui; Motkina, Elena V; Kosenko, Tamara A; Skripko, Olga V; Kadnikova, Irina A

    2017-01-01

    There is a need to develop new foods for participants of expeditions in extreme conditions, which must be self-sufficient. These foods should be light to carry, with a long shelf life, tasty and with  high nutrient density. Currently, protein sources are limited mainly to dried and canned meat. In this work, a protein-rich dried concentrate suitable for extreme expeditions was developed using soya, tomato, milk whey and meat by-products. Protein concentrates were developed using minced beef liver and heart, dehydrated and mixed with a soya protein-lycopene coagulate (SPLC) obtained from a solution prepared with germi- nated soybeans and mixed with tomato paste in milk whey, and finally dried. The technological parameters of pressing SPLC and of drying the protein concentrate were optimized using response surface methodology. The optimized technological parameters to prepare the protein concentrates were obtained, with 70:30 being the ideal ratio of minced meat to SPLC. The developed protein concentrates are characterized by a high calorific value of 376 kcal/100 g of dry product, with a water content of 98 g·kg-1, and 641-644 g·kg-1 of proteins. The essential amino acid indices are 100, with minimum essential amino acid content constitut- ing 100-128% of the FAO standard, depending on the raw meat used. These concentrates are also rich in micronutrients such as β-carotene and vitamin C. Analysis of the nutrient content showed that these non-perishable concentrates present a high nutritional value and complement other widely available vegetable concentrates to prepare a two-course meal. The soups and porridges prepared with these concentrates can be classified as functional foods, and comply with army requirements applicable to food products for extreme conditions.

  19. Microbial protein synthesis and concentration of urea in dairy heifers fed diets with cactus forage Opuntia

    Directory of Open Access Journals (Sweden)

    Maria do Socorro Mercês Aguiar

    2015-04-01

    Full Text Available The objective was to analyze the influence of increasing levels of forage cactus Opuntia in the diet on the nitrogen balance, the concentrations of urea in urine and plasma and microbial protein synthesis in dairy heifers ¾ Holstein-zebu confined. twenty four heifers were used with initial body weight of 163.00 ± 18 kg, with 8 months old and distributed in a completely randomized design with four treatments and six replications. It was used sorghum silage, concentrate and increasing levels of forage cactus Opuntia in the diet (0, 200, 400 and 600 g kg-1. The nitrogen intake, feces and urine, digested and retained with the addition of forage cactus in the diet showed decreasing linear effect. Nitrogen balance was influenced by the inclusion of forage cactus in the diet of dairy heifers through the values observed for the digested and retained nitrogen, which can be related to similar effects found for the consumption of nitrogen and the nitrogen excretion in feces and urine. Nitrogen digested percentage of intake and nitrogen retention as a percentage of ingested and digested showed no difference with the inclusion of forage cactus in the diet. The concentration of urea nitrogen in the urine of heifers had a quadratic effect point of maximum excretion level of 275.80 g kg-1 of forage cactus in the diet. Consequently, the excretion of urea nitrogen and urea excretion showed similar effect with maximum points excretion levels of 293.75 and 319.00 g kg-1 of forage in the diet. The concentration of ureic nitrogen in plasma showed no difference, with an average value of 13.19 mg dL-1. Synthesis of nitrogen and microbial crude protein adjusted to the quadratic model. The microbial efficiency was not influenced by the inclusion of forage cactus in replacement of sorghum silage and concentrate. The urine volume similar to the treatments, with an average of 5.90 liters of urine per day, proving that the creatinine excretion in urine was not influenced

  20. Visfatin and retinol-binding protein 4 concentrations in lean, glucose-tolerant women with PCOS.

    Science.gov (United States)

    Yildiz, Bulent O; Bozdag, Gurkan; Otegen, Umit; Harmanci, Ayla; Boynukalin, Kubra; Vural, Zehra; Kirazli, Serafettin; Yarali, Hakan

    2010-01-01

    Since insulin resistance is accepted to be a common feature of polycystic ovary syndrome (PCOS), the exact molecular mechanism(s) involved in glucose and lipid metabolism have been under investigation in the syndrome. Recently, two novel adipokines, namely visfatin and retinol-binding protein 4 (RBP4), have been suggested to play a role in insulin resistance and diabetes. This study sought to determine whether plasma concentrations of visfatin and RBP4 are altered in PCOS by comparing a total of 27 lean, normal glucose-tolerant PCOS patients with 19 age- and body mass index-matched healthy controls. The mean plasma visfatin concentrations were higher in PCOS patients than those in healthy subjects (37.9+/-18.2 versus 19.8+/-17.5, PPCOS (r=0.52, Plean, glucose-tolerant women with PCOS have increased circulating visfatin and unaltered RBP4 concentrations compared with healthy lean women. In order to clarify overlapping effects and their potential contribution to the pathophysiology of PCOS, further studies are needed. Copyright (c) 2009 Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

  1. Synthesis of stable nanosilver particles (AgNPs) by the proteins of seagrass Syringodium isoetifolium and its biomedicinal properties

    Digital Repository Service at National Institute of Oceanography (India)

    Ahila, N.K.; Ramkumar, V.S.; Prakash, S.; Manikandan, B.; Ravindran, J.; Dhanalakshmi, P.K.; Kannapirana, E.

    A simple eco-friendly approach for the hasty synthesis of stable, potent and benign silver nanoparticles (AgNPs) using seagrass, Syringodium isoetifolium was proposed and described here. The UV–Vis, DLS, XRD, AFM, FESEM, EDX and HRTEM analysis...

  2. Optimization of the protein concentration process from residual peanut oil-cake

    Directory of Open Access Journals (Sweden)

    Gayol, M. F.

    2013-12-01

    Full Text Available The objective of this study was to find the best process conditions for preparing protein concentrate from residual peanut oil-cake (POC. The study was carried out on POC from industrial peanut oil extraction. Different protein extraction and precipitation conditions were used: water/ flour ratio (10:1, 20:1 and 30:1, pH (8, 9 and 10, NaCl concentration (0 and 0.5 M, extraction time (30, 60 and 120 min, temperature (25, 40 and 60 °C, extraction stages (1, 2 and 3, and precipitation pH (4, 4.5 and 5. The extraction and precipitation conditions which showed the highest protein yield were 10:1 water/flour ratio, extraction at pH 9, no NaCl, 2 extraction stages of 30 min at 40 °C and precipitation at pH 4.5. Under these conditions, the peanut protein concentrate (PC contained 86.22% protein, while the initial POC had 38.04% . POC is an alternative source of protein that can be used for human consumption or animal nutrition. Therefore, it adds value to an industry residue.El objetivo de este trabajo fue encontrar las mejores condiciones para obtener un concentrado de proteínas a partir de la torta residual de maní (POC. El estudio se llevó a cabo en POC provenientes de la extracción industrial de aceite de maní. Se utilizaron distintas condiciones para la extracción y precipitación de proteínas: relación agua / harina (10:1, 20:1 y 30:1, pH de extracción (8, 9 y 10, concentración de NaCl (0 y 0,5 M, tiempo de extracción (30, 60 y 120 min, temperatura (25, 40 y 60 °C, número de etapas de extracción (1, 2 y 3, y el pH de precipitación (4, 4,5 y 5. Las condiciones de extracción y de precipitación que mostraron mayor rendimiento de proteína fueron: relación de 10:1 en agua / harina, pH de extracción de 9, en ausencia de NaCl, 2 etapas de extracción de 30 min cada una a 40 °C y el pH de precipitación de 4,5. En estas condiciones, el concentrado de proteína de maní (PC fue de 86,22%, mientras que el porcentaje de proteínas de

  3. Determination of protein concentration in raw milk by mid-infrared fourier transform infrared/attenuated total reflectance spectroscopy.

    Science.gov (United States)

    Etzion, Y; Linker, R; Cogan, U; Shmulevich, I

    2004-09-01

    This study investigates the potential use of attenuated total reflectance spectroscopy in the mid-infrared range for determining protein concentration in raw cow milk. The determination of protein concentration is based on the characteristic absorbance of milk proteins, which includes 2 absorbance bands in the 1500 to 1700 cm(-1) range, known as the amide I and amide II bands, and absorbance in the 1060 to 1100 cm(-1) range, which is associated with phosphate groups covalently bound to casein proteins. To minimize the influence of the strong water band (centered around 1640 cm(-1)) that overlaps with the amide I and amide II bands, an optimized automatic procedure for accurate water subtraction was applied. Following water subtraction, the spectra were analyzed by 3 methods, namely simple band integration, partial least squares (PLS) and neural networks. For the neural network models, the spectra were first decomposed by principal component analysis (PCA), and the neural network inputs were the spectra principal components scores. In addition, the concentrations of 2 constituents expected to interact with the protein (i.e., fat and lactose) were also used as inputs. These approaches were tested with 235 spectra of standardized raw milk samples, corresponding to 26 protein concentrations in the 2.47 to 3.90% (weight per volume) range. The simple integration method led to very poor results, whereas PLS resulted in prediction errors of about 0.22% protein. The neural network approach led to prediction errors of 0.20% protein when based on PCA scores only, and 0.08% protein when lactose and fat concentrations were also included in the model. These results indicate the potential usefulness of Fourier transform infrared/attenuated total reflectance spectroscopy for rapid, possibly online, determination of protein concentration in raw milk.

  4. Impact of Ovine Whey Protein Concentrates and Clarification By-Products on the Yield and Quality of Whey Cheese

    OpenAIRE

    Carlos D. Pereira; Olga Díaz; Angel Cobos

    2007-01-01

    The effects of the addition of whey protein concentrates and clarification by-products obtained from ovine cheese whey and deproteinized whey (Sorelho) on the yield and quality of the whey cheese (Requeijão) have been evaluated. Whey protein concentrates were obtained by ultrafiltration of skimmed whey and Sorelho. The clarification by-products were obtained after the treatment of the skimmed whey and Sorelho by thermocalcic precipitation and microfiltration with two membranes (0.20 and 0.65 ...

  5. Analysis of Low Frequency Protein Truncating Stop-Codon Variants and Fasting Concentration of Growth Hormone.

    Directory of Open Access Journals (Sweden)

    Erik Hallengren

    Full Text Available The genetic background of Growth Hormone (GH secretion is not well understood. Mutations giving rise to a stop codon have a high likelihood of affecting protein function.To analyze likely functional stop codon mutations that are associated with fasting plasma concentration of Growth Hormone.We analyzed stop codon mutations in 5451 individuals in the Malmö Diet and Cancer study by genotyping the Illumina Exome Chip. To enrich for stop codon mutations with likely functional effects on protein function, we focused on those disrupting >80% of the predicted amino acid sequence, which were carried by ≥ 10 individuals. Such mutations were related to GH concentration, measured with a high sensitivity assay (hs-GH and, if nominally significant, to GH related phenotypes, using linear regression analysis.Two stop codon mutations were associated with the fasting concentration of hs-GH. rs121909305 (NP_005370.1:p.R93* [Minor Allele Frequency (MAF = 0.8%] in the Myosin 1A gene (MYO1A was associated with a 0.36 (95%CI, 0.04 to 0.54; p=0.02 increment of the standardized value of the natural logarithm of hs-GH per 1 minor allele and rs35699176 (NP_067040.1:p.Q100* in the Zink Finger protein 77 gene (ZNF77 (MAF = 4.8% was associated with a 0.12 (95%CI, 0.02 to 0.22; p = 0.02 increase of hs-GH. The mutated high hs-GH associated allele of MYO1A was related to lower BMI (β-coefficient, -0.22; p = 0.05, waist (β-coefficient, -0.22; p = 0.04, body fat percentage (β-coefficient, -0.23; p = 0.03 and with higher HDL (β-coefficient, 0.23; p = 0.04. The ZNF77 stop codon was associated with height (β-coefficient, 0.11; p = 0.02 but not with cardiometabolic risk factors.We here suggest that a stop codon of MYO1A, disrupting 91% of the predicted amino acid sequence, is associated with higher hs-GH and GH-related traits suggesting that MYO1A is involved in GH metabolism and possibly body fat distribution. However, our results are preliminary and need replication in

  6. High levels of stable p53 protein and the expression of c-myc in cultured human epithelial tissue after cobalt-60 irradiation

    International Nuclear Information System (INIS)

    Mothersill, C.; Seymour, C.B.; Harney, J.; Hennessy, T.P.

    1994-01-01

    When explants of human uroepithelium or esophageal epithelium are exposed to acute doses of radiation (cobalt-60), the cells which grow out to form the primary cultures show a number of abnormal features. These include the development of characteristic nonsenescent foci. These foci have previously been shown to be c-myc positive and to have an abnormal, tumor-like ultrastructure. Expression of c-myc and the level of stable p53 proteins have now been examined in these cultures 2 weeks after irradiation. Both proteins occurred in dividing cells at the growing edge of the explant and in the foci. The expression of c-myc appeared to be correlated with growth. As expected, variation between individual cultures of normal human cells was noted in the expression of stable p53 protein. Most control uroepithelial cell cultures were negative, but a small cohort showed a wide range of values. The control cultures from the esophageal tissues had high expression of p53, and this decreased marginally after irradiation. Cells positive for p53 were always in cycle and were usually positive for c-myc as well. It would appear from these results that the expression of c-myc and the stable form of the p53 protein occur in irradiated primary cultures of normal human cells both in foci which also express a number of abnormalities and in open-quotes edgeclose quotes cells which are dividing. Cultures of unirradiated cells from esophagus and a small number of uroepithelial samples had high levels of p53. Possible reasons for this are discussed. 33 refs., 2 figs., 3 tabs

  7. Baseline Plasma C-Reactive Protein Concentrations and Motor Prognosis in Parkinson Disease.

    Directory of Open Access Journals (Sweden)

    Atsushi Umemura

    Full Text Available C-reactive protein (CRP, a blood inflammatory biomarker, is associated with the development of Alzheimer disease. In animal models of Parkinson disease (PD, systemic inflammatory stimuli can promote neuroinflammation and accelerate dopaminergic neurodegeneration. However, the association between long-term systemic inflammations and neurodegeneration has not been assessed in PD patients.To investigate the longitudinal effects of baseline CRP concentrations on motor prognosis in PD.Retrospective analysis of 375 patients (mean age, 69.3 years; mean PD duration, 6.6 years. Plasma concentrations of high-sensitivity CRP were measured in the absence of infections, and the Unified Parkinson's Disease Rating Scale Part III (UPDRS-III scores were measured at five follow-up intervals (Days 1-90, 91-270, 271-450, 451-630, and 631-900.Change of UPDRS-III scores from baseline to each of the five follow-up periods.Change in UPDRS-III scores was significantly greater in PD patients with CRP concentrations ≥0.7 mg/L than in those with CRP concentrations <0.7 mg/L, as determined by a generalized estimation equation model (P = 0.021 for the entire follow-up period and by a generalized regression model (P = 0.030 for the last follow-up interval (Days 631-900. The regression coefficients of baseline CRP for the two periods were 1.41 (95% confidence interval [CI] 0.21-2.61 and 2.62 (95% CI 0.25-4.98, respectively, after adjusting for sex, age, baseline UPDRS-III score, dementia, and incremental L-dopa equivalent dose.Baseline plasma CRP levels were associated with motor deterioration and predicted motor prognosis in patients with PD. These associations were independent of sex, age, PD severity, dementia, and anti-Parkinsonian agents, suggesting that subclinical systemic inflammations could accelerate neurodegeneration in PD.

  8. Influence of energy concentration and source on the utilization of feed protein and NPN in lambs. 3

    International Nuclear Information System (INIS)

    Ulbrich, M.; Geissler, C.; Bassuny, S.M.; Borowiec, F.; Hoffmann, M.

    1989-01-01

    In an N balance experiment with male crossbreeding lambs at an age of 3-4 months four different rations were given differing in energy concentration (high > 700 EFU cattle /kg DM and low cattle /kg DM) and in the energy source (sugar, starch or crude fibre) with crude protein intake being almost equal. The rations contained 2% urea. Microbial protein synthesis in the rumen was assessed according to Roth and Kichgessner (1978) (1), Rys et al. (1975) (2) and Bickel-Baumann and Landis (1986) (3) on the basis of allantoin excretion in urine. The highest ruminal protein synthesis quotas were 868-921 mg protein N per kg LW 0.75 in (2). In (3) 723-766 mg protein N/kg LW 0.75 were synthesized. From the 15 N labelling of the supplemented urea and the excreted allantoin it could be calculated that 26-40% of the microbial protein resulted from the urea-N of the ration. Despite a high crude protein content of the ration of between 16 and 17% in the DM and a relation of NPN: pure protein of 0.95 the utilization of the NPN in the ration was relatively high but slightly lower than the utilization of pure protein. The variants with higher energy concentration showed as a tendency higher allantoin excretion in spite of slightly lower dry matter intake and a slightly higher NPN utilization than the variants with lower energy concentration. (author)

  9. Structural evidence for solvent-stabilisation by aspartic acid as a mechanism for halophilic protein stability in high salt concentrations.

    Science.gov (United States)

    Lenton, Samuel; Walsh, Danielle L; Rhys, Natasha H; Soper, Alan K; Dougan, Lorna

    2016-07-21

    Halophilic organisms have adapted to survive in high salt environments, where mesophilic organisms would perish. One of the biggest challenges faced by halophilic proteins is the ability to maintain both the structure and function at molar concentrations of salt. A distinct adaptation of halophilic proteins, compared to mesophilic homologues, is the abundance of aspartic acid on the protein surface. Mutagenesis and crystallographic studies of halophilic proteins suggest an important role for solvent interactions with the surface aspartic acid residues. This interaction, between the regions of the acidic protein surface and the solvent, is thought to maintain a hydration layer around the protein at molar salt concentrations thereby allowing halophilic proteins to retain their functional state. Here we present neutron diffraction data of the monomeric zwitterionic form of aspartic acid solutions at physiological pH in 0.25 M and 2.5 M concentration of potassium chloride, to mimic mesophilic and halophilic-like environmental conditions. We have used isotopic substitution in combination with empirical potential structure refinement to extract atomic-scale information from the data. Our study provides structural insights that support the hypothesis that carboxyl groups on acidic residues bind water more tightly under high salt conditions, in support of the residue-ion interaction model of halophilic protein stabilisation. Furthermore our data show that in the presence of high salt the self-association between the zwitterionic form of aspartic acid molecules is reduced, suggesting a possible mechanism through which protein aggregation is prevented.

  10. Delipidation of a whey protein concentrate by electroacidification with bipolar membranes.

    Science.gov (United States)

    Shee, Fabrice Lin Teng; Angers, Paul; Bazinet, Laurent

    2007-05-16

    The separation of residual fats from whey protein concentrates (WPC) results in a better nutritional and functional utilization of this product. Bipolar membrane electroacidification (BMEA) technology allows acidification and demineralization of solutions without any salt addition. The principle of BMEA is based on proton formation from water molecule dissociation at the bipolar membrane interface. The objective of this work was to determine the effect of an electroacidification treatment at pH 4.5 on the precipitation of lipids. WPC electroacidification was carried out with or without preliminary demineralization by conventional electrodialysis. The effect of ionic strength on lipid precipitation rates was also evaluated by dilution of the WPC samples. Lipid precipitation levels of 35-39% were obtained using the electroacidification process without a dilution step, while the combination of BMEA and dilution of the WPC resulted in a decrease in lipid content by six-fold from 0.76 to 0.21%.

  11. Effect of ultrasound treatments on functional properties and structure of millet protein concentrate

    DEFF Research Database (Denmark)

    Nazari, Bahman; Mohammadifar, Mohammad Amin; Shojaee-Aliabadi, Saeedeh

    2018-01-01

    In this study, the effect of high power ultrasound (US) probe in varying intensities and times (18.4, 29.58, and 73.95 W/cm2 for 5, 12.5 and 20 min respectively) on functional properties of millet protein concentrate (MPC) was investigated, and also the structural properties of best modified.......37 ± 5.51 ml), but increased upon US treatments at high intensities (749.7 ± 2 ml). In addition, EAI and ES increased after US treatments. One of the best US treatments that can improve the functional properties of MPC was 73.95 W/cm2 for 12.5 min that resulted in reduction of molecular weight and increase...

  12. Evaluation of the protein concentration in enzymes via the determination of sulphur by TXRF

    International Nuclear Information System (INIS)

    Mertens, M.; Rittmeyer, C.; Kolbesen, B.O.

    2000-01-01

    Total reflection x-ray fluorescence spectrometry (TXRF) offers many advantages for the identification of trace elements in biological samples like enzymes, tissues or plants. Without any preliminary treatment elements may be determined with high accuracy especially transition metals like Fe, Ni, Cu, Mo and the alkaline earth metal Ca. A further aspect of the investigation of enzymes is the simple and simultaneous determination of light elements. Especially sulphur is of interest. The element sulphur exists mainly in the two amino, acids methionine and cysteine as well as in iron-sulphur clusters and may be used for an easy and simultaneous calculation of the protein concentration. Hence the quantitative determination of sulphur by TXRF allows a cross-check regarding of the conventional quantitative determination of protein concentration by, for example, the Lowry method. On the basis of three enzymes of different origins and molecular weights the presentation will show the influence of the bio-organic matrix and different buffer media on element determination by TXRF. As is already known the influence of the matrix on the detection of light elements is stronger than on transition metals. It can be discussed whether layer thickness and layer effects of the drying residues (characterization by SEM and thickness profilometer (ALPHA-step)) and / or self absorption effects as well as the excitation are of significance. The results indicate that with enzymes of low molecular weight a reliable determination of sulphur is possible whereas those with higher molecular weights gave poorer results on account of the matrix effects described. (author)

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

    Directory of Open Access Journals (Sweden)

    Chi-Lun Huang

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

  14. C-reactive protein concentration and risk of coronary heart disease, stroke, and mortality: an individual participant meta-analysis

    DEFF Research Database (Denmark)

    Kaptoge, Stephen; Di Angelantonio, Emanuele; Lowe, Gordon

    2010-01-01

    Associations of C-reactive protein (CRP) concentration with risk of major diseases can best be assessed by long-term prospective follow-up of large numbers of people. We assessed the associations of CRP concentration with risk of vascular and non-vascular outcomes under different circumstances....

  15. Dietary modulation of plasma angiopoietin-like protein 4 concentrations in healthy volunteers and in patients with type 2 diabetes

    NARCIS (Netherlands)

    Jonker, Jacqueline T.; Smit, Johannes W. A.; Hammer, Sebastiaan; Snel, Marieke; van der Meer, Rutger W.; Lamb, Hildo J.; Mattijssen, Frits; Mudde, Karin; Jazet, Ingrid M.; Dekkers, Olaf M.; de Roos, Albert; Romijn, Johannes A.; Kersten, Sander; Rensen, Patrick C. N.

    2013-01-01

    Angiopoietin-like protein 4 (ANGPTL4) has been identified as an inhibitor of lipoprotein lipase. Preliminary data suggest that plasma nonesterified fatty acids (NEFAs) raise plasma ANGPTL4 concentrations in humans. The objective was to assess plasma ANGPTL4 concentrations after various nutritional

  16. Dietary modulation of plasma angiopoietin-like protein 4 concentrations in healthy volunteers and in patients with type 2 diabetes

    NARCIS (Netherlands)

    Jonker, J.T.; Smit, J.W.A.; Hammer, S.; Snel, M.; Meer, R.W. van der; Lamb, H.J.; Mattijssen, F.; Mudde, K.; Jazet, I.M.; Dekkers, O.M.; Roos, A. de; Romijn, J.A.; Kersten, S.; Rensen, P.C.

    2013-01-01

    BACKGROUND: Angiopoietin-like protein 4 (ANGPTL4) has been identified as an inhibitor of lipoprotein lipase. Preliminary data suggest that plasma nonesterified fatty acids (NEFAs) raise plasma ANGPTL4 concentrations in humans. OBJECTIVE: The objective was to assess plasma ANGPTL4 concentrations

  17. Dietary modulation of plasma angiopoietin-like protein 4 concentrations in healthy volunteers and in patients with type 2 diabetes

    NARCIS (Netherlands)

    Jonker, J.T.; Smit, J.W.A.; Hammer, S.; Snel, M.; Meer, van der R.; Lamb, H.J.; Mattijssen, F.B.J.; Mudde, C.M.; Jazet, I.M.; Dekkers, O.M.; Roos, de A.; Romijn, J.A.; Kersten, A.H.; Rensen, P.C.N.

    2013-01-01

    Background: Angiopoietin-like protein 4 (ANGPTL4) has been identified as an inhibitor of lipoprotein lipase. Preliminary data suggest that plasma nonesterified fatty acids (NEFAs) raise plasma ANGPTL4 concentrations in humans. Objective: The objective was to assess plasma ANGPTL4 concentrations

  18. Existence of life-time stable proteins in mature rats-Dating of proteins' age by repeated short-term exposure to labeled amino acids throughout age

    DEFF Research Database (Denmark)

    Bechshøft, Cecilie Leidesdorff; Schjerling, Peter; Bornø, Andreas

    2017-01-01

    In vivo turnover rates of proteins covering the processes of protein synthesis and breakdown rates have been measured in many tissues and protein pools using various techniques. Connective tissue and collagen protein turnover is of specific interest since existing results are rather diverging. Th...... living days, indicating very slow turnover. The data support the hypothesis that some proteins synthesized during the early development and growth still exist much later in life of animals and hence has a very slow turnover rate.......In vivo turnover rates of proteins covering the processes of protein synthesis and breakdown rates have been measured in many tissues and protein pools using various techniques. Connective tissue and collagen protein turnover is of specific interest since existing results are rather diverging....... The aim of this study is to investigate whether we can verify the presence of protein pools within the same tissue with very distinct turnover rates over the life-span of rats with special focus on connective tissue. Male and female Lewis rats (n = 35) were injected with five different isotopically...

  19. Selective effects of whey protein concentrate on glutathione levels and apoptosis in rats with mammary tumors.

    Science.gov (United States)

    Cheng, Shih-Hsuan; Tseng, Yang-Ming; Wu, Szu-Hsien; Tsai, Shih-Meng; Tsai, Li-Yu

    2017-09-01

    Glutathione (GSH) plays an important role in antioxidant defense and regulation of apoptosis. GSH deficiency is related to many diseases, including cancer, and increased GSH levels in cancer cells are associated with chemotherapy resistance because of resistance to apoptosis. In this study, we investigated the effects of whey protein concentrate (WPC), a precursor of GSH, in rats with mammary tumors induced by treatment with 7,12-dimethylbenz(a)anthracene (DMBA). DMBA treatment results in cellular changes that mimic the initiation and promotion of carcinogenesis of breast tissue. We aimed to examine the possible preventive effects of diets containing whey protein on DMBA-induced mammary tumors in rats. The results indicate that WPC (0.334 g/kg) supplementation significantly increased the liver GSH levels by 92%, and were accompanied by low Bax/Bcl-2 ratio (from 5 to 3) and cleaved caspase-3/procaspase-3 ratio (from 2.4 to 1.2) in DMBA-treated rats. Furthermore, tumor GSH levels were decreased by 47% in WPC-supplemented rats, which resulted in increased Bax/Bcl-2 ratio (from 0.9 to 2) and cleaved caspase-3/procaspase-3 ratio (from 1.1 to 2.7). In conclusion, supplementation with WPC could selectively deplete tumor GSH levels and, therefore, WPC supplementation might be a promising strategy to overcome treatment resistance in cancer therapy. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Effects of incorporation of whey protein concentrate on physicochemical, texture, and microbial evaluation of developed cookies

    Directory of Open Access Journals (Sweden)

    Safa Hamid Wani

    2015-12-01

    Full Text Available Whey Protein concentrate (WPC was incorporated into cookies at different levels (0, 2, 4, and 6%. Cookies were analyzed for physicochemical, color, textural, microbial, and sensory attributes. Physicochemical analysis revealed that 6% WPC supplemented cookies shows maximum protein content (13.22%, moisture content (11.33%, fat content (23.08%, and ash content (2.02% as compared to control. However, control sample shows significantly different (p ≤ 0.05 value for crude fiber and carbohydrate content. Maximum thickness (9.63 mm, diameter (44.06 mm, and weight (9.10 g were found for control and these decreased significantly (p ≤ 0.05 with increase in WPC supplementation level in cookies. Cookie supplemented with 4% WPC showed maximum overall acceptability (4.76. Texture analysis revealed that 6% WPC supplemented cookie shows maximum cutting force (55.3 N. Lightness (L* value of cookies decreased from 67.32 to 57.94. Where as a* and b* value increased from 0.37 to 3.57 and 25.35 to 27.54, respectively. The total plate count of cookie samples was under acceptable limits.

  1. A fluorescence anisotropy method for measuring protein concentration in complex cell culture media.

    Science.gov (United States)

    Groza, Radu Constantin; Calvet, Amandine; Ryder, Alan G

    2014-04-22

    The rapid, quantitative analysis of the complex cell culture media used in biopharmaceutical manufacturing is of critical importance. Requirements for cell culture media composition profiling, or changes in specific analyte concentrations (e.g. amino acids in the media or product protein in the bioprocess broth) often necessitate the use of complicated analytical methods and extensive sample handling. Rapid spectroscopic methods like multi-dimensional fluorescence (MDF) spectroscopy have been successfully applied for the routine determination of compositional changes in cell culture media and bioprocess broths. Quantifying macromolecules in cell culture media is a specific challenge as there is a need to implement measurements rapidly on the prepared media. However, the use of standard fluorescence spectroscopy is complicated by the emission overlap from many media components. Here, we demonstrate how combining anisotropy measurements with standard total synchronous fluorescence spectroscopy (TSFS) provides a rapid, accurate quantitation method for cell culture media. Anisotropy provides emission resolution between large and small fluorophores while TSFS provides a robust measurement space. Model cell culture media was prepared using yeastolate (2.5 mg mL(-1)) spiked with bovine serum albumin (0 to 5 mg mL(-1)). Using this method, protein emission is clearly discriminated from background yeastolate emission, allowing for accurate bovine serum albumin (BSA) quantification over a 0.1 to 4.0 mg mL(-1) range with a limit of detection (LOD) of 13.8 μg mL(-1). Copyright © 2014. Published by Elsevier B.V.

  2. Whey protein concentrate supplementation protects rat brain against aging-induced oxidative stress and neurodegeneration.

    Science.gov (United States)

    Garg, Geetika; Singh, Sandeep; Singh, Abhishek Kumar; Rizvi, Syed Ibrahim

    2018-05-01

    Whey protein concentrate (WPC) is a rich source of sulfur-containing amino acids and is consumed as a functional food, incorporating a wide range of nutritional attributes. The purpose of this study is to evaluate the neuroprotective effect of WPC on rat brain during aging. Young (4 months) and old (24 months) male Wistar rats were supplemented with WPC (300 mg/kg body weight) for 28 days. Biomarkers of oxidative stress and antioxidant capacity in terms of ferric reducing antioxidant potential (FRAP), lipid hydroperoxide (LHP), total thiol (T-SH), protein carbonyl (PC), reactive oxygen species (ROS), nitric oxide (NO), and acetylcholinesterase (AChE) activity were measured in brain of control and experimental (WPC supplemented) groups. In addition, gene expression and histopathological studies were also performed. The results indicate that WPC augmented the level of FRAP, T-SH, and AChE in old rats as compared with the old control. Furthermore, WPC-treated groups exhibited significant reduction in LHP, PC, ROS, and NO levels in aged rats. WPC supplementation also downregulated the expression of inflammatory markers (tumor necrosis factor alpha, interleukin (IL)-1β, IL-6), and upregulated the expression of marker genes associated with autophagy (Atg3, Beclin-1, LC3B) and neurodegeneration (neuron specific enolase, Synapsin-I, MBP-2). The findings suggested WPC to be a potential functional nutritional food supplement that prevents the progression of age-related oxidative damage in Wistar rats.

  3. Distribution of radionuclides in leaf-stem biomass of lupine and clover under production of protein concentrates

    International Nuclear Information System (INIS)

    Novikov, Yu.F.; Lobach, G.A.; Buzenko, T.A.; Zaretskaya, T.P.

    1993-01-01

    The basic regularities of radionuclide distribution between the obtained products have been studied using the fractionation of lupine and clover phytomass as an example. The content of radionuclides in protein concentrates has been shown to be strongly related to the crop species. A scheme and a regime of the fractionation of leaf-stem lupine biomass contaminated with cesium radioisotopes and strontium-90 which ensured the minimizing of their residual content in protein-vitaminic and protein concentrates have been selected with due accout of experimental data

  4. Stable transformation of sunflower (Helianthus annuus L.) using a non-meristematic regeneration protocol and green fluorescent protein as a vital marker.

    Science.gov (United States)

    Müller, A; Iser, M; Hess, D

    2001-10-01

    Stable transformation of sunflower was achieved using a non-meristematic hypocotyl explant regeneration protocol of public inbred HA300B. Uniformly transformed shoots were obtained after co-cultivation with Agrobacterium tumefaciens carrying a gfp (green fluorescent protein) gene containing an intron that blocks expression of gfp in Agrobacterium. Easily detectable, bright green fluorescence of transformed tissues was used to establish an optimal regeneration and transformation procedure. By Southern blot analysis, integration of the gfp and nptll genes was confirmed. Stable transformation efficiency was 0.1%. From 68 T1 plants analyzed, 17 showed transmission of transgene DNA and 15 of them contained the intact gfp gene. Expression of gfp was detected in 10 T1 plants carrying the intact gfp gene using a fluorimetric assay or western blot analysis. Expression of the nptll gene was confirmed in 13 T1 plants. The transformation system enables the rapid transfer of agronomically important genes.

  5. Usefulness of estimation of blood procalcitonin concentration versus C-reactive protein concentration and white blood cell count for therapeutic monitoring of sepsis in neonates

    Directory of Open Access Journals (Sweden)

    Agnieszka Kordek

    2014-12-01

    Full Text Available Aim: This study was intended to assess the clinical usefulness of blood procalcitonin (PCT concentrations for the diagnosis and therapeutic monitoring of nosocomial neonatal sepsis.Material/Methods: The enrolment criterion was sepsis clinically manifesting after three days of life. PCT concentrations were measured in venous blood from 52 infected and 88 uninfected neonates. The results were interpreted against C-reactive protein (CRP concentrations and white blood cell counts (WBC.Results: Differences between the two groups in PCT and CRP concentrations were highly significant. No significant differences between the groups were noted for WBC. The threshold value on the receiver operator characteristic curve was 2.06 ng/mL for PCT (SE 75%; SP 80.68%; PPV 62.22%; NPV 88.75%; AUC 0.805, 5.0 mg/L for CRP (SE 67.44%; SP 73.68%; PPV 42.02%; NPV 88.89%; AUC 0.801, and 11.9 x109/L for WBC (SE 51.16%; SP 50.68%; PPV 23.16%; NPV 78.13%; AUC 0.484. Procalcitonin concentrations decreased 24 hours after initiation of antibiotic therapy and reverted to the control level after 5-7 days. C-reactive protein concentrations began to decline after two days of antibiotic therapy but were still higher than in the control group after 5-7 days of treatment. No significant changes in WBC during the treatment were observed.Conclusions: Procalcitonin concentrations in blood appear to be of use for the diagnosis and therapeutic monitoring of nosocomial infections in neonates as this parameter demonstrates greater sensitivity and specificity than C-reactive protein. White blood cell counts appear to be of little diagnostic value in the early phase of infection or for therapeutic monitoring.

  6. Bone marrow concentrate and platelet-rich plasma differ in cell distribution and interleukin 1 receptor antagonist protein concentration.

    Science.gov (United States)

    Cassano, Jennifer M; Kennedy, John G; Ross, Keir A; Fraser, Ethan J; Goodale, Margaret B; Fortier, Lisa A

    2018-01-01

    Bone marrow concentrate (BMC) and platelet-rich plasma (PRP) are used extensively in regenerative medicine. The aim of this study was to determine differences in the cellular composition and cytokine concentrations of BMC and PRP and to compare two commercial BMC systems in the same patient cohort. Patients (29) undergoing orthopaedic surgery were enrolled. Bone marrow aspirate (BMA) was processed to generate BMC from two commercial systems (BMC-A and BMC-B). Blood was obtained to make PRP utilizing the same system as BMC-A. Bone marrow-derived samples were cultured to measure colony-forming units, and flow cytometry was performed to assess mesenchymal stem cell (MSC) markers. Cellular concentrations were assessed for all samples. Catabolic cytokines and growth factors important for cartilage repair were measured using multiplex ELISA. Colony-forming units were increased in both BMCs compared to BMA (p BMC-A and PRP, but there were differences in leucocyte concentrations. TGF-β1 and PDGF were not different between BMC-A and PRP. IL-1ra concentrations were greater (p = 0.0018) in BMC-A samples (13,432 pg/mL) than in PRP (588 pg/mL). The IL-1ra/IL-1β ratio in all BMC samples was above the value reported to inhibit IL-1β. The bioactive factors examined in this study have differing clinical effects on musculoskeletal tissue. Differences in the cellular and cytokine composition between PRP and BMC and between BMC systems should be taken into consideration by the clinician when choosing a biologic for therapeutic application. Clinical, Level II.

  7. Total protein concentration and diagnostic test results for gray wolf (Canis lupus) serum using Nobuto filter paper strips

    Science.gov (United States)

    Jara, Rocio F.; Sepúlveda, Carolina; Ip, Hon S.; Samuel, Michael D.

    2015-01-01

    Nobuto filter paper strips are widely used for storing blood-serum samples, but the recovery of proteins from these strips following rehydration is unknown. Poor recovery of proteins could reduce the concentration of antibodies and antigens and reduce the sensitivity of diagnostic assays. We compared the protein concentration, and its association with test sensitivity, of eluted Nobuto strip samples with paired sera. We collected and froze serum from five gray wolves (Canis lupus) for 8 mo. When thawed, we used a spectrophotometer (absorbance 280 nm) to determine the serum protein concentration for paired sera and Nobuto eluates for each animal in 2-fold serial dilutions. Total protein concentration was similar for both sample storage methods (Nobuto eluates and control sera), except for the undiluted samples in which Nobuto eluates had higher total protein concentrations. Both sample storage methods appear to produce similar results using the SNAP® 4Dx® Test to detect antibodies against pathogens causing Lyme disease, anaplasmosis, and ehrlichiosis as well as antigen for canine heartworm disease.

  8. Protein brownian rotation at the glass transition temperature of a freeze-concentrated buffer probed by superparamagnetic nanoparticles.

    Science.gov (United States)

    Eloi, J-C; Okuda, M; Jones, S E Ward; Schwarzacher, W

    2013-06-18

    For applications from food science to the freeze-thawing of proteins it is important to understand the often complex freezing behavior of solutions of biomolecules. Here we use a magnetic method to monitor the Brownian rotation of a quasi-spherical cage-shaped protein, apoferritin, approaching the glass transition Tg in a freeze-concentrated buffer (Tris-HCl). The protein incorporates a synthetic magnetic nanoparticle (Co-doped Fe3O4 (magnetite)). We use the magnetic signal from the nanoparticles to monitor the protein orientation. As T decreases toward Tg of the buffer solution the protein's rotational relaxation time increases exponentially, taking values in the range from a few seconds up to thousands of seconds, i.e., orders of magnitude greater than usually accessed, e.g., by NMR. The longest relaxation times measured correspond to estimated viscosities >2 MPa s. As well as being a means to study low-temperature, high-viscosity environments, our method provides evidence that, for the cooling protocol used, the following applies: 1), the concentration of the freeze-concentrated buffer at Tg is independent of its initial concentration; 2), little protein adsorption takes place at the interface between ice and buffer; and 3), the protein is free to rotate even at temperatures as low as 207 K. Copyright © 2013 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  9. Increased Milk Protein Concentration in a Rehydration Drink Enhances Fluid Retention Caused by Water Reabsorption in Rats.

    Science.gov (United States)

    Ito, Kentaro; Saito, Yuri; Ashida, Kinya; Yamaji, Taketo; Itoh, Hiroyuki; Oda, Munehiro

    2015-01-01

    A fluid-retention effect is required for beverages that are designed to prevent dehydration. That is, fluid absorbed from the intestines should not be excreted quickly; long-term retention is desirable. Here, we focused on the effect of milk protein on fluid retention, and propose a new effective oral rehydration method that can be used daily for preventing dehydration. We first evaluated the effects of different concentrations of milk protein on fluid retention by measuring the urinary volumes of rats fed fluid containing milk protein at concentrations of 1, 5, and 10%. We next compared the fluid-retention effect of milk protein-enriched drink (MPD) with those of distilled water (DW) and a sports drink (SD) by the same method. Third, to investigate the mechanism of fluid retention, we measured plasma insulin changes in rats after ingesting these three drinks. We found that the addition of milk protein at 5 or 10% reduced urinary volume in a dose-dependent manner. Ingestion of the MPD containing 4.6% milk protein resulted in lower urinary volumes than DW and SD. MPD also showed a higher water reabsorption rate in the kidneys and higher concentrations of plasma insulin than DW and SD. These results suggest that increasing milk protein concentration in a beverage enhances fluid retention, which may allow the possibility to develop rehydration beverages that are more effective than SDs. In addition, insulin-modifying renal water reabsorption may contribute to the fluid-retention effect of MPD.

  10. The effect of microfiltration on color, flavor, and functionality of 80% whey protein concentrate.

    Science.gov (United States)

    Qiu, Y; Smith, T J; Foegeding, E A; Drake, M A

    2015-09-01

    The residual annatto colorant in fluid Cheddar cheese whey is bleached to provide a neutral-colored final product. Currently, hydrogen peroxide (HP) and benzoyl peroxide are used for bleaching liquid whey. However, previous studies have shown that chemical bleaching causes off-flavor formation, mainly due to lipid oxidation and protein degradation. The objective of this study was to evaluate the efficacy of microfiltration (MF) on norbixin removal and to compare flavor and functionality of 80% whey protein concentrate (WPC80) from MF whey to WPC80 from whey bleached with HP or lactoperoxidase (LP). Cheddar cheese whey was manufactured from colored, pasteurized milk. The fluid whey was pasteurized and fat separated. Liquid whey was subjected to 4 different treatments: control (no bleaching; 50°C, 1 h), HP (250 mg of HP/kg; 50°C, 1 h), and LP (20 mg of HP/kg; 50°C, 1 h), or MF (microfiltration; 50°C, 1 h). The treated whey was then ultrafiltered, diafiltered, and spray-dried to 80% concentrate. The entire experiment was replicated 3 times. Proximate analyses, color, functionality, descriptive sensory and instrumental volatile analysis were conducted on WPC80. The MF and HP- and LP-bleached WPC80 displayed a 39.5, 40.9, and 92.8% norbixin decrease, respectively. The HP and LP WPC80 had higher cardboard flavors and distinct cabbage flavor compared with the unbleached and MF WPC80. Volatile compound results were consistent with sensory results. The HP and LP WPC80 were higher in lipid oxidation compounds (especially heptanal, hexanal, pentanal, 1-hexen-3-one, 2-pentylfuran, and octanal) compared with unbleached and MF WPC80. All WPC80 had >85% solubility across the pH range of 3 to 7. The microstructure of MF gels determined by confocal laser scanning showed an increased protein particle size in the gel network. MF WPC80 also had larger storage modulus values, indicating higher gel firmness. Based on bleaching efficacy comparable to chemical bleaching with HP

  11. Molecular environment of stable iodine and radioiodine (129I) in natural organic matter: Evidence inferred from NMR and binding experiments at environmentally relevant concentrations

    Science.gov (United States)

    Xu, Chen; Zhong, Junyan; Hatcher, Patrick G.; Zhang, Saijin; Li, Hsiu-Ping; Ho, Yi-Fang; Schwehr, Kathleen A.; Kaplan, Daniel I.; Roberts, Kimberly A.; Brinkmeyer, Robin; Yeager, Chris M.; Santschi, Peter H.

    2012-11-01

    129I is a major by-product of nuclear fission and had become one of the major radiation risk drivers at Department of Energy (DOE) sites. 129I is present at elevated levels in the surface soils of the Savannah River Site (SRS) F-Area and was found to be bound predominantly to soil organic matter (SOM). Naturally bound 127I and 129I to sequentially extracted humic acids (HAs), fulvic acids (FAs) and a water extractable colloid (WEC) were measured in a 129I-contaminated wetland surface soil located on the SRS. WEC is a predominantly colloidal organic fraction obtained from soil re-suspension experiments to mimic the fraction that may be released during groundwater exfiltration, storm water or surface runoff events. For the first time, NMR techniques were applied to infer the molecular environment of naturally occurring stable iodine and radioiodine binding to SOM. Iodine uptake partitioning coefficients (Kd) by these SOM samples at ambient iodine concentrations were also measured and related to quantitative structural analyses by 13C DPMAS NMR and solution state 1H NMR on the eight humic acid fractions. By assessing the molecular environment of iodine, it was found that it was closely associated with the aromatic regions containing esterified products of phenolic and formic acids or other aliphatic carboxylic acids, amide functionalities, quinone-like structures activated by electron-donating groups (e.g., NH2), or a hemicellulose-lignin-like complex with phenyl-glycosidic linkages. However, FAs and WEC contained much greater concentrations of 127I or 129I than HAs. The contrasting radioiodine contents among the three different types of SOM (HAs, FAs and WEC) suggest that the iodine binding environment cannot be explained solely by the difference in the amount of their reactive binding sites. Instead, indirect evidence indicates that the macro-molecular conformation, such as the hydrophobic aliphatic periphery hindering the active aromatic cores and the hydrophilic

  12. Stability of buffer-free freeze-dried formulations: A feasibility study of a monoclonal antibody at high protein concentrations.

    Science.gov (United States)

    Garidel, Patrick; Pevestorf, Benjamin; Bahrenburg, Sven

    2015-11-01

    We studied the stability of freeze-dried therapeutic protein formulations over a range of initial concentrations (from 40 to 160 mg/mL) and employed a variety of formulation strategies (including buffer-free freeze dried formulations, or BF-FDF). Highly concentrated, buffer-free liquid formulations of therapeutic monoclonal antibodies (mAbs) have been shown to be a viable alternative to conventionally buffered preparations. We considered whether it is feasible to use the buffer-free strategy in freeze-dried formulations, as an answer to some of the known drawbacks of conventional buffers. We therefore conducted an accelerated stability study (24 weeks at 40 °C) to assess the feasibility of stabilizing freeze-dried formulations without "classical" buffer components. Factors monitored included pH stability, protein integrity, and protein aggregation. Because the protein solutions are inherently self-buffering, and the system's buffer capacity scales with protein concentration, we included highly concentrated buffer-free freeze-dried formulations in the study. The tested formulations ranged from "fully formulated" (containing both conventional buffer and disaccharide stabilizers) to "buffer-free" (including formulations with only disaccharide lyoprotectant stabilizers) to "excipient-free" (with neither added buffers nor stabilizers). We evaluated the impacts of varying concentrations, buffering schemes, pHs, and lyoprotectant additives. At the end of 24 weeks, no change in pH was observed in any of the buffer-free formulations. Unbuffered formulations were found to have shorter reconstitution times and lower opalescence than buffered formulations. Protein stability was assessed by visual inspection, sub-visible particle analysis, protein monomer content, charge variants analysis, and hydrophobic interaction chromatography. All of these measures found the stability of buffer-free formulations that included a disaccharide stabilizer comparable to buffer

  13. Susceptibility to viral infection is enhanced by stable expression of 3A or 3AB proteins from foot-and-mouth disease virus

    International Nuclear Information System (INIS)

    Rosas, Maria F.; Vieira, Yuri A.; Postigo, Raul; Martin-Acebes, Miguel A.; Armas-Portela, Rosario; Martinez-Salas, Encarnacion; Sobrino, Francisco

    2008-01-01

    The foot-and-mouth disease virus (FMDV) 3A protein is involved in virulence and host range. A distinguishing feature of FMDV 3B among picornaviruses is that three non-identical copies are encoded in the viral RNA and required for optimal replication in cell culture. Here, we have studied the involvement of the 3AB region on viral infection using constitutive and transient expression systems. BHK-21 stably transformed clones expressed low levels of FMDV 3A or 3A(B) proteins in the cell cytoplasm. Transformed cells stably expressing these proteins did not exhibit inner cellular rearrangements detectable by electron microscope analysis. Upon FMDV infection, clones expressing either 3A alone or 3A(B) proteins showed a significant increase in the percentage of infected cells, the number of plaque forming units and the virus yield. The 3A-enhancing effect was specific for FMDV as no increase in viral multiplication was observed in transformed clones infected with another picornavirus, encephalomyocarditis virus, or the negative-strand RNA virus vesicular stomatitis virus. A potential role of 3A protein in viral RNA translation was discarded by the lack of effect on FMDV IRES-dependent translation. Increased viral susceptibility was not caused by a released factor; neither the supernatant of transformed clones nor the addition of purified 3A protein to the infection medium was responsible for this effect. Unlike stable expression, high levels of 3A or 3A(B) protein transient expression led to unspecific inhibition of viral infection. Therefore, the effect observed on viral yield, which inversely correlated with the intracellular levels of 3A protein, suggests a transacting role operating on the FMDV multiplication cycle

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

    Science.gov (United States)

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

    2014-05-01

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

  15. Proposal for a coordination research programme (CRP) of the International Atomic Energy Agency (IAEA) on stable isotope tracer techniques for studies on protein-energy interactions

    International Nuclear Information System (INIS)

    Shetty, P.; James, W.P.T.

    1993-01-01

    This Report provides a rationale and justification for the initiation of a Coordinated Research programme to support studies using stable isotopic tracer techniques to address priority areas of human protein-energy interactions with special emphasis on the problems of human nutrition in developing countries. The Report suggests a modus for establishing such a practically oriented Coordinated Research Programme under the aegis of the International Atomic Energy Agency with concrete suggestions for its organization and the identification of probable participants in such a programme. The likely sources of additional funding to sustain such an activity viable for a period of 4 to 5 years are also indicated. 8 refs

  16. System in biology leading to cell pathology: stable protein-protein interactions after covalent modifications by small molecules or in transgenic cells.

    Science.gov (United States)

    Malina, Halina Z

    2011-01-19

    The physiological processes in the cell are regulated by reversible, electrostatic protein-protein interactions. Apoptosis is such a regulated process, which is critically important in tissue homeostasis and development and leads to complete disintegration of the cell. Pathological apoptosis, a process similar to apoptosis, is associated with aging and infection. The current study shows that pathological apoptosis is a process caused by the covalent interactions between the signaling proteins, and a characteristic of this pathological network is the covalent binding of calmodulin to regulatory sequences. Small molecules able to bind covalently to the amino group of lysine, histidine, arginine, or glutamine modify the regulatory sequences of the proteins. The present study analyzed the interaction of calmodulin with the BH3 sequence of Bax, and the calmodulin-binding sequence of myristoylated alanine-rich C-kinase substrate in the presence of xanthurenic acid in primary retinal epithelium cell cultures and murine epithelial fibroblast cell lines transformed with SV40 (wild type [WT], Bid knockout [Bid-/-], and Bax-/-/Bak-/- double knockout [DKO]). Cell death was observed to be associated with the covalent binding of calmodulin, in parallel, to the regulatory sequences of proteins. Xanthurenic acid is known to activate caspase-3 in primary cell cultures, and the results showed that this activation is also observed in WT and Bid-/- cells, but not in DKO cells. However, DKO cells were not protected against death, but high rates of cell death occurred by detachment. The results showed that small molecules modify the basic amino acids in the regulatory sequences of proteins leading to covalent interactions between the modified sequences (e.g., calmodulin to calmodulin-binding sites). The formation of these polymers (aggregates) leads to an unregulated and, consequently, pathological protein network. The results suggest a mechanism for the involvement of small molecules

  17. Protein remains stable at unusually high temperatures when solvated in aqueous mixtures of amino acid based ionic liquids

    DEFF Research Database (Denmark)

    Chevrot, Guillaume; Fileti, Eudes Eterno; Chaban, Vitaly V.

    2016-01-01

    Using molecular dynamics simulations, we investigated the thermal stability and real-time denaturation of a model mini-protein in four solvents: (1) water, (2) 1-ethyl-3-methylimidazolium alaninate [EMIM][ALA] (5 mol% in water), (3) methioninate [EMIM][MET] (5 mol% in water), and (4) tryptophanat...... (AAILs) than in water. This thermal stability was correlated with the thermodynamics and shear viscosity of the AAIL-containing mixtures. These results suggest that AAILs are generally favorable for protein conservation. [Figure not available: see fulltext.]...

  18. Serum levels of C-reactive protein in patients with stable coronary artery disease: JUPITER in perspective.

    Science.gov (United States)

    Saely, Christoph H; Rein, Philipp; Vonbank, Alexander; Drexel, Heinz

    2010-10-29

    The JUPITER trial has recently demonstrated an outstanding reduction of cardiovascular events by 20 mg rosuvastatin/day in subjects with high CRP who were apparently healthy at baseline. However, absence of atherosclerosis in JUPITER was based on the subjects' history and not proven objectively. To put the results of JUPITER in perspective, we evaluated serum CRP in a consecutive series of 703 statin-naïve Caucasian patients with angiographically proven stable CAD. From these stable CAD patients, only 69.2% met the ≥2.0 mg/l serum CRP inclusion criterion of the JUPITER trial. Median CRP [interquartile range] in our CAD patients was 3.3 [1.6-6.6] mg/l, which was significantly (pJUPITER (4.2 mg/l). Our results point to considerable subclinical atherosclerosis in the patients studied in JUPITER. The impressive results of that trial may not be generalizable to healthy populations all over the world. Copyright © 2009 Elsevier Ireland Ltd. All rights reserved.

  19. Fish Protein Concentrate Fortification Siam Patin on Amplang Snack Products and Mi Sago Instant Product as a Leading Regional Riau

    Directory of Open Access Journals (Sweden)

    Dewita Buchari

    2014-11-01

    Full Text Available To enhance fish consumption in the community especially children, fortification on processed fish product is conducted. The processed fish products are developed to fill the requirements as the fish based food products that own characterizations such as ready to eat, easy to carry, and less time to cook. Amplang snacks and instant sagoo noodles are defined as the products that fills the requirements. The research was aimed to process catfish into fish protein concentrate to become amplang snack and instant sagoo noodles. These products were designed as the effort to develop the local priority products in Riau by using diversification and fortification methods. Experimental method with fortification treatments on Fish Protein Concentrate (FPC extract from Catfish that generate products of amplang snacks and instant sagoo noodles and fish tofu were carried out. The fortified products were examined by organoleptics test that involved panelists. The results showed that the proximate analysis on fortified Catfish Protein Concentrate products were presented as following :1. water contents of 3,13 %, ash of 2,85 %, protein content of 16,13 % and fat content of 18, 66 % for ampang snacks; and 2. water contents of 11,77 %, ash of 1,30 %, protein content of 12,35 % and fat content of 1,86 % for instant sagoo nodles. All fortified FPC products filled the Indonesian Nasional Standard (SNI.Keywords: Fortification, Catfish, and Fish Protein Concentrate

  20. Quantitative Proteomics Analysis of VEGF-Responsive Endothelial Protein S-Nitrosylation Using Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) and LC-MS/MS.

    Science.gov (United States)

    Zhang, Hong-Hai; Lechuga, Thomas J; Chen, Yuezhou; Yang, Yingying; Huang, Lan; Chen, Dong-Bao

    2016-05-01

    Adduction of a nitric oxide moiety (NO•) to cysteine(s), termed S-nitrosylation (SNO), is a novel mechanism for NO to regulate protein function directly. However, the endothelial SNO-protein network that is affected by endogenous and exogenous NO is obscure. This study was designed to develop a quantitative proteomics approach using stable isotope labeling by amino acids in cell culture for comparing vascular endothelial growth factor (VEGFA)- and NO donor-responsive endothelial nitroso-proteomes. Primary placental endothelial cells were labeled with "light" (L-(12)C6 (14)N4-Arg and L-(12)C6 (14)N2-Lys) or "heavy" (L-(13)C6 (15)N4-Arg and L-(13)C6 (15)N2-Lys) amino acids. The light cells were treated with an NO donor nitrosoglutathione (GSNO, 1 mM) or VEGFA (10 ng/ml) for 30 min, while the heavy cells received vehicle as control. Equal amounts of cellular proteins from the light (GSNO or VEGFA treated) and heavy cells were mixed for labeling SNO-proteins by the biotin switch technique and then trypsin digested. Biotinylated SNO-peptides were purified for identifying SNO-proteins by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Ratios of light to heavy SNO-peptides were calculated for determining the changes of the VEGFA- and GSNO-responsive endothelial nitroso-proteomes. A total of 387 light/heavy pairs of SNO-peptides were identified, corresponding to 213 SNO-proteins that include 125 common and 27 VEGFA- and 61 GSNO-responsive SNO-proteins. The specific SNO-cysteine(s) in each SNO-protein were simultaneously identified. Pathway analysis revealed that SNO-proteins are involved in various endothelial functions, including proliferation, motility, metabolism, and protein synthesis. We collectively conclude that endogenous NO on VEGFA stimulation and exogenous NO from GSNO affect common and different SNO-protein networks, implicating SNO as a critical mechanism for VEGFA stimulation of angiogenesis. © 2016 by the Society for the Study of Reproduction

  1. Albumen foam stability and s-ovalbumin contents in eggs coated with whey protein concentrate

    Directory of Open Access Journals (Sweden)

    ACC Alleoni

    2004-06-01

    Full Text Available Food products such as breads, cakes, crackers, meringues, ice creams and several bakery items depend on air incorporation to maintain their texture and structure during or after processing. Proteins are utilized in the food industry since they improve texture attributes through their ability to encapsulate and retain air. The objectives of this work were to quantify s-ovalbumin contents in albumen and to determine alterations in egg white foam stability in fresh eggs, and in eggs coated and non-coated with a whey protein-based concentrate film (WPC, stored at 25°C for 28 days. The volume of drained liquid was higher in non-coated eggs than in coated eggs stored at 25°C at all storage periods. The difference on the third day of storage was in the order of 59% between coated and non-coated eggs, while on the twenty-eighth day it was 202%. During the storage period, an increase in pH and drainage volume was observed for non-coated eggs. After three days, the non-coated eggs showed a s-ovalbumin content 33% higher than coated eggs; this increase jumped to 205% at 28 days of storage. There was a positive correlation between s-ovalbumin content and the volume of drained liquid for coated and non-coated eggs; in other words, when the s-ovalbumin content increased, there was an increase in the volume of drained liquid and a decrease in foam stability. WPC coating maintain egg quality, since it is an effective barrier against the loss of CO2, avoiding changes in the pH of egg white.

  2. A novel approach for the detection of potentially hazardous pepsin stable hazelnut proteins as contaminants in chocolate-based food

    NARCIS (Netherlands)

    Akkerdaas, Jaap H.; Wensing, Marjolein; Knulst, André C.; Stephan, Oliver; Hefle, Susan L.; Aalberse, Rob C.; van Ree, Ronald

    2004-01-01

    Contamination of food products with pepsin resistant allergens is generally believed to be a serious threat to patients with severe food allergy. A sandwich type enzyme-linked immunosorbent assay (ELISA) was developed to measure pepsin resistant hazelnut protein in food products. Capturing and

  3. C-reactive protein and angiographic characteristics of stable and unstable coronary artery disease : Data from the prospective PREVEND cohort

    NARCIS (Netherlands)

    Geluk, C.A.; Post, W.J.; Hillege, H.L.; Tio, R.A.; Tijssen, J.G.; van Dijk, R.B.; Dijk, Willem; Bakker, S.J.; de Jong, P.E.; van Gilst, W.H.; Zijlstra, F.

    Aims: High sensitive-C-reactive protein (hs-CRP) is associated with coronary risk, which may be explained by an association with (unstable) coronary artery disease (CAD). Until now, histopathological and angiographic studies have failed to consistently demonstrate a strong relationship. However,

  4. Influence of energy concentration and source on the utilization of feed protein and NPN in lambs. 3. Allantoin excretion and microbial protein synthesis

    Energy Technology Data Exchange (ETDEWEB)

    Ulbrich, M; Geissler, C; Bassuny, S M; Borowiec, F; Hoffmann, M

    1989-06-01

    In an N balance experiment with male crossbreeding lambs at an age of 3-4 months four different rations were given differing in energy concentration (high > 700 EFU/sub cattle//kg DM and low < 650 EFU/sub cattle//kg DM) and in the energy source (sugar, starch or crude fibre) with crude protein intake being almost equal. The rations contained 2% urea. Microbial protein synthesis in the rumen was assessed according to Roth and Kichgessner (1978) (1), Rys et al. (1975) (2) and Bickel-Baumann and Landis (1986) (3) on the basis of allantoin excretion in urine. The highest ruminal protein synthesis quotas were 868-921 mg protein N per kg LW/sup 0.75/ in (2). In (3) 723-766 mg protein N/kg LW /sup 0.75/ were synthesized. From the /sup 15/N labelling of the supplemented urea and the excreted allantoin it could be calculated that 26-40% of the microbial protein resulted from the urea-N of the ration. Despite a high crude protein content of the ration of between 16 and 17% in the DM and a relation of NPN: pure protein of 0.95 the utilization of the NPN in the ration was relatively high but slightly lower than the utilization of pure protein. The variants with higher energy concentration showed as a tendency higher allantoin excretion in spite of slightly lower dry matter intake and a slightly higher NPN utilization than the variants with lower energy concentration. (author).

  5. Osmolyte Effects on Monoclonal Antibody Stability and Concentration-Dependent Protein Interactions with Water and Common Osmolytes.

    Science.gov (United States)

    Barnett, Gregory V; Razinkov, Vladimir I; Kerwin, Bruce A; Blake, Steven; Qi, Wei; Curtis, Robin A; Roberts, Christopher J

    2016-04-07

    Preferential interactions of proteins with water and osmolytes play a major role in controlling the thermodynamics of protein solutions. While changes in protein stability and shifts in phase behavior are often reported with the addition of osmolytes, the underlying protein interactions with water and/or osmolytes are typically inferred rather than measured directly. In this work, Kirkwood-Buff integrals for protein-water interactions (G12) and protein-osmolyte interactions (G23) were determined as a function of osmolyte concentration from density measurements of antistreptavidin immunoglobulin gamma-1 (AS-IgG1) in ternary aqueous solutions for a set of common neutral osmolytes: sucrose, trehalose, sorbitol, and poly(ethylene glycol) (PEG). For sucrose and PEG solutions, both protein-water and protein-osmolyte interactions depend strongly on osmolyte concentrations (c3). Strikingly, both osmolytes change from being preferentially excluded to preferentially accumulated with increasing c3. In contrast, sorbitol and trehalose solutions do not show large enough preferential interactions to be detected by densimetry. G12 and G23 values are used to estimate the transfer free energy for native AS-IgG1 (Δμ2N) and compared with existing models. AS-IgG1 unfolding via calorimetry shows a linear increase in midpoint temperatures as a function of trehalose, sucrose, and sorbitol concentrations, but the opposite behavior for PEG. Together, the results highlight limitations of existing models and common assumptions regarding the mechanisms of protein stabilization by osmolytes. Finally, PEG preferential interactions destabilize the Fab regions of AS-IgG1 more so than the CH2 or CH3 domains, illustrating preferential interactions can be specific to different protein domains.

  6. Metabolisable protein supply to lactating dairy cows increased with increasing dry matter concentration in grass-clover silage

    DEFF Research Database (Denmark)

    Johansen, Marianne; Hellwing, Anne Louise Frydendahl; Lund, Peter

    2017-01-01

    The aim of this experiment was to study the effect of increased dry matter (DM) concentration in grass-clover silage, obtained by extending the pre-wilting period before ensiling, on the amount of metabolisable protein (MP) supplied to lactating dairy cows. Spring growth and first regrowth of grass...... and faeces, respectively, were collected over 94 h to cover the diurnal variation, pooled, and subsequently analysed. Rumen fluid was collected in same sampling procedure. To estimate the duodenal flow of microbial protein, microbes were isolated from the rumen and analysed for amino acids (AA) and purines...... flow of AA. The higher duodenal flow of AA derived from a lower rumen degradation of feed protein and a tendency towards a higher microbial synthesis in the rumen. Fibre digestibility and CH4 production were not affected by silage DM concentration. In conclusion, MP concentration in grass-clover silage...

  7. Physicochemical and functional properties of protein concentrate from by-product of coconut processing.

    Science.gov (United States)

    Rodsamran, Pattrathip; Sothornvit, Rungsinee

    2018-02-15

    Coconut cake, a by-product from milk and oil extractions, contains a high amount of protein. Protein extraction from coconut milk cake and coconut oil cake was investigated. The supernatant and precipitate protein powders from both coconut milk and oil cakes were compared based on their physicochemical and functional properties. Glutelin was the predominant protein fraction in both coconut cakes. Protein powders from milk cake presented higher water and oil absorption capacities than those from oil cake. Both protein powders from oil cake exhibited better foaming capacity and a better emulsifying activity index than those from milk cake. Coconut proteins were mostly solubilized in strong acidic and alkaline solutions. Minimum solubility was observed at pH 4, confirming the isoelectric point of coconut protein. Therefore, the coconut residues after extractions might be a potential alternative renewable plant protein source to use asa food ingredient to enhance food nutrition and quality. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Concentration and entry rate of amino acids in buffalo calves fed on two planes of crude protein

    International Nuclear Information System (INIS)

    Verma, D.N.; Singh, U.B.; Varma, A.; Ranjhan, S.K.

    1974-01-01

    Amino acid entry rates into the body pool have been estimated in buffalo calves using a single injection isotope dilution technique. The animals received 2 levels of crude protein, 13 percent lower and 19 percent higher than NRC recommendation. The concentrations of free amino acid in plasma were 5.49 and 7.17 mg/100 ml in animals fed on low and high crude protein diet, respectively. There was significant differences in the plasma amino acid concentration and entry rates between the groups. Amino acid entry rates were 79.17 and 117.78 mg per min in groups fed on low and high plane of crude protein respectively, showing that availability of amino acid is better in animals given ratio high in crude protein contents. (author)

  9. Day/night changes in serum S100B protein concentrations in acute paranoid schizophrenia.

    Science.gov (United States)

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

    2017-04-03

    There are day/night and seasonal changes in biological markers such as melatonin and cortisol. Controversial changes in serum S100B protein levels have been described in schizophrenia. We aim studying whether serum S100B levels present day/night variations in schizophrenia patients and whether S100B levels are related to psychopathology. Sixty-five paranoid schizophrenic inpatients participated in the study. Psychopathology was assessed with the Positive and Negative Syndrome Scale (PANSS) at admission and discharge. Blood was drawn at 12:00 (midday) and 00:00 (midnight) hours at admission and discharge. Sixty-five healthy subjects matched by age, gender and season acted as control group. At admission and discharge patients had significantly higher serum S100B concentrations at midday and midnight than healthy subjects. At admission, patients showed a day/night variation of S100B levels, with higher S100B levels at 12:00 than at 00:00h (143.7±26.3pg/ml vs. 96.9±16.6pg/ml). This day/night difference was not present in the control group. Midday and midnight S100B at admission decreased when compared to S100B at discharge (midday, 143.7±26.3 vs. 83.0±12, midnight 96.9±16.6 vs. 68.6±14.5). There was a positive correlation between the PANSS positive subscale and S100B concentrations at admission. This correlation was not present at discharge. acute paranoid schizophrenia inpatients present a day/night change of S100B serum levels at admission that disappears at discharge. The correlation between serum S100B concentrations and the PANSS positive scores at admission as well as the decrease of S100B at discharge may be interpreted as an acute biological response to the clinical state of the patients. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  10. Feed efficiency of diets with different energy and protein concentrations supplemented with methionine in laying quails

    Science.gov (United States)

    Ratriyanto, A.; Indreswari, R.; Nuhriawangsa, A. M. P.; Purwanti, E.

    2018-03-01

    The study was conducted to evaluate the feed efficiency of quail diets containing different concentrations of metabolizable energy (ME) and crude protein (CP) with constant ratio and supplemented with methionine. Four hundred laying quails (Coturnix coturnix japonica) were randomly assigned to four experimental diets in a 2×2 factorial arrangement. Each dietary treatment used 5 replicates of 20 quails. Two basal diets were formulated to contain 2,800 kcal kg-1 ME and 18.7% CP (High ME-CP) and 2,600 kcal kg-1 ME and 17.3% CP (Low ME-CP). Each basal diet was supplemented with 0 and 0.12% methionine. The High ME-CP diets generated lower feed consumption but higher egg mass and feed efficiency (Pprotein efficiency ratio (PHigh ME-CP supplemented with methionine resulted the highest feed efficiency followed by the Low ME-CP supplemented with methionine, while both High ME-CP and Low ME-CP without methionine supplementation resulted the lowest feed efficiency (PHigh ME-CP supplemented with 0.12% methionine provided benefit to improve the feed efficiency in laying quails.

  11. Whey protein phospholipid concentrate and delactosed permeate: Applications in caramel, ice cream, and cake.

    Science.gov (United States)

    Levin, M A; Burrington, K J; Hartel, R W

    2016-09-01

    Whey protein phospholipid concentrate (WPPC) and delactosed permeate (DLP) are 2 coproducts of cheese whey processing that are currently underutilized. Past research has shown that WPPC and DLP can be used together as a functional dairy ingredient in foods such as ice cream, soup, and caramel. However, the scope of the research has been limited to a single WPPC supplier. The variability of the composition and functionality of WPPC was previously studied. The objective of this research was to expand on the previous study and examine the potential applications of WPPC and DLP blends in foods. In ice cream, WPPC was added as a natural emulsifier to replace synthetic emulsifiers. The WPPC decreased the amount of partially coalesced fat and increased the drip-through rate. In caramel, DLP and WPPC replaced sweetened condensed skim milk and lecithin. Cold flow increased significantly, and hardness and stickiness decreased. In cake, DLP and WPPC were added as a total replacement of eggs, with no change in yield, color, or texture. Overall, WPPC and DLP can be utilized as functional dairy ingredients at a lower cost in ice cream and cake but not in chewy caramel. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  12. Effects of carprofen and meloxicam on C-reactive protein, ceruloplasmin, and fibrinogen concentrations in dogs undergoing ovariohysterectomy.

    Science.gov (United States)

    Kum, Cavit; Voyvoda, Huseyin; Sekkin, Selim; Karademir, Umit; Tarimcilar, Tugrul

    2013-10-01

    To evaluate the effects of perioperative oral administration of carprofen and meloxicam on concentrations of 3 acute-phase proteins in dogs undergoing elective ovariohysterectomy (OVH). 18 healthy adult anestrous female dogs undergoing elective OVH. Dogs were allocated to 3 groups (6 dogs/group). A placebo treatment, carprofen (2.0 mg/kg), or meloxicam (0.2 mg/kg) was orally administered to the dogs of the respective groups. The initial doses were administered 30 minutes before premedication prior to OVH; additional doses were administered once daily for 4 days after surgery. Blood samples were collected 45 minutes before premedication and 4, 8, 12, 24, 36, 48, 72, 96, and 120 hours after the end of OVH; samples were used for measurement of total WBC and neutrophil counts and concentrations of C-reactive protein (CRP), ceruloplasmin, and fibrinogen. Values did not differ significantly among groups for WBC and neutrophil counts, serum concentrations of CRP and ceruloplasmin, and plasma concentrations of fibrinogen. Concentrations of all inflammatory markers, except serum ceruloplasmin, increased significantly following OVH, but in a similar manner for each group. No significant changes were detected in serum ceruloplasmin concentrations over time. Perioperative administration of both carprofen and meloxicam did not significantly affect the concentrations of CRP, ceruloplasmin, and fibrinogen in dogs undergoing OVH. Thus, use of carprofen or meloxicam should not affect clinical interpretation of results for these 3 acute-phase proteins.

  13. Signaling Pathways Related to Protein Synthesis and Amino Acid Concentration in Pig Skeletal Muscles Depend on the Dietary Protein Level, Genotype and Developmental Stages.

    Science.gov (United States)

    Liu, Yingying; Li, Fengna; Kong, Xiangfeng; Tan, Bie; Li, Yinghui; Duan, Yehui; Blachier, François; Hu, Chien-An A; Yin, Yulong

    2015-01-01

    Muscle growth is regulated by the homeostatic balance of the biosynthesis and degradation of muscle proteins. To elucidate the molecular interactions among diet, pig genotype, and physiological stage, we examined the effect of dietary protein concentration, pig genotype, and physiological stages on amino acid (AA) pools, protein deposition, and related signaling pathways in different types of skeletal muscles. The study used 48 Landrace pigs and 48 pure-bred Bama mini-pigs assigned to each of 2 dietary treatments: lower/GB (Chinese conventional diet)- or higher/NRC (National Research Council)-protein diet. Diets were fed from 5 weeks of age to respective market weights of each genotype. Samples of biceps femoris muscle (BFM, type I) and longissimus dorsi muscle (LDM, type II) were collected at nursery, growing, and finishing phases according to the physiological stage of each genotype, to determine the AA concentrations, mRNA levels for growth-related genes in muscles, and protein abundances of mechanistic target of rapamycin (mTOR) signaling pathway. Our data showed that the concentrations of most AAs in LDM and BFM of pigs increased (Prelated AA, including Met, Phe, Tyr, Pro, and Ser, compared with Landrace pigs. The mRNA levels for myogenic determining factor, myogenin, myocyte-specific enhancer binding factor 2 A, and myostatin of Bama mini-pigs were higher (P<0.05) than those of Landrace pigs, while total and phosphorylated protein levels for protein kinase B, mTOR, and p70 ribosomal protein S6 kinases (p70S6K), and ratios of p-mTOR/mTOR, p-AKT/AKT, and p-p70S6K/p70S6K were lower (P<0.05). There was a significant pig genotype-dependent effect of dietary protein on the levels for mTOR and p70S6K. When compared with the higher protein-NRC diet, the lower protein-GB diet increased (P<0.05) the levels for mTOR and p70S6K in Bama mini-pigs, but repressed (P<0.05) the level for p70S6K in Landrace pigs. The higher protein-NRC diet increased ratio of p-mTOR/mTOR in

  14. Protein and starch concentrates of air-classified field pea and zero-tannin faba bean for weaned pigs.

    Science.gov (United States)

    Gunawardena, C K; Zijlstra, R T; Goonewardene, L A; Beltranena, E

    2010-08-01

    Air-classified pulse (non-oilseed legume) protein and starch may replace specialty protein and starch feedstuffs in diets for weaned pigs. In Exp. 1, three specialty protein sources (5% soy protein concentrate, 5% corn gluten meal, and 5% menhaden meal in the control diet) were replaced with 16% zero-tannin hulled or dehulled faba bean, or 17.5% field pea protein concentrate. In total, 192 group-housed pigs (2 gilts and 2 barrows per pen; BW = 7.5 +/- 1.4 kg) were fed wheat-based diets (3.60 Mcal/kg of DE and 3.3 g of standardized ileal digestible Lys/Mcal DE) over 28 d for 12 pen observations per each of 4 diets. Overall, protein source did not affect ADFI, ADG, or G:F. Apparent total tract digestibility (ATTD) of DM, GE, and P was greater (P bean and field pea protein concentrate diets than the diet with 3 specialty protein sources. In Exp. 2, faba bean and field pea starch concentrates were compared with corn, wheat, tapioca, and potato starch as dietary energy sources. In total, 36 individually housed barrows (BW = 8.0 +/- 1.5 kg) were fed 1 of 6 diets for 15 d. Feces and urine were collected from d 8 to 14, and jugular blood was sampled after overnight fast and refeeding on d 15. Starch source did not affect N retention as a percentage of N intake. For d 0 to 14, ADFI of pigs fed field pea starch was greater (P bean starch. Pigs fed tapioca, field pea, wheat, or corn starch grew faster (P bean or potato starch. For d 0 to 14, pigs fed corn or wheat starch had a 0.1 greater (P bean, field pea, or potato starch. The ATTD of DM, GE, CP, and starch and the DE value of potato starch were much less (P bean, wheat, and potato starch, respectively. However, postprandial plasma insulin tended to be 844 and 577 pmol/L greater (P bean and corn starch, respectively, than for pigs fed potato starch. The high insulin response of pigs fed faba starch could not be explained. In conclusion, air-classified pulse protein concentrates can replace specialty protein feedstuffs in

  15. Effects of dietary starch and protein levels on milk production and composition of dairy cows fed high concentrate diet

    Directory of Open Access Journals (Sweden)

    Mustafa Güçlü Sucak

    2017-07-01

    Full Text Available Abstract Twenty eight Holstein cows (averaged 41±31.5 and 82±24 days in milk, and 30.4±3.49 and 29.0±2.22 kg/d milk yield were fed a high concentrate diet (70:30 concentrate to forage to examine effects on milk production and composition. The cows were randomly assigned to receive four dietary treatments according to a 2 x 2 factorial arrangement. Factors were starch (14% and 22% and protein (15% and 18%. Wheat straw was used as forage source. The study lasted 6 weeks. Dry matter intake was not affected (P> 0.05 by the dietary treatments in the study. Milk yield increased with increased dietary protein level (P< 0.01. Milk urea nitrogen concentrations were affected by dietary protein and starch levels, but there was no interaction effect. Nitrogen efficiency (Milk N/N intake was decreased by increasing in dietary protein level (P< 0.01. In conclusion, the cows fed total mixed ration (TMR containing low level of wheat straw responded better when dietary protein increased. But, efficiency of N use and N excretion to the environment were worsened. Key words: Dairy cattle, milk composition, protein, starch, wheat straw

  16. Hippocampal kindling alters the concentration of glial fibrillary acidic protein and other marker proteins in rat brain

    DEFF Research Database (Denmark)

    Hansen, A; Jørgensen, Ole Steen; Bolwig, T G

    1990-01-01

    The effect of hippocampal kindling on neuronal and glial marker proteins was studied in the rat by immunochemical methods. In hippocampus, pyriform cortex and amygdala there was an increase in glial fibrillary acidic protein (GFAP), indicating reactive gliosis, and an increase in the glycolytic...... enzyme NSE, suggesting increased anaerobic metabolism. Neuronal cell adhesion molecule (NCAM) decreased in pyriform cortex and amygdala of kindled rats, indicating neuronal degeneration....

  17. Predicting the activity coefficients of free-solvent for concentrated globular protein solutions using independently determined physical parameters.

    Directory of Open Access Journals (Sweden)

    Devin W McBride

    Full Text Available The activity coefficient is largely considered an empirical parameter that was traditionally introduced to correct the non-ideality observed in thermodynamic systems such as osmotic pressure. Here, the activity coefficient of free-solvent is related to physically realistic parameters and a mathematical expression is developed to directly predict the activity coefficients of free-solvent, for aqueous protein solutions up to near-saturation concentrations. The model is based on the free-solvent model, which has previously been shown to provide excellent prediction of the osmotic pressure of concentrated and crowded globular proteins in aqueous solutions up to near-saturation concentrations. Thus, this model uses only the independently determined, physically realizable quantities: mole fraction, solvent accessible surface area, and ion binding, in its prediction. Predictions are presented for the activity coefficients of free-solvent for near-saturated protein solutions containing either bovine serum albumin or hemoglobin. As a verification step, the predictability of the model for the activity coefficient of sucrose solutions was evaluated. The predicted activity coefficients of free-solvent are compared to the calculated activity coefficients of free-solvent based on osmotic pressure data. It is observed that the predicted activity coefficients are increasingly dependent on the solute-solvent parameters as the protein concentration increases to near-saturation concentrations.

  18. Stable isotopes

    International Nuclear Information System (INIS)

    Evans, D.K.

    1986-01-01

    Seventy-five percent of the world's stable isotope supply comes from one producer, Oak Ridge Nuclear Laboratory (ORNL) in the US. Canadian concern is that foreign needs will be met only after domestic needs, thus creating a shortage of stable isotopes in Canada. This article describes the present situation in Canada (availability and cost) of stable isotopes, the isotope enrichment techniques, and related research programs at Chalk River Nuclear Laboratories (CRNL)

  19. Variances in nutrient content and yield of alfalfa protein concentrate processed with five methods

    Science.gov (United States)

    The demand for protein is growing with increased populations and world affluence. A sustainable and affordable protein source is needed to support the growing aquaculture industry worldwide. Alfalfa produces high levels of protein and provides numerous environmental services, potentially making it a...

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

    DEFF Research Database (Denmark)

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

    2013-01-01

    Objective-To evaluate associations of serum C-reactive protein (CRP) concentration with duration of hospitalization and with outcome in puppies with canine parvoviral enteritis. Design-Prospective observational study. Animals-79 client-owned puppies with naturally acquired canine parvovirus...... infection. Procedures-All puppies received supportive care. Serum CRP concentration was measured at the time of admission, approximately every 10 to 12 hours for the first 48 hours, and then every 24 hours until discharge from the hospital or death. Associations between outcome and CRP concentration...... at various time points or changes in CRP concentration over time were assessed via multiple logistic regression. Associations of CRP concentration with survival time and duration of hospitalization among survivors were estimated with Cox proportional hazards regression. Use of CRP concentration to predict...

  1. Application of gamma radiation and heat treatment for the preparation of fish protein concentrates from Bombay duck (Harpodon nehereus)

    International Nuclear Information System (INIS)

    Warrier, S.B.K.; Ninjoor, V.; Kumta, U.S.

    1976-01-01

    Limitations in the conventional procedures for the manufacture of fish protein concentrate (FPG), such as residual toxicity of the solvents and loss in functional properties necessitate the need for evolving newer methods. A simple method has been developed for the preparation of FPC from the muscle of Bombay duck employing radiation-heat combination treatments. This involves extraction of the muscle proteins in 0.01 M phosphate buffer containing 5 percent sodium chloride, pH 7.5, irradiation at a dose of 100-250 krad followed by heat treatment at 60 degC for 10 min. The procedure permits the precipitation of fibrillar proteins to the extent of 83 percent. There is no change in the digestibility of the precipitated proteins as revealed by the tryptic hydrolysis. These results point to the synergestic effect of radiation and heat on the precipitation of proteins - an observation that has high potential for scaling up as a new process. (author)

  2. Glucose, fructose and sucrose increase the solubility of protein-tannin complexes and at high concentration, glucose and sucrose interfere with bisulphite bleaching of wine pigments.

    Science.gov (United States)

    Harbertson, James F; Yuan, Chunlong; Mireles, Maria S; Hanlin, Rachel L; Downey, Mark O

    2013-05-01

    Wines were modified with increasing sugar concentrations and decreasing tannin concentrations and analysed by a combination of protein precipitation and bisulphite bleaching. Increasing sugar concentration decreased the precipitation of tannin and protein-precipitable polymeric pigments (PPP). The use of a hydrogen bond disruptor (urea) to reduce protein-tannin and protein-pigment complex formation showed that the effect of sugar concentration occurred by increasing the solubility of the tannin-protein complex, not by interfering with protein-tannin complex formation. By increasing the solubility of pigment-protein complexes, non-protein-precipitable polymeric pigments (nPPP) appeared to increase. There was also an increase in total polymeric pigments at each tannin concentration with increasing glucose and sucrose concentration, indicating that sugar concentration might also affect bisulphite bleaching of wine pigments. While a significant effect of sugar concentration on tannin-protein complex solubility was observed, these effects were greatest at sugar concentrations far in excess of normal wine making conditions. Under normal wine making conditions, sugar concentration will have a negligible effect on protein-precipitable tannin, PPP and nPPP concentrations. Copyright © 2012 Elsevier Ltd. All rights reserved.

  3. Estimation of daily protein intake based on spot urine urea nitrogen concentration in chronic kidney disease patients.

    Science.gov (United States)

    Kanno, Hiroko; Kanda, Eiichiro; Sato, Asako; Sakamoto, Kaori; Kanno, Yoshihiko

    2016-04-01

    Determination of daily protein intake in the management of chronic kidney disease (CKD) requires precision. Inaccuracies in recording dietary intake occur, and estimation from total urea excretion presents hurdles owing to the difficulty of collecting whole urine for 24 h. Spot urine has been used for measuring daily sodium intake and urinary protein excretion. In this cross-sectional study, we investigated whether urea nitrogen (UN) concentration in spot urine can be used to predict daily protein intake instead of the 24-h urine collection in 193 Japanese CKD patients (Stages G1-G5). After patient randomization into 2 datasets for the development and validation of models, bootstrapping was used to develop protein intake estimation models. The parameters for the candidate multivariate regression models were male gender, age, body mass index (BMI), diabetes mellitus, dyslipidemia, proteinuria, estimated glomerular filtration rate, serum albumin level, spot urinary UN and creatinine level, and spot urinary UN/creatinine levels. The final model contained BMI and spot urinary UN level. The final model was selected because of the higher correlation between the predicted and measured protein intakes r = 0.558 (95 % confidence interval 0.400, 0.683), and the smaller distribution of the difference between the measured and predicted protein intakes than those of the other models. The results suggest that UN concentration in spot urine may be used to estimate daily protein intake and that a prediction formula would be useful for nutritional control in CKD patients.

  4. The use of protein structure/activity relationships in the rational design of stable particulate delivery systems

    Directory of Open Access Journals (Sweden)

    M.H.B. Costa

    2002-06-01

    Full Text Available The recombinant heat shock protein (18 kDa-hsp from Mycobacterium leprae was studied as a T-epitope model for vaccine development. We present a structural analysis of the stability of recombinant 18 kDa-hsp during different processing steps. Circular dichroism and ELISA were used to monitor protein structure after thermal stress, lyophilization and chemical modification. We observed that the 18 kDa-hsp is extremely resistant to a wide range of temperatures (60% of activity is retained at 80ºC for 20 min. N-Acylation increased its ordered structure by 4% and decreased its ß-T1 structure by 2%. ELISA demonstrated that the native conformation of the 18 kDa-hsp was preserved after hydrophobic modification by acylation. The recombinant 18 kDa-hsp resists to a wide range of temperatures and chemical modifications without loss of its main characteristic, which is to be a source of T epitopes. This resistance is probably directly related to its lack of organization at the level of tertiary and secondary structures.

  5. Concentration of total proteins in blood plasma of chickens hatched from irradiated eggs with low dose gamma radiation

    International Nuclear Information System (INIS)

    Vilic, M.; Kraljevic, P.; Miljanic, S.; Simpraga, M.

    2005-01-01

    It is known that low-dose ionising radiation may have stimulating effects on chickens. Low doses may also cause changes in the concentration of blood plasma total proteins, glucose and cholesterol in chickens. This study investigates the effects of low dose gamma-radiation on the concentration of total proteins in the blood plasma of chickens hatched from eggs irradiated with a dose of 0.15 Gy on incubation days 7 and 19. Results were compared with the control group (chickens hatched from non-irradiated eggs). After hatching, all other conditions were the same for both groups. Blood samples were drawn from the heart, and later from the wing vein on days 1, 3, 5, 7,10, 20, 30 and 42. The concentration of total proteins was determined spectrophotometrically using Boehringer Mannheim GmbH optimised kits. The concentration of total proteins in blood plasma in chickens hatched from eggs irradiated with 0.15 Gy on incubation day 7 showed a statistically significant decrease on the sampling day 3 (P less than 0.05) and 7 (P less than 0.01). The concentration of total proteins in blood plasma in chickens hatched from eggs irradiated with 0.15 Gy on incubation day 19 showed a statistically significant increase only on sampling day 1 (P less than 0.05). These results suggest that exposure of eggs to 0.15 Gy of gamma-radiation on the 7th and 19th day of incubation could produce different effects on the protein metabolism in chickens.(author)

  6. Chemical composition of protein concentrate prepared from Yellowfin tuna Thunnus albacares roe by cook-dried process

    Directory of Open Access Journals (Sweden)

    Hyun Ji Lee

    2016-05-01

    Full Text Available Abstract Roe is the term used to describe fish eggs (oocytes gathered in skeins and is one of the most valuable food products from fishery sources. Thus, means of processing are required to convert the underutilized yellowfin tuna roes (YTR into more marketable and acceptable forms as protein concentrate. Roe protein concentrates (RPCs were prepared by cooking condition (boil-dried concentrate, BDC and steam-dried concentrate, SDC, respectively and un-cooking condition (freeze-dried concentrate, FDC from yellowfin tuna roe. The yield of RPCs was in the range from 22.2 to 25.3 g/100 g of roe. RPCs contained protein (72.3–77.3 %, moisture (4.3–5.6 %, lipid (10.6–11.3 % and ash (4.3–5.7 % as the major constituents. The prominent amino acids of RPCs were aspartic acid, 8.7–9.2, glutamic acid, 13.1–13.2, and leucine, 8.5–8.6 g/100 g of protein. Major differences were not observed in each of the amino acid. K, S, Na, and P as minerals were the major elements in RPCs. No difference noted in sodium dodecyl sulfate polyacrylamide gel electrophoresis protein band (15–100 K possibly representing partial hydrolysis of myosin. Therefore, RPCs from YTR could be use potential protein ingredient for human food and animal feeds.

  7. Vitamin D-binding protein and free vitamin D concentrations in acromegaly.

    Science.gov (United States)

    Altinova, Alev Eroglu; Ozkan, Cigdem; Akturk, Mujde; Gulbahar, Ozlem; Yalcin, Muhittin; Cakir, Nuri; Toruner, Fusun Balos

    2016-05-01

    Free 25-hydroxyvitamin D [25(OH)D] is suggested to be important in the determination of vitamin D deficiency, since vitamin D-binding protein (VDBP) may affect total 25(OH)D levels. There are no data about free 25(OH)D concentrations in acromegaly. We aimed to investigate serum VDBP and total and free 25(OH)D levels in patients with acromegaly in comparison with control subjects. We recruited 54 patients with acromegaly and 32 control subjects who were similar according to age, gender, and body mass index. Serum VDBP levels were found to be increased in patients with acromegaly compared to control subjects [90.35 (72.45-111.10) vs. 69.52 (63.89-80.13) mg/l, p = 0.001]. There was statistically no significant difference in serum total 25(OH)D levels between the patients with acromegaly and control subjects [18.63 (13.35-27.73) vs. 22.51 (19.20-28.96) ng/ml, p = 0.05]. Free 25(OH)D levels were significantly decreased in patients with acromegaly compared to control subjects [14.55 (10.45-21.45) vs. 17.75 (15.30-23.75) pg/ml, p = 0.03]. Free 25(OH)D levels correlated positively with total 25(OH)D (p = 0.0001) and HDL cholesterol (p = 0.04) and negatively with fasting blood glucose (p = 0.04). Our findings indicate that VDBP is increased and free 25(OH)D is decreased in acromegaly, while there is no significant alteration in total 25(OH)D.

  8. Extensive characterization of a lentiviral-derived stable cell line expressing rabbit hemorrhagic disease virus VPg protein.

    Science.gov (United States)

    Zhu, Jie; Miao, Qiuhong; Tan, Yonggui; Guo, Huimin; Li, Chuanfeng; Chen, Zongyan; Liu, Guangqing

    2016-11-01

    Rabbit hemorrhagic disease virus (RHDV) is an important member of the caliciviridae family. Currently, no suitable tissue culture system is available for proliferating RHDV, which limits the study of its pathogenesis. To bypass this obstacle, we established a cell line, RK13-VPg, stably expressing the VPg gene with a lentivirus packaging system in this study. In addition, the recently constructed RHDV replicon in our laboratory provided an appropriate model for studying the pathogenesis of RHDV without in vitro RHDV propagation and culture. Using this RHDV replicon and RK13-VPg cell line, we further demonstrated that the presence of VPg protein is essential for efficient translation of an RHDV replicon. Therefore, the RK13-VPg cell line is a powerful tool for studying the replication and translation mechanisms of RHDV. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Involvement of protein kinase C in the mechanism of action of Escherichia coli heat-stable enterotoxin (STa) in a human colonic carcinoma cell line, COLO-205

    International Nuclear Information System (INIS)

    Gupta, Dyuti Datta; Saha, Subhrajit; Chakrabarti, Manoj K.

    2005-01-01

    The present study was undertaken to determine the involvement of calcium-protein kinase C pathway in the mechanism of action of Escherichia coli heat stable enterotoxin (STa) apart from STa-induced activation of guanylate cyclase in human colonic carcinoma cell line COLO-205, which was used as a model cultured cell line to study the mechanism of action of E. coli STa. In response to E. coli STa, protein kinase C (PKC) activity was increased in a time-dependent manner with its physical translocation from cytosol to membrane. Inhibition of the PKC activity in membrane fraction and inhibition of its physical translocation in response to IP 3 -mediated calcium release inhibitor dantrolene suggested the involvement of intracellular store depletion in the regulation of PKC activity. Among different PKC isoforms, predominant involvement of calcium-dependent protein kinase C (PKCα) was specified using isotype-specific pseudosubstrate, which showed pronounce enzyme activity. Inhibition of enzyme activity by PKCα-specific inhibitor Goe6976 and immunoblott study employing isotype-specific antibody further demonstrated the involvement of calcium-dependent isoform of PKC in the mechanism of action of E. coli STa. Moreover, inhibition of guanylate cyclase activity by PKCα-specific inhibitor Goe6976 suggested the involvement of PKCα in the regulation of guanylate cyclase activity

  10. Lah is a transmembrane protein and requires Spa10 for stable positioning of Woronin bodies at the septal pore of Aspergillus fumigatus.

    Science.gov (United States)

    Leonhardt, Yannik; Kakoschke, Sara Carina; Wagener, Johannes; Ebel, Frank

    2017-03-10

    Woronin bodies are specialized, fungal-specific organelles that enable an immediate closure of septal pores after injury to protect hyphae from excessive cytoplasmic bleeding. In most Ascomycetes, Woronin bodies are tethered at the septal pore by so-called Lah proteins. Using the pathogenic mold Aspergillus fumigatus as a model organism, we show that the C-terminal 288 amino acids of Lah (LahC 288 ) bind to the rim of the septal pore. LahC 288 essentially consists of a membrane spanning region and a putative extracellular domain, which are both required for the targeting to the septum. In an A. fumigatus rho4 deletion mutant that has a severe defect in septum formation, LahC 288 is recruited to spot-like structures in or at the lateral membrane. This suggests that LahC is recruited before Rho4 starts to govern the septation process. Accordingly, we found that in wild type hyphae Lah is bound before a cross-wall emerges and thus enables a tethering of Woronin bodies at the site of the newly formed septum. Finally, we identified Spa10, a member of a recently described family of septal pore-associated proteins, as a first protein that directly or indirectly interacts with LahC to allow a stable positioning of Woronin bodies at the mature septum.

  11. Investigation of mercury-containing proteins by enriched stable isotopic tracer and size-exclusion chromatography hyphenated to inductively coupled plasma-isotope dilution mass spectrometry

    International Nuclear Information System (INIS)

    Shi Junwen; Feng Weiyue; Wang Meng; Zhang Fang; Li Bai; Wang Bing; Zhu Motao; Chai Zhifang

    2007-01-01

    In order to investigate trace mercury-containing proteins in maternal rat and their offspring, a method of enriched stable isotopic tracer ( 196 Hg and 198 Hg) combined with size-exclusion chromatography (SEC) coupled to inductively coupled plasma-isotope dilution mass spectrometry (ICP-IDMS) was developed. Prior to the analysis, 196 Hg- and 198 Hg-enriched methylmercury was administrated to the pregnant rats. Then the mercury-containing proteins in serum and brain cytosol of the dam and pup rats were separated by size-exclusion columns and the mercury was detected by ICP-MS. The ICP-MS spectrogram of the tracing samples showed significantly elevated 196 Hg and 198 Hg isotopic signals compared with the natural ones, indicating that the detection sensitivity could be increased by the tracer method. The contents of mercury in chromatographic fractions of the dam and pup rat brain cytosol were quantitatively estimated by post-column reverse ID-ICP-MS. The quantitative speciation differences of mercury in brain cytosol between the dam and pup rats were observed, indicating that such studies could be useful for toxicological estimation. Additionally, the isotopic ratio measurement of 198 Hg/ 202 Hg in the tracing samples could be used to identify the artifact mercury species caused in the analytical procedure. The study demonstrates that the tracer method combined with high-performance liquid chromatography (HPLC)-ICP-IDMS could provide reliably qualitative and quantitative information on mercury-containing proteins in organisms

  12. Grain protein concentration and harvestable protein under future climate conditions. A study of 108 spring barley accessions

    DEFF Research Database (Denmark)

    Ingvordsen, Cathrine Heinz; Gislum, René; Jørgensen, Johannes Ravn

    2016-01-01

    In the present study a set of 108 spring barley (H. vulgare L.) accessions were cultivated under predicted future levels of temperature and [CO2] as single factors and in combination (IPCC, AR5, RCP8.5). Across all genotypes, elevated [CO2] (700 ppm day/night) slightly decreased protein concentra......In the present study a set of 108 spring barley (H. vulgare L.) accessions were cultivated under predicted future levels of temperature and [CO2] as single factors and in combination (IPCC, AR5, RCP8.5). Across all genotypes, elevated [CO2] (700 ppm day/night) slightly decreased protein...

  13. The protein concentration of blood coagulation factor VII can be measured equally well in plasma and serum

    DEFF Research Database (Denmark)

    Bladbjerg, E-M; Overgaard, K; Gram, J

    1995-01-01

    In the Northwick Park Heart Study, the coagulant activity of factor VII (FVII:C) has been identified as a risk marker of ischaemic heart disease. In the fasting state, the protein concentration of FVII (FVII:Ag) might be an even better risk marker, because of the low coefficient of variation...

  14. The effects of high levels of rumen degradable protein on rumen pH and histamine concentrations in dairy cows

    NARCIS (Netherlands)

    Pilachai, R.; Schonewille, J.T.; Thamrongyoswittayakul, C.; Aiumlamai, S.; Wachirapakom, C.; Everts, H.; Hendriks, W.H.

    2012-01-01

    An experiment was conducted to test the hypothesis that the supplementation of crude protein (CP) results in rumen acidosis and increased histamine concentrations in dairy cows. Six ruminally fistulated, non-pregnant dry cows were fed three experimental rations in a replicated 3 × 3 Latin square

  15. Determination of the Influence of Substrate Concentration on Enzyme Selectivity Using Whey Protein Isolate and Bacillus licheniformis Protease

    NARCIS (Netherlands)

    Butré, C.I.; Sforza, S.; Gruppen, H.; Wierenga, P.A.

    2014-01-01

    Increasing substrate concentration during enzymatic protein hydrolysis results in a decrease in hydrolysis rate. To test if changes in the mechanism of hydrolysis also occur, the enzyme selectivity was determined. The selectivity is defined quantitatively as the relative rate of hydrolysis of each

  16. The effect of heparin on pregnancy associated plasma protein-A concentration in healthy, non-pregnant individuals

    DEFF Research Database (Denmark)

    Jespersen, Camilla H B; Vestergaard, Kirstine R.; Schou, Morten

    2015-01-01

    Objectives: The objective of this study was to determine the differences in pregnancy associated plasma protein-A (PAPP-A) concentrations in heparin naive and heparin treated healthy men and non-pregnant women, to find a possible difference in different age groups, and to determine the response...

  17. Conformational Analysis of Proteins in Highly Concentrated Solutions by Dialysis-Coupled Hydrogen/Deuterium Exchange Mass Spectrometry

    DEFF Research Database (Denmark)

    Houde, Damian; Esmail Nazari, Zeinab; Bou-Assaf, George M

    2016-01-01

    When highly concentrated, an antibody solution can exhibit unusual behaviors, which can lead to unwanted properties, such as increased levels of protein aggregation and unusually high viscosity. Molecular modeling, along with many indirect biophysical measurements, has suggested that the cause fo...... in industry. Graphical Abstract ᅟ....

  18. Introducing enzyme selectivity as a quantitative parameter to describe the effects of substrate concentration on protein hydrolysis

    NARCIS (Netherlands)

    Butré, C.I.

    2014-01-01

    To understand the differences in peptide composition that result from variations in the conditions of enzymatic hydrolysis of proteins (e.g. substrate concentration) the mechanism of hydrolysis needs to be understood in detail. Therefore, methods and tools were developed to characterize and

  19. The effect of concentrate in meal- or pellet form on milk, protein and ...

    African Journals Online (AJOL)

    Keywords: Low butterfat syndrome, dairy concentrate pellets. The feeding of concentrate pellets has become general practice on many dairy farms in South ... (1 week for adaptation and 4 weeks for production). All ingredients (for the meal and ...

  20. Identification of Differentially Expressed Proteins in Liver in Response to Subacute Ruminal Acidosis (SARA Induced by High-concentrate Diet

    Directory of Open Access Journals (Sweden)

    X. Y. Jiang

    2014-08-01

    Full Text Available The aim of this study was to evaluate protein expression patterns of liver in response to subacute ruminal acidosis (SARA induced by high-concentrate diet. Sixteen healthy mid-lactating goats were randomly divided into 2 groups and fed either a high-forage (HF diet or a high-concentrate (HC diet. The HC diet was expected to induce SARA. After ensuring the occurrence of SARA, liver samples were collected. Proteome analysis with differential in gel electrophoresis technology revealed that, 15 proteins were significantly modulated in liver in a comparison between HF and HC-fed goats. These proteins were found mainly associated with metabolism and energy transfer after identified by matrix-assisted laser desorption ionization/time of flight. The results indicated that glucose, lipid and protein catabolism could be enhanced when SARA occurred. It prompted that glucose, lipid and amine acid in the liver mainly participated in oxidation and energy supply when SARA occurred, which possibly consumed more precursors involved in milk protein and milk fat synthesis. These results suggest new candidate proteins that may contribute to a better understanding of the mechanisms that mediate liver adaptation to SARA.

  1. Toxic effects of zinc from trout farm sediments on ATP, protein, and hemoglobin concentrations of Limnodrilus hoffmeisteri.

    Science.gov (United States)

    Martinez-Tabche, L; Gutiérrez Cabrera, I; Gómez Oliván, L; Galar Martinez, M; Germán Faz, C

    2000-04-14

    Zinc (Zn) is a nutritionally essential metal, and deficiency results in severe health consequences to aquatic organisms. In this study toxicity data for Limnodrilus hoffmeisteri produced by Zn in systems using three natural sediments (trout farms: El Oyamel, El Truchón, and El Potrero) are presented. Hemoglobin, adenosine triphosphate (ATP), and protein concentrations were measured in L. hoffmeisteri exposed to spiked sediments, as indicators of exposure. Physicochemical characteristics of water and sediments were also considered. Zn concentrations were measured in water and sediment. El Oyamel, El Truchón, and El Potrero pond sediments did not have similar physicochemical characteristics. Zn concentrations of water obtained from the rustic ponds were near 0.4575 mg/L; however, this metal was always found to be higher in the sediments (0.0271-0.9754 mg/kg). The bioassay with worms demonstrated that pond sediments from El Oyamel, El Potrero, and El Truchón produced toxicity since ATP and protein concentrations were low compared to controls (organisms without metal). All spiked sediments had a significant reduction effect on ATP, protein, and hemoglobin concentrations. This investigation clearly shows that sediments of El Truchón, El Oyamel, and El Potrero possess toxicity potential. These results suggest the usefulness of these bioassays to evaluate the toxicity of sediments polluted with heavy metals.

  2. Recombinant protein of heptad-repeat HR212, a stable fusion inhibitor with potent anti-HIV action in vitro

    International Nuclear Information System (INIS)

    Pang, Wei; Wang Ruirui; Yang Liumeng; Liu Changmei; Tien Po; Zheng Yongtang

    2008-01-01

    HR212, a recombinant protein expressed in Escherichia coli, has been previously reported to inhibit HIV-1 membrane fusion at low nanomolar level. Here we report that HR212 is effective in blocking laboratory strain HIV-1 IIIB entry and replication with EC 50 values of 3.92 ± 0.62 and 6.59 ± 1.74 nM, respectively, and inhibiting infection by clinic isolate HIV-1 KM018 with EC 50 values of 44.44 ± 10.20 nM, as well as suppressing HIV-1-induced cytopathic effect with an EC 50 value of 3.04 ± 1.20 nM. It also inhibited HIV-2 ROD and HIV-2 CBL-20 entry and replication in the μM range. Notably, HR212 was highly effective against T20-resistant strains with EC 50 values ranging from 5.09 to 7.75 nM. Unlike T20, HR212 showed stability sufficient to inhibit syncytia formation in a time-of-addition assay, and was insensitive to proteinase K digestion. These results suggest that HR212 has great potential to be further developed as novel HIV-1 fusion inhibitor for treatment of HIV/AIDS patients, particularly for those infected by T20-resistant variants

  3. Cellular recovery from exposure to sub-optimal concentrations of AB toxins that inhibit protein synthesis

    Science.gov (United States)

    Shiga toxin 1, exotoxin A, diphtheria toxin and ricin are all AB-type protein toxins that act within the host cytosol to kill the host cell through a pathway involving the inhibition of protein synthesis. It is thought that a single molecule of cytosolic toxin is sufficient to kill the host cell. In...

  4. Effect of diet of Varying Protein concentrations on the Activity of ...

    African Journals Online (AJOL)

    Alterations in protein diet have been reported to result in alterations in calcium homeostasis in the body. Ca2+Mg2+ATPase is an ubiquitous enzyme important in calcium homeostasis in the body. The effect of varying protein diet on the activities of Ca2+ pump across cell membranes is however yet to be fully elucidated.

  5. Calorie restricted high protein diets downregulate lipogenesis and lower intrahepatic triglyceride concentrations in male rats

    Science.gov (United States)

    The purpose of this investigation was to assess the influence of calorie restriction (CR) alone, higher-protein/lower-carbohydrate intake alone, and combined CR higher-protein/lower-carbohydrate intake on glucose homeostasis, hepatic de novo lipogenesis (DNL), and intrahepatic triglycerides. Twelve-...

  6. Plasma intestinal fatty acid binding protein (I-FABP) concentrations increase following intestinal ischemia in pigs

    NARCIS (Netherlands)

    Niewold, T.A.; Meinen, M.; Meulen, van der J.

    2004-01-01

    Intestinal fatty acid binding protein (I-FABP) is an intracellular epithelial protein in the intestinal mucosa of many animals. IFABP appears in the circulation following epithelial damage, and in humans, is proven to be a parameter for damage to the mucosa. In this paper, an ELISA test designed for

  7. A robust method of determination of high concentrations of peptides and proteins

    DEFF Research Database (Denmark)

    Levashov, Pavel A; Sutherland, Duncan S; Besenbacher, Flemming

    2009-01-01

    In this paper, we pioneer application of a unique method of protein determination by coloring peptide bonds for analysis of a variety of biomolecules with different grades of purity (e.g., oligopeptides, membrane, and glycol proteins). We demonstrated that the calibration curve for all studied mo...

  8. Leaf Protein and Mineral Concentrations across the "Miracle Tree" Genus Moringa.

    Science.gov (United States)

    Olson, Mark E; Sankaran, Renuka P; Fahey, Jed W; Grusak, Michael A; Odee, David; Nouman, Wasif

    2016-01-01

    The moringa tree Moringa oleifera is a fast-growing, drought-resistant tree cultivated across the lowland dry tropics worldwide for its nutritious leaves. Despite its nutritious reputation, there has been no systematic survey of the variation in leaf nutritional quality across M. oleifera grown worldwide, or of the other species of the genus. To guide informed use of moringa, we surveyed protein, macro-, and micro- nutrients across 67 common garden samples of 12 Moringa taxa, including 23 samples of M. oleifera. Moringa oleifera, M. concanensis, M. stenopetala, an M. concanensis X oleifera hybrid, and M. longituba were highest in protein, with M. ruspoliana having the highest calcium levels. A protein-dry leaf mass tradeoff may preclude certain breeding possibilities, e.g. maximally high protein with large leaflets. These findings identify clear priorities and limitations for improved moringa varieties with traits such as high protein, calcium, or ease of preparation.

  9. Leaf Protein and Mineral Concentrations across the “Miracle Tree” Genus Moringa

    Science.gov (United States)

    Sankaran, Renuka P.; Fahey, Jed W.; Grusak, Michael A.; Odee, David; Nouman, Wasif

    2016-01-01

    The moringa tree Moringa oleifera is a fast-growing, drought-resistant tree cultivated across the lowland dry tropics worldwide for its nutritious leaves. Despite its nutritious reputation, there has been no systematic survey of the variation in leaf nutritional quality across M. oleifera grown worldwide, or of the other species of the genus. To guide informed use of moringa, we surveyed protein, macro-, and micro- nutrients across 67 common garden samples of 12 Moringa taxa, including 23 samples of M. oleifera. Moringa oleifera, M. concanensis, M. stenopetala, an M. concanensis X oleifera hybrid, and M. longituba were highest in protein, with M. ruspoliana having the highest calcium levels. A protein-dry leaf mass tradeoff may preclude certain breeding possibilities, e.g. maximally high protein with large leaflets. These findings identify clear priorities and limitations for improved moringa varieties with traits such as high protein, calcium, or ease of preparation. PMID:27459315

  10. Leaf Protein and Mineral Concentrations across the "Miracle Tree" Genus Moringa.

    Directory of Open Access Journals (Sweden)

    Mark E Olson

    Full Text Available The moringa tree Moringa oleifera is a fast-growing, drought-resistant tree cultivated across the lowland dry tropics worldwide for its nutritious leaves. Despite its nutritious reputation, there has been no systematic survey of the variation in leaf nutritional quality across M. oleifera grown worldwide, or of the other species of the genus. To guide informed use of moringa, we surveyed protein, macro-, and micro- nutrients across 67 common garden samples of 12 Moringa taxa, including 23 samples of M. oleifera. Moringa oleifera, M. concanensis, M. stenopetala, an M. concanensis X oleifera hybrid, and M. longituba were highest in protein, with M. ruspoliana having the highest calcium levels. A protein-dry leaf mass tradeoff may preclude certain breeding possibilities, e.g. maximally high protein with large leaflets. These findings identify clear priorities and limitations for improved moringa varieties with traits such as high protein, calcium, or ease of preparation.

  11. Influence of energy concentration and source on the utilization of feed protein and NPN in lambs. 2

    International Nuclear Information System (INIS)

    Ulbrich, M.; Bassuny, S.M.; Geissler, C.; Hoffmann, M.

    1989-01-01

    In an experiment 12 lambs of the species merino meet sheep were divided into 4 groups. The variants HE received 740 or 718 EFU cattle /kg DM and the variants NE 689 or 671 EFU cattle /kg DM. The different energy concentrations resulted from differentiated quotas of dried sugar beet chips and wheat starch supplements. Within the variants, sub-variants with (HESZ, NESZ) or without (HES, NES) sugar supplement were formed. Due to varied DM intake, the average energy intake in all groups was 42 EFU cattle /kg LW 0.75 . N balance experiments using 15 N-labelled urea were carried out, and 15 N accumulation of N excretion was projected to a steady state. The partial utilization of pure protein and NPN in the ration was ascertained with the help of a 3-pool compartment model of N utilization in ruminants. In the non-amino acid N pool HE utilized 84% of NPN and NE 77% for the synthesis of amino and nucleic acids. The efficiency of protein synthesis in the amino acid N-pool were in HESZ 64%, HES 70%, NESZ 70% and NES 73% resp. The total utilization of NPN is the sum of the partial utilization in the two pools, whereas the total utilization of pure protein is calculated from the true digestibility and the efficiency of the utilization in the AA-N pool. The total utilization of NPN for the synthesis of protein for the protein pool amounted to 40/35/41/33% and that of pure protein to 54/51/49/50%. Energy intake being identical, energy concentration did not have an influence on the utilization of pure protein and NPN, whereas NPN utilization was better in rations containing sugar. The pure protein in the ration was by 19 - 52% better utilized than NPN. (author)

  12. Serum C-reactive protein concentrations in Nova Scotia Duck Tolling Retrievers with immune-mediated rheumatic disease.

    Science.gov (United States)

    Bremer, Hanna Dorotea; Hillström, Anna; Kånåhols, Malin; Hagman, Ragnvi; Hansson-Hamlin, Helene

    2017-04-17

    Nova Scotia Duck Tolling Retrievers (NSDTRs) are a dog breed often affected by immune-mediated rheumatic disease (IMRD), a disorder characterised by chronic stiffness and joint pain. Most, but not all, dogs with IMRD, have antinuclear antibodies (ANA), which are also commonly present in the autoimmune disease systemic lupus erythematosus (SLE). The clinical and diagnostic findings of IMRD indicate that it is an SLE-related disorder. C-reactive protein (CRP), an acute phase protein, is a quantitative marker of inflammation for many diseases and is used for diagnosing and monitoring systemic inflammation in both humans and dogs. However, in human SLE, CRP concentrations are often elevated but correlate poorly with disease activity; they can be low in individual patients with active disease. The aim of the study was to investigate CRP in a group of NSDTRs with the SLE-related disorder IMRD. The hypothesis was that CRP concentrations would be increased in dogs with IMRD compared to healthy dogs, but that the increase would be mild. Serum CRP concentrations were measured in 18 IMRD-affected NSDTRs and 19 healthy control NSDTRs using two different canine-specific CRP assays. Dogs with IMRD and ANA had higher CRP concentrations than the control dogs, but the concentrations were below the clinical decision limit for systemic inflammation for most of the IMRD dogs. These results indicate that CRP concentrations were increased in dogs with IMRD and ANA, but the increase was mild, similar to what has been observed in human SLE.

  13. Technical Report on the Behavior of Trace Elements, Stable Isotopes, and Radiogenic Isotopes During the Processing of Uranium Ore to Uranium Ore Concentrate

    Energy Technology Data Exchange (ETDEWEB)

    Marks, N. E. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Borg, L. E. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Eppich, G. R. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Gaffney, A. M. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Genneti, V. G. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Hutcheon, I. D. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Kristo, M. J. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Lindvall, R. E. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Ramon, C. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Robel, M. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Roberts, S. K. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Schorzman, K. C. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Sharp, M. A. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Singleton, M. J. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Williams, R. W. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)

    2015-07-09

    The goals of this SP-1 effort were to understand how isotopic and elemental signatures behave during mining, milling, and concentration and to identify analytes that might preserve geologic signatures of the protolith ores. The impurities that are preserved through the concentration process could provide useful forensic signatures and perhaps prove diagnostic of sample origin.

  14. Concentration determination of nucleic acids and proteins using the micro-volume BioSpec-nano-spectrophotometer.

    Science.gov (United States)

    Sukumaran, Suja

    2011-02-17

    Nucleic acid quantitation procedures have advanced significantly in the last three decades. More and more, molecular biologists require consistent small-volume analysis of nucleic acid samples for their experiments. The BioSpec-nano provides a potential solution to the problems of inaccurate, non-reproducible results, inherent in current DNA quantitation methods, via specialized optics and a sensitive PDA detector. The BioSpec-nano also has automated functionality such that mounting, measurement, and cleaning are done by the instrument, thereby eliminating tedious, repetitive, and inconsistent placement of the fiber optic element and manual cleaning. In this study, data is presented on the quantification of DNA and protein, as well as on measurement reproducibility and accuracy. Automated sample contact and rapid scanning allows measurement in three seconds, resulting in excellent throughput. Data analysis is carried out using the built-in features of the software. The formula used for calculating DNA concentration is: Sample Concentration = DF · (OD260-OD320)· NACF (1) Where DF = sample dilution factor and NACF = nucleic acid concentration factor. The Nucleic Acid concentration factor is set in accordance with the analyte selected. Protein concentration results can be expressed as μg/mL or as moles/L by entering e280 and molecular weight values respectively. When residue values for Tyr, Trp and Cysteine (S-S bond) are entered in the e280Calc tab, the extinction coefficient values are calculated as e280 = 5500 x (Trp residues) + 1490 x (Tyr residues) + 125 x (cysteine S-S bond). The e280 value is used by the software for concentration calculation. In addition to concentration determination of nucleic acids and protein, the BioSpec-nano can be used as an ultra micro-volume spectrophotometer for many other analytes or as a standard spectrophotometer using 5 mm pathlength cells.

  15. Concentration Determination of Nucleic Acids and Proteins Using the Micro-volume Bio-spec Nano Spectrophotometer

    Science.gov (United States)

    Sukumaran, Suja

    2011-01-01

    Nucleic Acid quantitation procedures have advanced significantly in the last three decades. More and more, molecular biologists require consistent small-volume analysis of nucleic acid samples for their experiments. The BioSpec-nano provides a potential solution to the problems of inaccurate, non-reproducible results, inherent in current DNA quantitation methods, via specialized optics and a sensitive PDA detector. The BioSpec-nano also has automated functionality such that mounting, measurement, and cleaning are done by the instrument, thereby eliminating tedious, repetitive, and inconsistent placement of the fiber optic element and manual cleaning. In this study, data is presented on the quantification of DNA and protein, as well as on measurement reproducibility and accuracy. Automated sample contact and rapid scanning allows measurement in three seconds, resulting in excellent throughput. Data analysis is carried out using the built-in features of the software. The formula used for calculating DNA concentration is: Sample Concentration = DF · (OD260-OD320)· NACF (1) Where DF = sample dilution factor and NACF = nucleic acid concentration factor. The Nucleic Acid concentration factor is set in accordance with the analyte selected1. Protein concentration results can be expressed as μg/ mL or as moles/L by entering e280 and molecular weight values respectively. When residue values for Tyr, Trp and Cysteine (S-S bond) are entered in the e280Calc tab, the extinction coefficient values are calculated as e280 = 5500 x (Trp residues) + 1490 x (Tyr residues) + 125 x (cysteine S-S bond). The e280 value is used by the software for concentration calculation. In addition to concentration determination of nucleic acids and protein, the BioSpec-nano can be used as an ultra micro-volume spectrophotometer for many other analytes or as a standard spectrophotometer using 5 mm pathlength cells. PMID:21372788

  16. Signaling Pathways Related to Protein Synthesis and Amino Acid Concentration in Pig Skeletal Muscles Depend on the Dietary Protein Level, Genotype and Developmental Stages.

    Directory of Open Access Journals (Sweden)

    Yingying Liu

    Full Text Available Muscle growth is regulated by the homeostatic balance of the biosynthesis and degradation of muscle proteins. To elucidate the molecular interactions among diet, pig genotype, and physiological stage, we examined the effect of dietary protein concentration, pig genotype, and physiological stages on amino acid (AA pools, protein deposition, and related signaling pathways in different types of skeletal muscles. The study used 48 Landrace pigs and 48 pure-bred Bama mini-pigs assigned to each of 2 dietary treatments: lower/GB (Chinese conventional diet- or higher/NRC (National Research Council-protein diet. Diets were fed from 5 weeks of age to respective market weights of each genotype. Samples of biceps femoris muscle (BFM, type I and longissimus dorsi muscle (LDM, type II were collected at nursery, growing, and finishing phases according to the physiological stage of each genotype, to determine the AA concentrations, mRNA levels for growth-related genes in muscles, and protein abundances of mechanistic target of rapamycin (mTOR signaling pathway. Our data showed that the concentrations of most AAs in LDM and BFM of pigs increased (P<0.05 gradually with increasing age. Bama mini-pigs had generally higher (P<0.05 muscle concentrations of flavor-related AA, including Met, Phe, Tyr, Pro, and Ser, compared with Landrace pigs. The mRNA levels for myogenic determining factor, myogenin, myocyte-specific enhancer binding factor 2 A, and myostatin of Bama mini-pigs were higher (P<0.05 than those of Landrace pigs, while total and phosphorylated protein levels for protein kinase B, mTOR, and p70 ribosomal protein S6 kinases (p70S6K, and ratios of p-mTOR/mTOR, p-AKT/AKT, and p-p70S6K/p70S6K were lower (P<0.05. There was a significant pig genotype-dependent effect of dietary protein on the levels for mTOR and p70S6K. When compared with the higher protein-NRC diet, the lower protein-GB diet increased (P<0.05 the levels for mTOR and p70S6K in Bama mini-pigs, but

  17. Effects of Long-Term Storage Time and Original Sampling Month on Biobank Plasma Protein Concentrations

    Directory of Open Access Journals (Sweden)

    Stefan Enroth

    2016-10-01

    Full Text Available The quality of clinical biobank samples is crucial to their value for life sciences research. A number of factors related to the collection and storage of samples may affect the biomolecular composition. We have studied the effect of long-time freezer storage, chronological age at sampling, season and month of the year and on the abundance levels of 108 proteins in 380 plasma samples collected from 106 Swedish women. Storage time affected 18 proteins and explained 4.8–34.9% of the observed variance. Chronological age at sample collection after adjustment for storage-time affected 70 proteins and explained 1.1–33.5% of the variance. Seasonal variation had an effect on 15 proteins and month (number of sun hours affected 36 proteins and explained up to 4.5% of the variance after adjustment for storage-time and age. The results show that freezer storage time and collection date (month and season exerted similar effect sizes as age on the protein abundance levels. This implies that information on the sample handling history, in particular storage time, should be regarded as equally prominent covariates as age or gender and need to be included in epidemiological studies involving protein levels.

  18. Rising atmospheric CO2 is reducing the protein concentration of a floral pollen source essential for North American bees.

    Science.gov (United States)

    Ziska, Lewis H; Pettis, Jeffery S; Edwards, Joan; Hancock, Jillian E; Tomecek, Martha B; Clark, Andrew; Dukes, Jeffrey S; Loladze, Irakli; Polley, H Wayne

    2016-04-13

    At present, there is substantive evidence that the nutritional content of agriculturally important food crops will decrease in response to rising levels of atmospheric carbon dioxide, Ca However, whether Ca-induced declines in nutritional quality are also occurring for pollinator food sources is unknown. Flowering late in the season, goldenrod (Solidago spp.) pollen is a widely available autumnal food source commonly acknowledged by apiarists to be essential to native bee (e.g. Bombus spp.) and honeybee (Apis mellifera) health and winter survival. Using floral collections obtained from the Smithsonian Natural History Museum, we quantified Ca-induced temporal changes in pollen protein concentration of Canada goldenrod (Solidago canadensis), the most wide spread Solidago taxon, from hundreds of samples collected throughout the USA and southern Canada over the period 1842-2014 (i.e. a Ca from approx. 280 to 398 ppm). In addition, we conducted a 2 year in situtrial of S. Canadensis populations grown along a continuous Ca gradient from approximately 280 to 500 ppm. The historical data indicated a strong significant correlation between recent increases in Ca and reductions in pollen protein concentration (r(2)= 0.81). Experimental data confirmed this decrease in pollen protein concentration, and indicated that it would be ongoing as Ca continues to rise in the near term, i.e. to 500 ppm (r(2)= 0.88). While additional data are needed to quantify the subsequent effects of reduced protein concentration for Canada goldenrod on bee health and population stability, these results are the first to indicate that increasing Ca can reduce protein content of a floral pollen source widely used by North American bees. © 2016 The Author(s).

  19. Age- and gender-related alteration in plasma advanced oxidation protein products (AOPP) and glycosaminoglycan (GAG) concentrations in physiological ageing.

    Science.gov (United States)

    Komosinska-Vassev, Katarzyna; Olczyk, Pawel; Winsz-Szczotka, Katarzyna; Kuznik-Trocha, Kornelia; Klimek, Katarzyna; Olczyk, Krystyna

    2012-02-13

    The authors studied the role of increased oxidative stress in the development of oxidative protein damage and extracellular matrix (ECM) components in ageing. The age- and gender-associated disturbances in connective tissue metabolism were evaluated by the plasma chondroitin sulphated glycosaminoglycans (CS-GAG) and non-sulphated GAG-hyaluronan (HA) measurements. Plasma concentration of advanced oxidation protein products (AOPP) was analysed in order to assess oxidative protein damage and evaluate the possible deleterious role of oxidative phenomenon on tissue proteoglycans' metabolism during the physiological ageing process. Sulphated and non-sulphated GAGs as well as AOPP were quantified in plasma samples from 177 healthy volunteers. A linear age-related decline of plasma CS-GAG level was found in this study (r=-0.46; page (r=0.44; page-dependent relationship has been shown in regard to AOPP. AOPP levels significantly increased with age (r=0.63; pphysiological ageing. A significant correlation was found between the concentrations of AOPP and both CS-GAG (r=-0.31; page changes in the ECM are reflected by CS-GAG and HA plasma levels. Strong correlations between AOPP and ECM components indicate that oxidative stress targets protein and non-protein components of the connective tissue matrix during human ageing.

  20. Physical and sensory characteristics of pellets elaborated with different levels of corn grits and whey protein concentrate

    Directory of Open Access Journals (Sweden)

    Anderson Felicori Fernandes

    2016-04-01

    Full Text Available ABSTRACT Whey has proteins of high biological value, which has been used as an ingredient in the elaboration of yogurt, milk beverages and as protein concentrates. Food extrusion stands out as one of the most efficient cooking techniques, allowing a number of product types, from soluble flour to convenience products, such as snacks, which have high acceptability by the consumers. Products processed by extrusion, such as those expanded by frying (pellets, have high carbohydrate content, and its enrichment with protein can favor its nutritional aspect. This study aimed to use the whey protein concentrate (WPC in combination with corn grits in the preparation of pellets. Absolute density, density of expanded pellets, color, crispness index, and pellet sensory acceptance were determined. For the absolute density, contents from 5% to 17% produced denser non-expanded pellets. The higher the WPC content and the temperature of the extruder, the higher the density of the expanded pellets. The crispness index was not altered by the protein content and by the extruder temperature. In the sensory analysis, the preferred samples were the ones with lower WPC levels (5%. We concluded that higher WPC values in the pellets formulation increased their density, but did not alter color and texture, as well as small WPC levels did not affect the acceptance of snacks.

  1. Effects of Protein-Iron Complex Concentrate Supplementation on Iron Metabolism, Oxidative and Immune Status in Preweaning Calves

    Directory of Open Access Journals (Sweden)

    Robert Kupczyński

    2017-07-01

    Full Text Available The objective of this study was to determine the effects of feeding protein-iron complex (PIC on productive performance and indicators of iron metabolism, hematology parameters, antioxidant and immune status during first 35 days of a calf’s life. Preparation of the complex involved enzymatic hydrolysis of milk casein (serine protease from Yarrowia lipolytica yeast. Iron chloride was then added to the hydrolyzate and lyophilizate. Calves were divided into treated groups: LFe (low iron dose 10 g/day calf of protein-iron complex, HFe (height iron dose 20 g/day calf, and control group. Dietary supplements containing the lower dose of concentrate had a significant positive effect on iron metabolism, while the higher dose of concentrate resulted in increase of total iron binding capacity (TIBC, saturation of transferrin and decrease of and unsaturated iron binding capacity (UIBC, which suggest iron overload. Additionally, treatment with the lower dose of iron remarkably increased the antioxidant parameters, mainly total antioxidant (TAS and glutathione peroxidase activity (GPx. Higher doses of PIC were related to lower total antioxidant status. IgG, IgM, insulin, glucose, TNFα and IGF-1 concentration did not change significantly in either group after supplementation. In practice, the use of protein-iron complex concentrate requires taking into account the iron content in milk replacers and other feedstuffs.

  2. Rapid concentration and dialysis of proteins with single hollow fibers: possible applications in analysis of protein secretion by isolated cells and steroid radioimmunoassays

    International Nuclear Information System (INIS)

    Rommerts, F.F.G.; Clotscher, W.F.; Van der Molen, H.J.

    1977-01-01

    Single hollow fibers were used in specially made cells for fast concentration and dialysis of solutions containing macromolecules. Volumes on the order of 5 ml of diluted protein solutions could be concentrated to 50--100 μl or less within 7 min with a protein recovery of 60--80%. More than 99% of the molecules with a molecular weight less than 500 could be removed in less than 1 hr. A possible application of the rapid dialysis method for the mechanization of radioimmunoassays is indicated. It was shown that in the radioimmunoassay of steriods the unbound steroids could be removed after incubation with antiserum, within 10 min and without a change in volume

  3. The Effect of Serum 25-Hydroxyvitamin D Concentrations on Elevated Serum C-Reactive Protein Concentrations in Normal Weight, Overweight and Obese Participants of a Preventive Health Program.

    Science.gov (United States)

    Mastroeni, Silmara S B S; Munasinghe, Lalani L; Pham, Truong-Minh; Loehr, Sarah A; Ekwaru, John Paul; Mastroeni, Marco F; Veugelers, Paul J

    2016-11-04

    The hypothesized effect of vitamin D on C-reactive protein (CRP) has received substantial attention as a potential means to alleviate the risk for cardiovascular disease. However, observational studies have been inconsistent in their reporting of associations between serum 25-hydroxyvitamin D (25(OH)D) and CRP concentrations, and trials and meta analyses have been inconsistent in their conclusions regarding the effect of vitamin D supplementation on CRP concentrations. These supplementation trials were mostly conducted among patients with more or less inflammatory complications and did not consider potential distinctive effects by weight status. To further our understanding of the potential influences of vitamin D on CRP, we analyzed longitudinal observations of 6755 participants of a preventative health program. On average, serum 25(OH)D concentrations increased from 88.3 to 121.0 nmol/L and those of CRP decreased from 1.7 to 1.6 mg/L between baseline and follow up. Relative to obese participants without temporal increases in 25(OH)D, those who showed improvements of <25, 25-50, 50-75, and more than 75 nmol/L at follow up were 0.57 (95% confidence interval: 0.37-0.88), 0.54 (0.34-0.85), 0.49 (0.30-0.80), and 0.48 (0.29-0.78) times as likely to have elevated CRP concentrations (≥1 mg/L), respectively. These associations were less pronounced and not statistically significant for normal weight and overweight participants. Herewith, the findings suggest that promotion of adequate serum 25(OH)D concentrations among obese individuals along with healthy lifestyles may alleviate the public health burden associated with cardiovascular disease.

  4. Expression, purification, crystallization and preliminary X-ray analysis of eCGP123, an extremely stable monomeric green fluorescent protein with reversible photoswitching properties

    International Nuclear Information System (INIS)

    Don Paul, Craig; Traore, Daouda A. K.; Byres, Emma; Rossjohn, Jamie; Devenish, Rodney J.; Kiss, Csaba; Bradbury, Andrew; Wilce, Matthew C. J.; Prescott, Mark

    2011-01-01

    eCGP123, an extremely stable GFP with photoswitching properties, has been expressed, purified and crystallized. A diffraction data set has been collected at 2.10 Å resolution. Enhanced consensus green protein variant 123 (eCGP123) is an extremely thermostable green fluorescent protein (GFP) that exhibits useful negative reversible photoswitching properties. eCGP123 was derived by the application of both a consensus engineering approach and a recursive evolutionary process. Diffraction-quality crystals of recombinant eCGP123 were obtained by the hanging-drop vapour-diffusion method using PEG 3350 as the precipitant. The eCGP123 crystal diffracted X-rays to 2.10 Å resolution. The data were indexed in space group P1, with unit-cell parameters a = 74.63, b = 75.38, c = 84.51 Å, α = 90.96, β = 89.92, γ = 104.03°. The Matthews coefficient (V M = 2.26 Å 3 Da −1 ) and a solvent content of 46% indicated that the asymmetric unit contained eight eCGP123 molecules

  5. Dietary factor VII activation does not increase plasma concentrations of prothrombin fragment 1+2 in patients with stable angina pectoris and coronary atherosclerosis

    DEFF Research Database (Denmark)

    Bladbjerg, E-M; Münster, A M; Marckmann, P

    2000-01-01

    Studies in healthy subjects showed that blood coagulation factor VII (FVII) is activated postprandially after consumption of high-fat meals, but accompanying thrombin formation has not been demonstrated. In patients with coronary atherosclerosis, the arterial intima is supposed to present more...... tissue factor, the cofactor of FVII, to circulating blood; therefore, thrombin formation in response to FVII activation is more likely to occur in such patients. This hypothesis was tested in a randomized crossover study of 30 patients (aged 43 to 70 years) with stable angina pectoris...

  6. Preparation of Low Allergenic Protein Concentrated Natural Rubber Latex Using Suitable Low Molecular Weight Cellulose Derivatives Induced by Gamma Irradiation

    International Nuclear Information System (INIS)

    Siri-Upathum, Chyagrit; Boonyawat, Jariya

    2007-08-01

    Full text: Low molecular weight carboxy methyl cellulose (CMC), hydroxyl ethyl cellulose (HEC), hydroxyl propyl cellulose (HPC) and methyl cellulose (MC) prepared by radiation-induced degradation were added into diluted natural concentrated latex prior to centrifuge for a purpose of reducing allergenic rubber protein in the latex. Optimum molecular weight (Mv) of CMC and HEC for such a purpose was found to be 17-18 kDa which decreased allergenic rubber protein (14-94 kDa) to an undetectable amount as determined by SDS PAGE method

  7. Effects of dietary incorporation of potato protein concentrate and supplementation of methionine on growth and feed utilization of rainbow trout

    DEFF Research Database (Denmark)

    Xie, S.; Jokumsen, Alfred

    1998-01-01

    and supplementation of methionine in the diet of rainbow trout. When the proportion of PPC exceeded 56 g kg-1 the growth of fish decreased while both growth and feed utilization decreased when the dietary PPC was 111 g kg-1. Protein productive value and condition factor of the fish decreased and mortality increased......Four diets (1, 2, 3 and 4) were formulated to contain different potato protein concentrate (PPC) levels (0, 22, 56, and 111 g kg-1). Diet 5 contained 56 g kg-1 PPC and 17 g kg-1 methionine. A growth trial was conducted to investigate the effect on growth and feed utilization of incorporation of PPC...

  8. Matrix Gla Protein polymorphism, but not concentrations, is associated with radiographic hand osteoarthritis

    Science.gov (United States)

    Objective. Factors associated with mineralization and osteophyte formation in osteoarthritis (OA) are incompletely understood. Genetic polymorphisms of matrix Gla protein (MGP), a mineralization inhibitor, have been associated clinically with conditions of abnormal calcification. We therefore evalua...

  9. Method for determining the concentration of adsorbed protein and cell biomass in cellulose fermentations

    Energy Technology Data Exchange (ETDEWEB)

    Moreira, A R; Phillips, J A; Humphrey, A E

    1978-09-01

    The method presented is based on the determination of the total Lowry protein of the solids and the total Kjeldahl nitrogen of the solids. Experimental data proving the validity of the method are reported. (JSR)

  10. Can serum concentration of C-reactive protein, albumin and body ...

    African Journals Online (AJOL)

    ... of acute phase proteins and a fall in CD4+T-cell counts and body weight. ... subjects (control) attending the HIV/AIDS Clinic in Nnamdi Azikiwe Teaching Hospital, ... while their weights were measured using high precision weighing balance.

  11. Effects of egg white protein supplementation on muscle strength and serum free amino acid concentrations.

    Science.gov (United States)

    Hida, Azumi; Hasegawa, Yuko; Mekata, Yuko; Usuda, Mika; Masuda, Yasunobu; Kawano, Hitoshi; Kawano, Yukari

    2012-10-19

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

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

    Directory of Open Access Journals (Sweden)

    Yukari Kawano

    2012-10-01

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

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

    Science.gov (United States)

    Hida, Azumi; Hasegawa, Yuko; Mekata, Yuko; Usuda, Mika; Masuda, Yasunobu; Kawano, Hitoshi; Kawano, Yukari

    2012-01-01

    The aim of this study was to evaluate the effects of egg white protein compared to carbohydrate intake prior to exercise on fat free mass (FFM), one repetition maximum (1RM) muscle strength and blood biochemistry in female athletes. Thirty healthy female collegiate athletes were recruited for this study and matched by sport type, body fat percentage and 1RM leg curl muscle strength. Participants were randomly divided into two groups: protein group (15.0 g egg white protein; 75 kcal) and carbohydrate group (17.5 g maltodextrin, 78 kcal). Supplements were administered daily at the same time in a double-blind manner prior to training during an 8-week period. Measurements were performed before and after the 8-week regimen. The mean dietary energy intake did not change throughout the study period. FFM and 1RM assessments (i.e., leg curl, leg extension, squat, and bench press) increased in both groups. Furthermore, serum urea and serum citrulline levels after the 8-week regimen increased significantly only in the protein group. Our findings indicated that compared to the carbohydrate supplement, the protein supplement was associated with some changes in protein metabolites but not with changes in body composition or muscle strength. PMID:23201768

  14. Plasma protein concentration and control of coronary vascular resistance in isolated rat heart

    NARCIS (Netherlands)

    Avolio, A. P.; Spaan, J. A.; Laird, J. D.

    1980-01-01

    Isolated externally paced (300 beats/min) rat hearts were perfused at constant pressure (70 mmHg) using a modified Krebs-Henseleit solution with (n = 52) and without (n = 15) washed bovine red cells. Albumin concentration varied from 1 to 10 g/dl. With increasing albumin concentration in

  15. Investigation of mercury-containing proteins by enriched stable isotopic tracer and size-exclusion chromatography hyphenated to inductively coupled plasma-isotope dilution mass spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Shi Junwen [Laboratory for Bio-Environmental Health Sciences of Nanoscale Materials and Key Laboratory of Nuclear Analytical Techniques, Institute of High Energy Physics, Chinese Academy of Sciences, Beijing 100049 (China)]|[Graduate School of the Chinese Academy of Sciences, Beijing 100049 (China); Feng Weiyue [Laboratory for Bio-Environmental Health Sciences of Nanoscale Materials and Key Laboratory of Nuclear Analytical Techniques, Institute of High Energy Physics, Chinese Academy of Sciences, Beijing 100049 (China)]. E-mail: fengwy@mail.ihep.ac.cn; Wang Meng [Laboratory for Bio-Environmental Health Sciences of Nanoscale Materials and Key Laboratory of Nuclear Analytical Techniques, Institute of High Energy Physics, Chinese Academy of Sciences, Beijing 100049 (China)]|[Graduate School of the Chinese Academy of Sciences, Beijing 100049 (China); Zhang Fang [Graduate School of the Chinese Academy of Sciences, Beijing 100049 (China); Li Bai [Laboratory for Bio-Environmental Health Sciences of Nanoscale Materials and Key Laboratory of Nuclear Analytical Techniques, Institute of High Energy Physics, Chinese Academy of Sciences, Beijing 100049 (China); Wang Bing; Zhu Motao [Laboratory for Bio-Environmental Health Sciences of Nanoscale Materials and Key Laboratory of Nuclear Analytical Techniques, Institute of High Energy Physics, Chinese Academy of Sciences, Beijing 100049 (China)]|[Graduate School of the Chinese Academy of Sciences, Beijing 100049 (China); Chai Zhifang [Laboratory for Bio-Environmental Health Sciences of Nanoscale Materials and Key Laboratory of Nuclear Analytical Techniques, Institute of High Energy Physics, Chinese Academy of Sciences, Beijing 100049 (China)]|[Institute of Nuclear Technology, Shenzhen University, Shenzhen 518060 (China)]|[Institute of Nanochemistry and Nanosafety, Shanghai University, Shanghai (China)

    2007-01-30

    In order to investigate trace mercury-containing proteins in maternal rat and their offspring, a method of enriched stable isotopic tracer ({sup 196}Hg and {sup 198}Hg) combined with size-exclusion chromatography (SEC) coupled to inductively coupled plasma-isotope dilution mass spectrometry (ICP-IDMS) was developed. Prior to the analysis, {sup 196}Hg- and {sup 198}Hg-enriched methylmercury was administrated to the pregnant rats. Then the mercury-containing proteins in serum and brain cytosol of the dam and pup rats were separated by size-exclusion columns and the mercury was detected by ICP-MS. The ICP-MS spectrogram of the tracing samples showed significantly elevated {sup 196}Hg and {sup 198}Hg isotopic signals compared with the natural ones, indicating that the detection sensitivity could be increased by the tracer method. The contents of mercury in chromatographic fractions of the dam and pup rat brain cytosol were quantitatively estimated by post-column reverse ID-ICP-MS. The quantitative speciation differences of mercury in brain cytosol between the dam and pup rats were observed, indicating that such studies could be useful for toxicological estimation. Additionally, the isotopic ratio measurement of {sup 198}Hg/{sup 202}Hg in the tracing samples could be used to identify the artifact mercury species caused in the analytical procedure. The study demonstrates that the tracer method combined with high-performance liquid chromatography (HPLC)-ICP-IDMS could provide reliably qualitative and quantitative information on mercury-containing proteins in organisms.

  16. Tracing metabolic routes of dietary carbohydrate and protein in rainbow trout (Oncorhynchus mykiss) using stable isotopes ([¹³C]starch and [¹⁵N]protein): effects of gelatinisation of starches and sustained swimming.

    Science.gov (United States)

    Felip, Olga; Ibarz, Antoni; Fernández-Borràs, Jaume; Beltrán, Marta; Martín-Pérez, Miguel; Planas, Josep V; Blasco, Josefina

    2012-03-01

    Here we examined the use of stable isotopes, [¹³C]starch and [¹⁵N]protein, as dietary tracers to study carbohydrate assimilation and distribution and protein utilisation, respectively, by rainbow trout (Oncorhynchus mykiss). The capacity of glucose uptake and use by tissues was studied, first, by varying the digestibility of carbohydrate-rich diets (30 % carbohydrate), using raw starch and gelatinised starch (GS) and, second, by observing the effects of two regimens of activity (voluntary swimming, control; sustained swimming at 1·3 body lengths/s, exercise) on the GS diet. Isotopic ratio enrichment (¹³C and ¹⁵N) of the various tissue components (protein, lipid and glycogen) was measured in the liver, muscles, viscera and the rest of the fish at 11 and 24 h after a forced meal. A level of 30 % of digestible carbohydrates in the food exceeded the capacity of rainbow trout to use this nutrient, causing long-lasting hyperglycaemia that raises glucose uptake by tissues, and the synthesis of glycogen and lipid in liver. Total 13C recovered 24 h post-feeding in the GS group was lower than at 11 h, indicating a proportional increase in glucose oxidation, although the deposition of lipids in white muscle (WM) increased. Prolonged hyperglycaemia was prevented by exercise, since sustained swimming enhances the use of dietary carbohydrates, mainly through conversion to lipids in liver and oxidation in muscles, especially in red muscle (RM). Higher recoveries of total 15N for exercised fish at 24 h, mainly into the protein fraction of both RM and WM, provide evidence that sustained swimming improves protein deposition, resulting in an enhancement of the protein-sparing effect.

  17. Serum zinc, copper, retinol-binding protein, prealbumin, and ceruloplasmin concentrations in infants receiving intravenous zinc and copper supplementation.

    Science.gov (United States)

    Lockitch, G; Godolphin, W; Pendray, M R; Riddell, D; Quigley, G

    1983-02-01

    One hundred twenty-seven newborn infants requiring parenteral nutrition were randomly assigned to receive differing amounts of zinc (40 to 400 micrograms/kg/day) and copper (20 or 40 micrograms/kg/day) supplementation within five birth weight groups (600 to 2,500 gm). The serum zinc concentration remained relatively constant in the group receiving the most zinc supplementation after two weeks of therapy, but declined sharply in the groups receiving less supplementation. No effect of increased copper intake was noted on ceruloplasmin values, but a difference in serum copper concentrations was noted at two weeks. No correlation was noted between serum zinc and copper values or among those for serum zinc, retinol-binding protein, and prealbumin. Reference ranges were defined for serum zinc, copper, retinol-binding protein, prealbumin, and ceruloplasmin in the preterm infant.

  18. Serum C-reactive protein concentration as an indicator of remission status in dogs with multicentric lymphoma

    DEFF Research Database (Denmark)

    Nielsen, Lise; Toft, Nils; Eckersall, David

    2007-01-01

    Background: The acute-phase protein C-reactive protein (CRP) is used as a diagnostic and prognostic marker in humans with various neoplasias, including non-Hodgkin's lymphoma. Objective: To evaluate if CRP could be used to detect different remission states in dogs with lymphoma. Animals: Twenty......-two dogs with untreated multicentric lymphoma. Methods: Prospective observational study. Blood samples were collected at the time of diagnosis, before each chemotherapy session, and at follow-up visits, resulting in 287 serum samples. Results: Before therapy, a statistically significant majority...... of the dogs (P = .0019) had CRP concentrations above the reference range (68%, 15/22). After achieving complete remission 90% (18/20) of the dogs had CRP concentrations within the reference range, and the difference in values before and after treatment was statistically significant (P

  19. The association between economic development, lifestyle differentiation, and C-reactive protein concentration within rural communities in Hainan Island, China.

    Science.gov (United States)

    Inoue, Yosuke; Stickley, Andrew; Yazawa, Aki; Li, Dandan; Du, Jianwei; Jin, Yuming; Chen, Yan; Watanabe, Chiho

    2016-01-01

    Earlier fieldwork in rural areas of Hainan Island, China, demonstrated that during the course of economic development increasing differences had emerged in lifestyles within communities. It is possible that these variations might have stratified residents into subpopulations with different health attributes. This study examined the association between C-reactive protein (CRP) concentration, a biomarker of future cardiovascular events, and personal lifestyle parameters and the degree of community-level economic development among rural communities. A cross-sectional field survey was undertaken in 19 rural communities in Hainan. Convenience sampling was used to recruit 1,744 participants. Dried blood spot samples were collected to measure high-sensitivity CRP concentration. Sex-stratified multilevel regression analyses were conducted to identify factors associated with CRP concentration among the participants. While CRP concentration was negatively associated with being married and (more) education among men, for women CRP concentration was associated with the frequency of poultry consumption (P = 0.014) and the experience of migratory work in the previous year (P = 0.009). In addition, for females, living in communities with a greater degree of inequality, as indexed by the Gini coefficient, was also associated with increased CRP concentration (P = 0.003). Given that CRP concentration is a marker of future CVD risk, this study suggests that within these previously homogenous rural communities, economic development might have stratified people into population subgroups with a different CVD risk. © 2015 Wiley Periodicals, Inc.

  20. Concentration of total protein and degree of acidity (pH of saliva when fasting and after breakfasting

    Directory of Open Access Journals (Sweden)

    Gemella Nur Illahi

    2016-04-01

    Full Text Available Background: While fasting, the mouth does not work to eat and drink so that the salivary glands become less active so saliva production decreased and there was a change in eating timewhich is relation to the mastication process that impact on changes in the degree of acidity (pH Objectives: To determine the concentration of total protein and the degree of acidity (pH of saliva when fasting and after breakfasting. Materials and Methods: The study was observational analytic design with longitudinal (follow up study conducted in the Hj. Halima Dg. Sikati Dental Hospital inKandea in July 2015, the sampling method was purposive sampling. Population was 35 clinical students at the Department of Dental Public Health, Faculty of Dentistry Hasanuddin University with a total sample of 16 students who fit the criteria of the study subjects. To calculate the total protein of saliva concentration using Kyltecautoanalyzerand pH meter to measure the acidity of saliva. Data was analyzed was using SPSS version 17.0 (paired t-test, p <0.05. Results: The mean of total protein (% while fasting by 0135% ± 0.026 and the mean total protein (% after breakfasting at 0.179% ± 0.035, while the average degree of acidity (pH during fasting at 7.26 ± 0:24 and the average degree of acidity (pH after breakfasting at 7.66 ± 0.23 with p-value (0.000. Conclusions: An increase in the total protein concentration and acidity (pH after breakfasting.

  1. Physiochemical properties, microstructure, and probiotic survivability of nonfat goats' milk yogurt using heat-treated whey protein concentrate as fat replacer.

    Science.gov (United States)

    Zhang, Tiehua; McCarthy, James; Wang, Guorong; Liu, Yanyan; Guo, Mingruo

    2015-04-01

    There is a market demand for nonfat fermented goats' milk products. A nonfat goats' milk yogurt containing probiotics (Lactobacillus acidophilus, and Bifidobacterium spp.) was developed using heat-treated whey protein concentrate (HWPC) as a fat replacer and pectin as a thickening agent. Yogurts containing untreated whey protein concentrate (WPC) and pectin, and the one with only pectin were also prepared. Skim cows' milk yogurt with pectin was also made as a control. The yogurts were analyzed for chemical composition, water holding capacity (syneresis), microstructure, changes in pH and viscosity, mold, yeast and coliform counts, and probiotic survivability during storage at 4 °C for 10 wk. The results showed that the nonfat goats' milk yogurt made with 1.2% HWPC (WPC solution heated at 85 °C for 30 min at pH 8.5) and 0.35% pectin had significantly higher viscosity (P yogurts and lower syneresis than the goats' yogurt with only pectin (P yogurt samples did not change much throughout storage. Bifidobacterium spp. remained stable and was above 10(6) CFU g(-1) during the 10-wk storage. However, the population of Lactobacillus acidophilus dropped to below 10(6) CFU g(-1) after 2 wk of storage. Microstructure analysis of the nonfat goats' milk yogurt by scanning electron microscopy revealed that HWPC interacted with casein micelles to form a relatively compact network in the yogurt gel. The results indicated that HWPC could be used as a fat replacer for improving the consistency of nonfat goats' milk yogurt and other similar products. © 2015 Institute of Food Technologists®

  2. Effects of Boundary Layer Height on the Model of Ground-Level PM2.5 Concentrations from AOD: Comparison of Stable and Convective Boundary Layer Heights from Different Methods

    Directory of Open Access Journals (Sweden)

    Zengliang Zang

    2017-06-01

    Full Text Available The aerosol optical depth (AOD from satellites or ground-based sun photometer spectral observations has been widely used to estimate ground-level PM2.5 concentrations by regression methods. The boundary layer height (BLH is a popular factor in the regression model of AOD and PM2.5, but its effect is often uncertain. This may result from the structures between the stable and convective BLHs and from the calculation methods of the BLH. In this study, the boundary layer is divided into two types of stable and convective boundary layer, and the BLH is calculated using different methods from radiosonde data and National Centers for Environmental Prediction (NCEP reanalysis data for the station in Beijing, China during 2014–2015. The BLH values from these methods show significant differences for both the stable and convective boundary layer. Then, these BLHs were introduced into the regression model of AOD-PM2.5 to seek the respective optimal BLH for the two types of boundary layer. It was found that the optimal BLH for the stable boundary layer is determined using the method of surface-based inversion, and the optimal BLH for the convective layer is determined using the method of elevated inversion. Finally, the optimal BLH and other meteorological parameters were combined to predict the PM2.5 concentrations using the stepwise regression method. The results indicate that for the stable boundary layer, the optimal stepwise regression model includes the factors of surface relative humidity, BLH, and surface temperature. These three factors can significantly enhance the prediction accuracy of ground-level PM2.5 concentrations, with an increase of determination coefficient from 0.50 to 0.68. For the convective boundary layer, however, the optimal stepwise regression model includes the factors of BLH and surface wind speed. These two factors improve the determination coefficient, with a relatively low increase from 0.65 to 0.70. It is found that the

  3. Effect of prolonged exposure to sublethal concentrations of DDT and DDE on protein expression in human pancreatic beta cells.

    Science.gov (United States)

    Pavlikova, Nela; Smetana, Pavel; Halada, Petr; Kovar, Jan

    2015-10-01

    Pollution of the environment represents one of less explored potential reasons for the worldwide epidemic of type 2 diabetes. One of the most prevalent organochlorine pollutants remains the pesticide DDT and its degradation product DDE. Despite some epidemiologic correlations between levels of DDT and DDE in human organism and the prevalence of diabetes, there is almost no information about the exact targets of these compounds inside pancreatic beta cells. To detect functional areas of pancreatic beta cells that could be affected by exposure to DDT and DDE, we analyzed changes in protein expression in the NES2Y human pancreatic beta cell line exposed to three sublethal concentrations (0.1 μM, 1 μM, 10 μM) of DDT and DDE for 1 month. Protein separation and identification was achieved using high-resolution 2D-electrophoresis, computer analysis and mass spectrometry. With these techniques, four proteins were found downregulated after exposure to 10 μM DDT: three cytoskeletal proteins (cytokeratin 8, cytokeratin 18 and actin) and one protein involved in glycolysis (alpha-enolase). Two proteins were downregulated after exposure to 10 μM DDE: cytokeratin 18 and heterogenous nuclear ribonucleoprotein H1 (HNRH1). These changes correlate with previously described effects of other stress conditions (e.g. exposure to palmitate, hyperglycemia, imidazoline derivative, and cytokines) on protein expression in pancreatic beta cells. We conclude that cytoskeletal proteins and their processing, glucose metabolism, and mRNA processing may represent targets affected by exposure to conditions hostile to pancreatic beta cells, including exposure to DDT and DDE. Copyright © 2015 Elsevier Inc. All rights reserved.

  4. Identification and changes in the seasonal concentrations of heat shock proteins in roe deer (Capreolus capreolus) epididymides.

    Science.gov (United States)

    Majewska, A M; Kordan, W; Koziorowska-Gilun, M; Wysocki, P

    2017-02-01

    Heat shock proteins (HSPs) act as molecular chaperones with important regulatory functions. HSPs are considered to be essential factors in animal reproduction. In view of seasonal variations in the secretory activity of the reproductive tract of mature roe deer (Capreolus capreolus), the aims of this study were to identify HSPs in the epididymides and compare the expression of the identified proteins in three periods of the reproductive season. Two-dimensional polyacrylamide gel electrophoresis revealed the highest number of polypeptides in homogenates of epididymal tissues and in caput, corpus and cauda epididymal fluids throughout the reproductive season. Epididymal tissue homogenates and epididymal fluids were analysed by tandem mass spectrometry (MS/MS) to reveal 31 polypeptides with enzymatic activity, including polypeptides with antioxidant properties, structural and cell signalling functions. Moreover, among the identified polypeptides, five of them were similar to heat shock proteins: endoplasmin (Grp94); heat shock protein 90 kDa (HSP90); 78-kDa glucose-regulated protein (Grp78); chain A, the crystal structure of the human HSP70 ATPase domain and heat shock protein beta-1 isoform X. The concentrations of the analysed polypeptides, expressed in optical density units (ODU), differed significantly (p ≤ .05) across the examined periods of the reproductive season. The highest ODU values for almost all analysed proteins were observed during the rutting period. The presence of HSPs in the epididymal tissues and fluids of roe deer in different periods of the reproductive season could indicate that those proteins play an important role in sperm maturation in the epididymis. © 2016 Blackwell Verlag GmbH.

  5. Hydrolysis with Cucurbita ficifolia serine protease reduces antigenic response to bovine whey protein concentrate and αs-casein.

    Science.gov (United States)

    Babij, Konrad; Bajzert, Joanna; Dąbrowska, Anna; Szołtysik, Marek; Zambrowicz, Aleksandra; Lubec, Gert; Stefaniak, Tadeusz; Willak-Janc, Ewa; Chrzanowska, Józefa

    2015-11-01

    In the present study the effect of hydrolysis with non-commercial Cucurbita ficifolia serine protease on a reduction of the IgE and IgG binding capacity of whey protein concentrate and αs-casein was investigated. The intensity of the protein degradation was analyzed by the degree of hydrolysis, the free amino groups content and RP-HPLC. The ability to bind the antibodies by native proteins and their hydrolysates was determined using a competitive ELISA test. Deep hydrolysis contributed to a significant reduction of immunoreactive epitopes present in WPC. In the case of IgE and IgG present in the serum pool of children with CMA, the lowest binding capacity was detected in the 24 h WPC hydrolysate, where the inhibition of the reaction with native WPC was ≤23 and ≤60 %, respectively. The analysis of the IgG reactivity in the antiserum of the immunized goat showed that the lowest antibody binding capacity was exhibited also by 24 h WPC hydrolysate at a concentration of 1000 μg/ml where the inhibition of the reaction with nWPC was ≤47 %. One-hour hydrolysis of α-casein was sufficient to significant reduction of the protein antigenicity, while the longer time (5 h) of hydrolysis probably lead to the appearance of new epitopes reactive with polyclonal.

  6. Influence of Bovine Whey Protein Concentrate and Hydrolysate Preparation Methods on Motility in the Isolated Rat Distal Colon

    Science.gov (United States)

    Dalziel, Julie E.; Anderson, Rachel C.; Bassett, Shalome A.; Lloyd-West, Catherine M.; Haggarty, Neill W.; Roy, Nicole C.

    2016-01-01

    Whey protein concentrate (WPC) and hydrolysate (WPH) are protein ingredients used in sports, medical and pediatric formulations. Concentration and hydrolysis methods vary for whey sourced from cheese and casein co-products. The purpose of this research was to investigate the influence of whey processing methods on in vitro gastrointestinal (GI) health indicators for colonic motility, epithelial barrier integrity and immune modulation. WPCs from casein or cheese processing and WPH (11% or 19% degree of hydrolysis, DH) were compared for their effects on motility in a 1 cm section of isolated rat distal colon in an oxygenated tissue bath. Results showed that WPC decreased motility irrespective of whether it was a by-product of lactic acid or mineral acid casein production, or from cheese production. This indicated that regardless of the preparation methodology, the whey protein contained components that modulate aspects of motility within the distal colon. WPH (11% DH) increased contractile frequency by 27% in a delayed manner and WPH (19% DH) had an immediate effect on contractile properties, increasing tension by 65% and frequency by 131%. Increased motility was associated with increased hydrolysis that may be attributed to the abundance of bioactive peptides. Increased frequency of contractions by WPH (19% DH) was inhibited (by 44%) by naloxone, implicating a potential involvement of opioid receptors in modulation of motility. Trans-epithelial electrical resistance and cytokine expression assays revealed that the WPC proteins studied did not alter intestinal barrier integrity or elicit any discernible immune response. PMID:27983629

  7. Free Fatty Acid Concentration and Carboxy methyl cellulase Activity of Some Formulas of Protected Fat-proteins Tested In Vitro

    Directory of Open Access Journals (Sweden)

    Lilis Hartati

    2015-05-01

    Full Text Available The aim of this study was to determine the levels of free fatty acids and carboxymethylcellulase activity (cmc-ase activity of some protected fat-proteins base on in vitro Tilley and Terry method. Two sources of fat, i.e. crude palm oil and fish oil and three sources of protein i.e. skim milk, soybean flour and soybean meal were used in the formulation of protected fat-protein, and thus there were six treatment combinations. The filtrate from the in vitro test was analyzed for the levels of free fatty acids and  cmcase activity. The result of this research indicates that different combinations of feed materials and fat give different content of free fatty acid in first stage and second stage in vitro, with the best results in the combination treatment of skim milk and palm oil that give the lowest result of  free fatty acid concentration in fisrt stage in vitro (0.168% and the highest result free fatty acid concentration in second stage in vitro ( 4.312% . The activity of CMC-ase was not influenced by different  sources of fat and protein. It can be concluded was that the protection of the combination between skim milk and CPO gives the highest protection results.

  8. Characterization of Whey Protein Oil-In-Water Emulsions with Different Oil Concentrations Stabilized by Ultra-High Pressure Homogenization

    Directory of Open Access Journals (Sweden)

    Essam Hebishy

    2017-02-01

    Full Text Available In this study, the effect of ultra-high-pressure homogenization (UHPH: 100 or 200 MPa at 25 °C, in comparison to colloid mill (CM: 5000 rpm at 20 °C and conventional homogenization (CH: 15 MPa at 60 °C, on the stability of oil-in-water emulsions with different oil concentrations (10, 30 or 50 g/100 g emulsified by whey protein isolate (4 g/100 g was investigated. Emulsions were characterized for their microstructure, rheological properties, surface protein concentration (SPC, stability to creaming and oxidative stability under light (2000 lux/m2. UHPH produced emulsions containing lipid droplets in the sub-micron range (100–200 nm and with low protein concentrations on droplet surfaces. Droplet size (d3.2, µm was increased in CH and UHPH emulsions by increasing the oil concentration. CM emulsions exhibited Newtonian flow behaviour at all oil concentrations studied; however, the rheological behaviour of CH and UHPH emulsions varied from Newtonian flow (n ≈ 1 to shear-thinning (n ˂ 1 and thixotropic behaviour in emulsions containing 50% oil. This was confirmed by the non-significant differences in the d4.3 (µm value between the top and bottom of emulsions in tubes left at room temperature for nine days and also by a low migration velocity measured with a Turbiscan LAB instrument. UHPH emulsions showed significantly lower oxidation rates during 10 days storage in comparison to CM and CH emulsions as confirmed by hydroperoxides and thiobarbituric acid-reactive substances (TBARS. UHPH emulsions treated at 100 MPa were less oxidized than those treated at 200 MPa. The results from this study suggest that UHPH treatment generates emulsions that have a higher stability to creaming and lipid oxidation compared to colloid mill and conventional treatments.

  9. Effect of prolonged exposure to sublethal concentrations of DDT and DDE on protein expression in human pancreatic beta cells

    Energy Technology Data Exchange (ETDEWEB)

    Pavlikova, Nela, E-mail: nela.pavlikova@lf3.cuni.cz [Department of Cell and Molecular Biology, Third Faculty of Medicine, Charles University, Prague (Czech Republic); Smetana, Pavel [Department of Cell and Molecular Biology, Third Faculty of Medicine, Charles University, Prague (Czech Republic); Halada, Petr [Laboratory of Molecular Structure Characterization, Institute of Microbiology, Academy of Sciences of the Czech Republic, Prague (Czech Republic); Kovar, Jan [Department of Cell and Molecular Biology, Third Faculty of Medicine, Charles University, Prague (Czech Republic)

    2015-10-15

    Pollution of the environment represents one of less explored potential reasons for the worldwide epidemic of type 2 diabetes. One of the most prevalent organochlorine pollutants remains the pesticide DDT and its degradation product DDE. Despite some epidemiologic correlations between levels of DDT and DDE in human organism and the prevalence of diabetes, there is almost no information about the exact targets of these compounds inside pancreatic beta cells. To detect functional areas of pancreatic beta cells that could be affected by exposure to DDT and DDE, we analyzed changes in protein expression in the NES2Y human pancreatic beta cell line exposed to three sublethal concentrations (0.1 μM, 1 μM, 10 μM) of DDT and DDE for 1 month. Protein separation and identification was achieved using high-resolution 2D-electrophoresis, computer analysis and mass spectrometry. With these techniques, four proteins were found downregulated after exposure to 10 μM DDT: three cytoskeletal proteins (cytokeratin 8, cytokeratin 18 and actin) and one protein involved in glycolysis (alpha-enolase). Two proteins were downregulated after exposure to 10 μM DDE: cytokeratin 18 and heterogenous nuclear ribonucleoprotein H1 (HNRH1). These changes correlate with previously described effects of other stress conditions (e.g. exposure to palmitate, hyperglycemia, imidazoline derivative, and cytokines) on protein expression in pancreatic beta cells. We conclude that cytoskeletal proteins and their processing, glucose metabolism, and mRNA processing may represent targets affected by exposure to conditions hostile to pancreatic beta cells, including exposure to DDT and DDE. - Highlights: • Epidemiologic studies connect pollution with incidence of diabetes mellitus. • We explored the effect of DDT and DDE on protein expression in the NES2Y pancreatic beta cell line. • One month exposure to three sublethal concentrations of DDT and DDE was employed. • Expression of alpha-enolase, actin

  10. Effect of prolonged exposure to sublethal concentrations of DDT and DDE on protein expression in human pancreatic beta cells

    International Nuclear Information System (INIS)

    Pavlikova, Nela; Smetana, Pavel; Halada, Petr; Kovar, Jan

    2015-01-01

    Pollution of the environment represents one of less explored potential reasons for the worldwide epidemic of type 2 diabetes. One of the most prevalent organochlorine pollutants remains the pesticide DDT and its degradation product DDE. Despite some epidemiologic correlations between levels of DDT and DDE in human organism and the prevalence of diabetes, there is almost no information about the exact targets of these compounds inside pancreatic beta cells. To detect functional areas of pancreatic beta cells that could be affected by exposure to DDT and DDE, we analyzed changes in protein expression in the NES2Y human pancreatic beta cell line exposed to three sublethal concentrations (0.1 μM, 1 μM, 10 μM) of DDT and DDE for 1 month. Protein separation and identification was achieved using high-resolution 2D-electrophoresis, computer analysis and mass spectrometry. With these techniques, four proteins were found downregulated after exposure to 10 μM DDT: three cytoskeletal proteins (cytokeratin 8, cytokeratin 18 and actin) and one protein involved in glycolysis (alpha-enolase). Two proteins were downregulated after exposure to 10 μM DDE: cytokeratin 18 and heterogenous nuclear ribonucleoprotein H1 (HNRH1). These changes correlate with previously described effects of other stress conditions (e.g. exposure to palmitate, hyperglycemia, imidazoline derivative, and cytokines) on protein expression in pancreatic beta cells. We conclude that cytoskeletal proteins and their processing, glucose metabolism, and mRNA processing may represent targets affected by exposure to conditions hostile to pancreatic beta cells, including exposure to DDT and DDE. - Highlights: • Epidemiologic studies connect pollution with incidence of diabetes mellitus. • We explored the effect of DDT and DDE on protein expression in the NES2Y pancreatic beta cell line. • One month exposure to three sublethal concentrations of DDT and DDE was employed. • Expression of alpha-enolase, actin

  11. The Sporothrix schenckii Gene Encoding for the Ribosomal Protein L6 Has Constitutive and Stable Expression and Works as an Endogenous Control in Gene Expression Analysis

    Directory of Open Access Journals (Sweden)

    Elías Trujillo-Esquivel

    2017-09-01

    Full Text Available Sporothrix schenckii is one of the causative agents of sporotrichosis, a worldwide-distributed mycosis that affects humans and other mammals. The interest in basic and clinical features of this organism has significantly increased in the last years, yet little progress in molecular aspects has been reported. Gene expression analysis is a set of powerful tools that helps to assess the cell response to changes in the extracellular environment, the genetic networks controlling metabolic pathways, and the adaptation to different growth conditions. Most of the quantitative methodologies used nowadays require data normalization, and this is achieved measuring the expression of endogenous control genes. Reference genes, whose expression is assumed to suffer minimal changes regardless the cell morphology, the stage of the cell cycle or the presence of harsh extracellular conditions are commonly used as controls in Northern blotting assays, microarrays, and semi-quantitative or quantitative RT-PCR. Since the biology of the organisms is usually species specific, it is difficult to find a reliable group of universal genes that can be used as controls for data normalization in experiments addressing the gene expression, regardless the taxonomic classification of the organism under study. Here, we compared the transcriptional stability of the genes encoding for elongation factor 1A, Tfc1, a protein involved in transcription initiation on Pol III promoters, ribosomal protein L6, histone H2A, β-actin, β-tubulin, glyceraldehyde 3-phosphate dehydrogenase, UAF30, the upstream activating factor 30, and the transcription initiation factor TFIID subunit 10, during the fungal growth in different culture media and cell morphologies. Our results indicated that only the gene encoding for the ribosomal protein L6 showed a stable and constant expression. Furthermore, it displayed not transcriptional changes when S. schenckii infected larvae of Galleria mellonella or

  12. Effect of dietary soybean oil and soybean protein concentration on the concentration of digestible amino acids in soybean products fed to growing pigs.

    Science.gov (United States)

    Cervantes-Pahm, S K; Stein, H H

    2008-08-01

    An experiment was conducted to measure the effect of adding soybean oil to soybean meal (SBM) and soy protein concentrate (SPC) on apparent (AID) and standardized (SID) ileal digestibility of CP and AA by growing pigs. A second objective was to compare AID and SID of AA in a new high-protein variety of full fat soybeans (FFSB) to values obtained in other soybean products. Commercial sources of FFSB (FFSB-CV), SBM, and SPC, and of a new high-protein variety of FFSB (FFSB-HP) were used in the experiment. Four diets were prepared using each soybean product as the sole source of CP and AA in 1 diet. Two additional diets were formulated by adding soybean oil (7.55 and 7.35%, respectively) to the diets containing SBM and SPC. A nitrogen-free diet was also used to measure basal endogenous losses of CP and AA. The 2 sources of FFSB were extruded at 150 degrees C before being used in the experiment. Seven growing barrows (initial BW = 26.2 kg) were prepared with a T-cannula in the distal ileum and allotted to a 7 x 7 Latin square design. Ileal digesta were collected from the pigs on d 6 and 7 of each period. All digesta samples were lyophilized and analyzed for DM, CP, AA, and chromium, and values for AID and SID of CP and AA were calculated. The addition of oil improved (P oil and SPC, but these values were lower (P oil. In conclusion, the addition of oil improved the SID of most AA in SBM and SPC fed to growing pigs, and the SID of AA in FFSB-HP were greater than in SBM and similar to the SID of AA in SBM with oil and in SPC.

  13. Protein concentrations of sweet soysauces from Rhizopus oryzae and R. oligosporus fermentation without moromi fermentation

    Directory of Open Access Journals (Sweden)

    NOOR SOESANTI HANDAJANI

    2007-07-01

    Full Text Available Soy sauce was produce from soybean that fermented with koji/tempeh fungi and thenfermented under salt solution or moromi fermentation. The objectives of this experiment was to compare of protein (total and soluble content of sweet soy sauce that produced from soybean fermented with Rhizopus oryzae and R. oligosporus without moromi fermentation to the sweet soysauce with moromi fermentation one. The total and soluble proteins of sweet soy sauces that produce from soybean without moromi fermentation were higher that sweet soy sauces that produce with moromi fermentation. Soluble protein of sweet soy sauce that produced from soybean fermented with R. oligosporus without moromi fermentation was 8.2% and meet to the highest quality of sweet soy sweet sauce based on Indonesia Industrial Standard. Soluble protein of sweet soy sauce that produced from soybean fermented with R. oryzae without moromi fermentation was 4.1% and meet to the medium quality of sweet soy sweet sauce based on Indonesia Industrial Standard.

  14. Selective separation and concentration of antihypertensive peptides from rapeseed protein hydrolysate by electrodialysis with ultrafiltration membranes.

    Science.gov (United States)

    He, Rong; Girgih, Abraham T; Rozoy, Elodie; Bazinet, Laurent; Ju, Xing-Rong; Aluko, Rotimi E

    2016-04-15

    Rapeseed protein isolate was subjected to alcalase digestion to obtain a protein hydrolysate that was separated into peptide fractions using electrodialysis with ultrafiltration membrane (EDUF) technology. The EDUF process (6h duration) led to isolation of three peptide fractions: anionic (recovered in KCl-1 compartment), cationic (recovered in KCl-2 compartment), and those that remained in the feed compartment, which was labeled final rapeseed protein hydrolysate (FRPH). As expected the KCl-1 peptides were enriched in negatively-charged (43.57%) while KCl-2 contained high contents of positively-charged (28.35%) amino acids. All the samples inhibited angiotensin converting enzyme (ACE) and renin activities in dose-dependent manner with original rapeseed protein hydrolysate having the least ACE-inhibitory IC50 value of 0.0932±0.0037 mg/mL while FRPH and KCl-2 had least renin-inhibitory IC50 values of 0.47±0.05 and 0.55±0.06 mg/mL, respectively. Six hours after oral administration (100 mg/kg body weight) to spontaneously hypertensive rats, the FRPH produced the maximum systolic blood pressure reduction of -51 mmHg. Copyright © 2015 Elsevier Ltd. All rights reserved.

  15. Registration of 'Bolles' hard red spring wheat with high grain protein concentration and superior baking quality

    Science.gov (United States)

    The hard red spring wheat market class in the U.S. commands the highest prices on the worldwide wheat markets because of its high protein content, strong gluten, and good baking properties. ‘Bolles’ (PI 678430), a hard red spring wheat cultivar, was released by the University of Minnesota Agricultu...

  16. Detection of protein concentrations using a pH-step titration method

    NARCIS (Netherlands)

    Kruise, J.; Kruise, J.; Eijkel, Jan C.T.; Bergveld, Piet

    1997-01-01

    A stimulus-response method based on the application of a pH step is proposed for the detection of protein immobilized in a membrane on top of an ion-sensitive field-effect transistor (ISFET). The ISFET response to a step-wise change in pH, applied at the interface between the membrane and the

  17. Cortical concentrations of metals and protein in the brain of two ...

    African Journals Online (AJOL)

    Metal pollution and exposure is an increasing global problem. One of the best ways to assess the impact of this problem on the brain is by using animals raised in such environments. As a prelude to this, the goat was used in this study to determine its normal metal and protein level in the brain. Ten metals and total crude ...

  18. Proteomic studies related to genetic determinants of variability in protein concentrations

    NARCIS (Netherlands)

    Horvatovich, Peter; Franke, Lude; Bischoff, Rainer

    2014-01-01

    Genetic variation has multiple effects on the proteome. It may influence the expression level of proteins, modify their sequences through single nucleotide polymorphisms, the occurrence of allelic variants, or alternative splicing (ASP) events. This perspective paper summarizes the major effects of

  19. Environmentally sensitive probes for monitoring protein-membrane interactions at nanomolar concentrations

    Czech Academy of Sciences Publication Activity Database

    Shvadchak, Volodymyr V.; Kucherak, Oleksandr; Afitska, Kseniia; Dziuba, D.; Yushchenko, Dmytro A.

    2017-01-01

    Roč. 1859, č. 5 (2017), s. 852-859 ISSN 0005-2736 Institutional support: RVO:61388963 Keywords : solvatochromic probes * fluorescence * protein-membrane interaction * affinity * binding stoichiometry * alpha-synuclein Subject RIV: CE - Biochemistry OBOR OECD: Biochemistry and molecular biology Impact factor: 3.498, year: 2016

  20. Unpredictably Stable

    DEFF Research Database (Denmark)

    Failla, Virgilio; Melillo, Francesca; Reichstein, Toke

    2014-01-01

    Is entrepreneurship a more stable career choice for high employment turnover individuals? We find that a transition to entrepreneurship induces a shift towards stayer behavior and identify job matching, job satisfaction and lock-in effects as main drivers. These findings have major implications...

  1. Effect of montmorillonite clay addition on the morphological and physical properties of Jatropha curcas L. and Glycine max L. protein concentrate films

    Science.gov (United States)

    Protein concentrates from jatropha (JPC) and soy seeds (SPC) were obtained by solubilization and acid precipitation of proteins. Films from JPC and SPC were prepared by the casting method, using two different montmorillonite (MMT) clay concentrations and plasticized with glycerol. Film properties we...

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

    NARCIS (Netherlands)

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

    2004-01-01

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

  3. Performance and meat quality traits of beef heifers fed with two levels of concentrate and ruminally undegradable protein.

    Science.gov (United States)

    Duarte, Marcio de Souza; Paulino, Pedro Veiga Rodrigues; Valadares Filho, Sebastião de Campos; Paulino, Mario Fonseca; Detmann, Edenio; Zervoudakis, Joanis Tilemahos; Monnerat, João Paulo Ismerio dos Santos; Viana, Gabriel da Silva; Silva, Luiz Henrique P; Serão, Nicola Vergara Lopes

    2011-04-01

    The effects of two levels of concentrate and ruminally undegradable protein (RUP) on performance, intake, digestibility, carcass characteristics, meat quality traits, and commercial cuts yield were assessed. Twenty crossbred heifers (240 kg average body weight) were used. At the beginning of the trial, four animals were slaughtered as reference group and the 16 remaining animals were randomly assigned to four treatments, in a 2 × 2 factorial design: two levels of concentrate (40% and 80%, dry matter (DM) basis) and two levels of RUP (48.79% and 27.19% of CP). At the end of the trial, all the animals were slaughtered. There was no interaction (P > 0.05) between concentrate and RUP levels. Dry matter intake and nutrients digestibility was not affected (P > 0.05) by RUP level. Heifers fed the highest RUP level had greater (P  0.05) DMI and ADG. Heifers fed diets with 80% concentrate had greater intake of TDN and EE, and lower intake of NDF (P RUP levels did not affect (P > 0.05) the carcass characteristics and carcass gain composition. Heifers fed 80% concentrate diets had larger (P  0.05) the composition of carcass gain. There was no effect (P > 0.05) of RUP and concentrate levels on meat quality traits and commercial cut yields.

  4. The effect of sub-minimum inhibitory concentration of ciprofloxacin concentrations on enteroaggregative Escherichia coli and the role of the surface protein dispersin

    Energy Technology Data Exchange (ETDEWEB)

    Mortensen, Ninell P [ORNL; Fowlkes, Jason Davidson [ORNL; Trevino-Dopatka, Sonia [ORNL; Maggart, Michael J [ORNL; Boisen, Nadia [University of Virginia School of Medicine; Doktycz, Mitchel John [ORNL; Nataro, James [University of Virginia School of Medicine; Allison, David P [ORNL

    2011-01-01

    Enteroaggregative Escherichia coli (EAEC) are bacterial pathogens that cause watery diarrhea, which is often persistent and can be inflammatory. The antibiotic ciprofloxacin is used to treat EAEC infections, but a full understanding of the antimicrobial effects of ciprofloxacin is needed for more efficient treatment of bacterial infections. In this study, it was found that sub-minimum inhibitory concentrations (sub-MICs) of ciprofloxacin had an inhibitory effect on EAEC adhesion to glass and mammalian HEp-2 cells. It was also observed that bacterial surface properties play an important role in bacterial sensitivity to ciprofloxacin. In an EAEC mutant strain where the hydrophobic positively charged surface protein dispersin was absent, sensitivity to ciprofloxacin was reduced compared with the wild-type strain. Identified here are several antimicrobial effects of ciprofloxacin at sub-MIC concentrations indicating that bacterial surface hydrophobicity affects the response to ciprofloxacin. Investigating the effects of sub-MIC doses of antibiotics on targeted bacteria could help to further our understanding of bacterial pathogenicity and elucidate future antibiotic treatment modalities.

  5. The effect of soy protein beverages on serum cell adhesion molecule concentrations in prehypertensive/stage 1 hypertensive individuals.

    Science.gov (United States)

    Dettmer, Michelle; Alekel, D Lee; Lasrado, Joanne A; Messina, Mark; Carriquiry, Alicia; Heiberger, Kevin; Stewart, Jeanne W; Franke, Warren

    2012-04-01

    Prehypertensive and hypertensive individuals are at increased risk of atherosclerotic cardiovascular disease (CVD), in part because hypertension contributes to endothelial dysfunction and increased cell adhesion molecule expression. Soy protein and isoflavones may favorably alter CVD risk factors, and hence the aim of this study was to determine whether intake of cow's milk compared with soy beverage prepared from whole soy bean (WSB) or soy protein isolate (SPI) would lower soluble cell adhesion molecule concentrations as a means of decreasing CVD risk. We enrolled healthy prehypertensive/stage 1 hypertensive men (n = 60; 18-63 years) and premenopausal women (n = 8; 20-48 years). Participants were randomized to 1 of 3 groups for 8 weeks: cow's milk (600 mL/d), SPI beverage (840 mL/d; 30.1 mg total isoflavones/d), or WSB beverage (840 mL/d; 91.4 mg total isoflavones/d). We measured soluble vascular cell adhesion molecule-1 (VCAM-1), intercellular cell adhesion molecule-1 (ICAM-1), and endothelial-leukocyte adhesion molecule-1 (E-selectin) concentrations at baseline and week 8. Soluble CAM concentrations were not altered by treatment and did not differ between prehypertensive and hypertensive participants. However, analysis of variance indicated a treatment × gender interaction (gender effect) for ICAM-1 (p = 0.0037) but not for E-selectin (p = 0.067) or VCAM-1 (p = 0.16). Men had higher concentrations of ICAM-1 and E-selectin, respectively, at baseline (p = 0.0071, p = 0.049) and week 8 (p = 0.0054, p = 0.038) than women did. Neither intake of cow's milk nor soy beverage for 8 weeks altered soluble CAM concentrations in prehypertensive/stage 1 hypertensive individuals, suggesting that neither type of beverage diminished atherosclerotic CVD risk in mildly hypertensive individuals by way of improving circulating CAM concentrations.

  6. Use of the Vettest 8008 and refractometry for determination of total protein, albumin, and globulin concentrations in feline effusions.

    Science.gov (United States)

    Papasouliotis, Kostas; Murphy, Kate; Dodkin, Steve; Torrance, Andy G

    2002-01-01

    Pleural and peritoneal effusion is a common clinical finding in feline practice. Determination of fluid albumin (ALB) and globulin (GLOB) concentrations in addition to total protein (TP) concentration can be helpful in diagnosing or ruling out certain diseases in cats, especially feline infectious peritonitis (FIP). The objective of this study was to compare effusion TP, ALB, and GLOB results obtained by a refractometer and a bench-top dry chemistry analyzer with those results obtained by a reference method. Twenty-six pleural and 14 peritoneal effusion samples were analyzed from 40 cats with various diseases. TP and ALB concentrations were determined by a reference automated wet chemistry analyzer (Kone Specific, Kone Instruments, Espoo, Finland), a bench-top dry chemistry analyzer (Vettest 8008, IDEXX Laboratories Ltd, Chalfont St Peter, UK), and a refractometer (Atago SPR-T2, Atago Co, Tokyo, Japan). GLOB, albumin to globulin (A/G) ratio, and globulins as a percentage of total proteins (GLOB%) were calculated. Results were analyzed by paired t tests, difference plots, and Deming s regression analysis. Correlation coefficients (r) for TP with Vettest versus Kone and refractometer versus Kone methods were.97 and.94, respectively. GLOB and GLOB% values were significantly higher and A/G ratios were significantly lower with Vettest versus Kone methods. Correlation coefficients for ALB, GLOB, GLOB% and A/G ratio with Vettest versus Kone methods were.86,.93,.82, and.73, respectively. Although correlation with other methods was good, the refractometer underestimated TP concentrations in 3 samples. The refractometer is an acceptable method for determination of TP concentration in feline effusions. The Vettest 8008 also is an acceptable method for the determination of TP and ALB concentrations, however, calculated A/G ratios obtained with the Vettest are unacceptable.

  7. 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.

    Science.gov (United States)

    Pihl, Tina H; Jacobsen, Stine; Olsen, Dorthe T; Højrup, Peter; Grosche, Astrid; Freeman, David E; Andersen, Pia H; Houen, Gunnar

    2017-06-01

    OBJECTIVE To purify and characterize equine vitamin D-binding protein (VDBP) from equine serum and to evaluate plasma concentrations of VDBP in healthy horses and horses with gastrointestinal injury or disease. ANIMALS 13 healthy laboratory animals (8 mice and 5 rabbits), 61 healthy horses, 12 horses with experimentally induced intestinal ischemia and reperfusion (IR), and 59 horses with acute gastrointestinal diseases. PROCEDURES VDBP was purified from serum of 2 healthy horses, and recombinant equine VDBP was obtained through a commercial service. Equine VDBP was characterized by mass 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 at predetermined time points in horses with IR and in horses with naturally occurring gastrointestinal diseases. RESULTS The working reference range for plasma VDBP concentration in healthy horses was 531 to 1,382 mg/L. Plasma VDBP concentrations were significantly decreased after 1 hour of ischemia in horses with IR, compared with values prior to induction of ischemia, and were significantly lower in horses with naturally occurring gastrointestinal diseases with a colic duration of < 12 hours than in healthy horses. CONCLUSIONS AND CLINICAL RELEVANCE Plasma VDBP concentrations were significantly decreased in horses with acute gastrointestinal injury or disease. Further studies and the development of a clinically relevant assay are needed to establish the reliability of VDBP as a diagnostic and prognostic marker in horses.

  8. Internal motion time scales of a small, highly stable and disulfide-rich protein: A 15N, 13C NMR and molecular dynamics study

    International Nuclear Information System (INIS)

    Guenneugues, Marc; Gilquin, Bernard; Wolff, Nicolas; Menez, Andre; Zinn-Justin, Sophie

    1999-01-01

    Motions of the backbone CαHα and threonine CβHβ bonds of toxin α were investigated using natural abundance 13C NMR and molecular dynamics. Measurement of the 13C longitudinal and transverse relaxation rates employed ACCORDION techniques together with coherence selection by pulsed field gradients and sensitivity enhancement through the use of preservation of equivalent pathway, thus allowing a considerable reduction of the required spectrometer time. 13C R1, R2, 1H → 13C NOE were obtained, as well as the variations of R1ρ(90 deg.) as a function of the rf field strength. These data were compared to those recorded by 1H and 15N NMR on a labelled sample of the toxin [Guenneugues et al. (1997) Biochemistry, 36, 16097-16108]. Both sets of data showed that picosecond to nanosecond time scale motions are well correlated to the secondary structure of the protein. This was further reinforced by the analysis of a 1 ns molecular dynamics simulation in water. Several CαHα and threonine CβHβ experimentally exhibit fast motions with a correlation time longer than 500 ps, that cannot be sampled along the simulation. In addition, the backbone exhibits motions on the microsecond to millisecond time scale on more than half of its length. Thus, toxin α, a highly stable protein (Tm=75 deg. C at acidic pH) containing 61 amino acids and 4 disulfides, shows important internal motions on time scales ranging from 0.1-0.5 ps, to 10-100 ps, 1 ns, and about 30 μs to 10 ms

  9. The effect of dietary protein and phosphorus on ammonia concentration and litter composition in broilers.

    Science.gov (United States)

    Ferguson, N S; Gates, R S; Taraba, J L; Cantor, A H; Pescatore, A J; Straw, M L; Ford, M J; Burnham, D J

    1998-08-01

    An experiment was conducted to determine whether broiler litter concentration of N and P and equilibrium NH3 gas concentration can be reduced by reducing dietary CP and P levels and supplementing with amino acids and phytase, respectively, without adversely affecting bird performance. Equilibrium NH3 gas concentration above the litter was measured. The experiment was divided into a starter period (1 to 21 d) and grower period (22 to 42 d), each having two different CP and P levels in a 2 x 2 factorial arrangement. The CP treatments consisted of a control with a mean CP of 204 and 202 g/kg for starter and grower periods, respectively, and a low CP diet with means of 188 and 183 g/kg, respectively, but with similar amino acid levels as the control. The P treatments comprised starter and grower control diets containing means of 6.7 and 6.3 g/kg P, respectively, and low P treatment means of 5.8 and 5.4 g/kg P supplemented with 1.0 g/kg phytase. Reducing starter diet CP by 16 g/kg reduced weight gain by 3.5% and, hence, body weight at 21 d of age, but did not affect feed intake or feed efficiency. Reducing P did not affect feed intake and weight gain, but improved feed efficiency by 2.0%. Responses in feed intake and efficiency to CP depended on the level of dietary P. For the grower period there were no significant differences in feed intake, weight gain, and feed efficiency, nor in body weight at 42 d of age, after correcting for 21-d body weight, between CP and P treatments. There were significant (P litter N and P concentrations, but not equilibrium NH3 gas concentration, moisture content, or pH, for low CP and P diets. Mean equilibrium NH3 gas concentration was 63 ppm. Litter N concentration was reduced 16.3% with the low CP diets, and litter P by 23.2% in low P treatments. The results suggest that dietary manipulation shows merit for reducing litter N and P concentrations while maintaining acceptable production performance from broilers.

  10. Wheat germ cell-free expression: Two detergents with a low critical micelle concentration allow for production of soluble HCV membrane proteins.

    Science.gov (United States)

    Fogeron, Marie-Laure; Badillo, Aurélie; Jirasko, Vlastimil; Gouttenoire, Jérôme; Paul, David; Lancien, Loick; Moradpour, Darius; Bartenschlager, Ralf; Meier, Beat H; Penin, François; Böckmann, Anja

    2015-01-01

    Membrane proteins are notoriously difficult to express in a soluble form. Here, we use wheat germ cell-free expression in the presence of various detergents to produce the non-structural membrane proteins 2, 4B and 5A of the hepatitis C virus (HCV). We show that lauryl maltose neopentyl glycol (MNG-3) and dodecyl octaethylene glycol ether (C12E8) detergents can yield essentially soluble membrane proteins at detergent concentrations that do not inhibit the cell-free reaction. This finding can be explained by the low critical micelle concentration (CMC) of these detergents, which keeps the monomer concentrations low while at the same time providing the necessary excess of detergent concentration above CMC required for full target protein solubilization. We estimate that a tenfold excess of detergent micelles with respect to the protein concentration is sufficient for solubilization, a number that we propose as a guideline for detergent screening assays. Copyright © 2014 Elsevier Inc. All rights reserved.

  11. Application of stable isotope tracer methods to studies of amino acid, protein, and energy metabolism in malnourished populations of developing countries. Report of an IAEA consultants' meeting held in Vienna, Austria, 14-16 December 1992

    International Nuclear Information System (INIS)

    1993-01-01

    A Consultants' Meeting convened by the IAEA in December 1992, made recommendations on the organization of a Co-ordinated Research Programme (CRP) using stable isotopic techniques for international comparative studies of amino acid, protein, and energy metabolism in chronically undernourished people. The CRP will use recent developments in stable isotope tracer techniques ( 13 C and 15 N) to assess the impact of infection in undernourished people on the kinetics of protein breakdown, protein synthesis, amino acid metabolism, and on the synthetic rates of selected plasma proteins. Studies will be conducted in developing countries, particularly in young children. The programme goals are to (i) elaborate methods and model protocols which can be implemented in developing countries to investigate the impact on protein metabolism of infection superimposed on chronic undernutrition; (ii) test they hypothesis that dietary requirements for protein and amino acids are related to the place of nutrition and are altered substantially when infection is superimposed on chronic undernutrition. When feasible, the primary focus on protein/amino acid metabolism will be extended to assessments of protein/energy interactions when H 2 18 O becomes more readily available and/or at research sites with indirect calorimetry equipment. The data generated should be appropriate as a basis for reevaluating amino acid/protein requirements in these populations. Refs

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

    DEFF Research Database (Denmark)

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

    2011-01-01

    pathway (LCP) recognition molecules and MAP-1. We expressed recombinant MAP-1 in CHO DG44 cells, developed a quantitative ELISA assay based on a MAP-1 specific monoclonal capture antibody and measured the serum levels in 100 Danish blood donors. In addition we assessed the association properties between......Recently, a novel protein named MBL/ficolin associated protein-1 (MAP-1) derived from the MASP1 gene through differential splicing was identified. In the present study, we established biochemical characteristics, determined the serum level and assessed the interactions between the lectin complement...... MAP-1 and Ficolin-2, -3 and MBL in serum using ELISA and density gradient ultra centrifugation. When recombinant MAP-1 was subjected to N-glycosidase F treatment the molecular mass decreased from ~45kDa to ~40kDa equivalent with the calculated molecular mass from the deduced amino acid sequence...

  13. Advanced control of dissolved oxygen concentration in fed batch cultures during recombinant protein production.

    Science.gov (United States)

    Kuprijanov, A; Gnoth, S; Simutis, R; Lübbert, A

    2009-02-01

    Design and experimental validation of advanced pO(2) controllers for fermentation processes operated in the fed-batch mode are described. In most situations, the presented controllers are able to keep the pO(2) in fermentations for recombinant protein productions exactly on the desired value. The controllers are based on the gain-scheduling approach to parameter-adaptive proportional-integral controllers. In order to cope with the most often appearing distortions, the basic gain-scheduling feedback controller was complemented with a feedforward control component. This feedforward/feedback controller significantly improved pO(2) control. By means of numerical simulations, the controller behavior was tested and its parameters were determined. Validation runs were performed with three Escherichia coli strains producing different recombinant proteins. It is finally shown that the new controller leads to significant improvements in the signal-to-noise ratio of other key process variables and, thus, to a higher process quality.

  14. Measurement of Endogenous versus Exogenous Formaldehyde-Induced DNA-Protein Crosslinks in Animal Tissues by Stable Isotope Labeling and Ultrasensitive Mass Spectrometry.

    Science.gov (United States)

    Lai, Yongquan; Yu, Rui; Hartwell, Hadley J; Moeller, Benjamin C; Bodnar, Wanda M; Swenberg, James A

    2016-05-01

    DNA-protein crosslinks (DPC) arise from a wide range of endogenous and exogenous chemicals, such as chemotherapeutic drugs and formaldehyde. Importantly, recent identification of aldehydes as endogenous genotoxins in Fanconi anemia has provided new insight into disease causation. Because of their bulky nature, DPCs pose severe threats to genome stability, but previous methods to measure formaldehyde-induced DPCs were incapable of discriminating between endogenous and exogenous sources of chemical. In this study, we developed methods that provide accurate and distinct measurements of both exogenous and endogenous DPCs in a structurally specific manner. We exposed experimental animals to stable isotope-labeled formaldehyde ([(13)CD2]-formaldehyde) by inhalation and performed ultrasensitive mass spectrometry to measure endogenous (unlabeled) and exogenous ((13)CD2-labeled) DPCs. We found that exogenous DPCs readily accumulated in nasal respiratory tissues but were absent in tissues distant to the site of contact. This observation, together with the finding that endogenous formaldehyde-induced DPCs were present in all tissues examined, suggests that endogenous DPCs may be responsible for increased risks of bone marrow toxicity and leukemia. Furthermore, the slow rate of DPC repair provided evidence for the persistence of DPCs. In conclusion, our method for measuring endogenous and exogenous DPCs presents a new perspective for the potential health risks inflicted by endogenous formaldehyde and may inform improved disease prevention and treatment strategies. Cancer Res; 76(9); 2652-61. ©2016 AACR. ©2016 American Association for Cancer Research.

  15. Performance and metabolite profile of dairy cows fed tropical grasses and concentrates containing crude protein with low or high degradability

    Directory of Open Access Journals (Sweden)

    Raphael dos Santos Gomes

    Full Text Available ABSTRACT Ten Holstein-Zebu crossbred cows distributed into two simultaneous Latin squares (5 × 5 as a 2 × 2 factorial arrangement formed by chopped sugarcane or elephant grass silage, both with high or low protein degradability supplements and a corn silage as a control treatment, were compared using orthogonal contrasts. The studied variables were the performance, plasma concentrations of urea-N, glucose, and creatinine, urine-N and milk urea-N, and the nychthemeral variation in NH3-N in the rumen fluid of dairy cows. Nutrient intake, milk production, and milk composition were affected by the treatments. The total mixed ration containing elephant grass silage combined with rumen undegradable protein (RUP provided balanced amounts of carbon and nitrogen in the rumen. This effect may explain the 18% increase in milk yield compared with the other treatments. The diurnal pattern of ruminal NH3-N was interpreted with a sinusoid model. In general, cows fed elephant grass silage exhibited higher concentrations of blood plasma and milk urea-N than animals fed sugarcane. The cows that consumed elephant grass silage with rumen degradable protein concentrate showed a higher milk urea-N compared with animals that consumed elephant grass silage with the RUP concentrate. The use of diets based on corn silage leads to a better use of nitrogen compounds because these diets resulted in lower levels of urea-N in the plasma, urine, and milk at the same level of milk production compared with diets containing elephant grass silage or chopped sugarcane as roughages. In sugarcane-based diets, even greater nitrogen losses in the urine are observed, despite the presence of readily fermentable carbohydrates in the diet.

  16. Airborne protein concentration: a key metric for type 1 allergy risk assessment-in home measurement challenges and considerations.

    Science.gov (United States)

    Tulum, Liz; Deag, Zoë; Brown, Matthew; Furniss, Annette; Meech, Lynn; Lalljie, Anja; Cochrane, Stella

    2018-01-01

    Exposure to airborne proteins can be associated with the development of immediate, IgE-mediated respiratory allergies, with genetic, epigenetic and environmental factors also playing a role in determining the likelihood that sensitisation will be induced. The main objective of this study was to determine whether airborne concentrations of selected common aeroallergens could be quantified in the air of homes using easily deployable, commercially available equipment and analytical methods, at low levels relevant to risk assessment of the potential to develop respiratory allergies. Additionally, air and dust sampling were compared and the influence of factors such as different filter types on allergen quantification explored. Low volume air sampling pumps and DUSTREAM ® dust samplers were used to sample 20 homes and allergen levels were quantified using a MARIA ® immunoassay. It proved possible to detect a range of common aeroallergens in the home with sufficient sensitivity to quantify airborne concentrations in ranges relevant to risk assessment (Limits of Detection of 0.005-0.03 ng/m 3 ). The methodology discriminates between homes related to pet ownership and there were clear advantages to sampling air over dust which are described in this paper. Furthermore, in an adsorption-extraction study, PTFE (polytetrafluoroethylene) filters gave higher and more consistent recovery values than glass fibre (grade A) filters for the range of aeroallergens studied. Very low airborne concentrations of allergenic proteins in home settings can be successfully quantified using commercially available pumps and immunoassays. Considering the greater relevance of air sampling to human exposure of the respiratory tract and its other advantages, wider use of standardised, sensitive techniques to measure low airborne protein concentrations and how they influence development of allergic sensitisation and symptoms could accelerate our understanding of human dose-response relationships

  17. Stable particles

    International Nuclear Information System (INIS)

    Samios, N.P.

    1993-01-01

    I have been asked to review the subject of stable particles, essentially the particles that eventually comprised the meson and baryon octets. with a few more additions -- with an emphasis on the contributions made by experiments utilizing the bubble chamber technique. In this activity, much work had been done by the photographic emulsion technique and cloud chambers-exposed to cosmic rays as well as accelerator based beams. In fact, many if not most of the stable particles were found by these latter two techniques, however, the forte of the bubble chamber (coupled with the newer and more powerful accelerators) was to verify, and reinforce with large statistics, the existence of these states, to find some of the more difficult ones, mainly neutrals and further to elucidate their properties, i.e., spin, parity, lifetimes, decay parameters, etc

  18. Impact of Ovine Whey Protein Concentrates and Clarification By-Products on the Yield and Quality of Whey Cheese

    Directory of Open Access Journals (Sweden)

    Carlos D. Pereira

    2007-01-01

    Full Text Available The effects of the addition of whey protein concentrates and clarification by-products obtained from ovine cheese whey and deproteinized whey (Sorelho on the yield and quality of the whey cheese (Requeijão have been evaluated. Whey protein concentrates were obtained by ultrafiltration of skimmed whey and Sorelho. The clarification by-products were obtained after the treatment of the skimmed whey and Sorelho by thermocalcic precipitation and microfiltration with two membranes (0.20 and 0.65 μm pore size. Next, the liophilization of the corresponding retentates was carried out. Each powder was added in three different mass ratios: 0.5, 1.0 and 1.5 %. The addition of the powders caused higher yields of the whey cheese – mainly the one with the additional whey powder – but it did not affect the strength of the products. The retention of water and other components of whey and milk in the whey cheese was influenced by the protein composition of the powders. In relation to colour parameters, the whey cheese manufactured with ultrafiltration and microfiltration retentate powders showed lower values of ligthness than the control whey cheese – mainly the whey cheese with 1.5 % of added powders. The microstructure constituted of small aggregates in the whey cheese manufactured with ultrafiltration and 0.20-μm microfiltration retentate powders and also by large, smooth structures in the other whey cheeses, especially in batches with added Sorelho powders.

  19. S100B Protein concentration in milk-formulas for preterm and term infants. Correlation with industrial preparation procedures.

    Science.gov (United States)

    Nigro, Francesco; Gagliardi, Luigi; Ciotti, Sabina; Galvano, Fabio; Pietri, Amedeo; Tina, Gabriella Lucia; Cavallaro, Daniela; La Fauci, Luca; Iacopino, Leonardo; Bognanno, Matteo; Li Volti, Giovanni; Scacco, Antonio; Michetti, Fabrizio; Gazzolo, Diego

    2008-05-01

    Human milk S100B protein possesses important neurotrophic properties. However, in some conditions human milk is substituted by milk formulas. The aims of the present study were: to assess S100B concentrations in milk formulas, to verify any differences in S100B levels between preterm and term infant formulas and to evaluate the impact of industrial preparation at predetermined phases on S100B content. Two different set of samples were tested: (i) commercial preterm (n = 36) and term (n = 36) infant milk formulas; ii) milk preterm (n = 10) and term infant (n = 10) formulas sampled at the following predetermined industrial preparation time points: skimmed cow milk (Time 0); after protein sources supplementation (Time 1); after pasteurization (Time 2); after spray-drying (Time 3). Our results showed that S100B concentration in preterm formulas were higher than in term ones (p 0.05) at Time 2, whereas a significant (p pasteurization but not spry-drying. New feeding strategies in preterm and term infants are therefore warranted in order to preserve S100B protein during industrial preparation.

  20. Determination of trace element concentrations and stable lead, uranium and thorium isotope ratios by quadrupole-ICP-MS in NORM and NORM-polluted sample leachates

    Energy Technology Data Exchange (ETDEWEB)

    Mas, J.L., E-mail: ppmasb@us.es [Dpto. Fisica Aplicada I, EPS, Universidad de Sevilla, 41012 Sevilla (Spain); Villa, M. [Servicio de Radioisotopos, Centro de Investigacion, Tecnologia e Innovacion (CITIUS), Universidad de Sevilla, Avda. Reina Mercedes 4b, 41012 Sevilla (Spain); Dpto. Fisica Aplicada II, ETS de Arquitectura, Universidad de Sevilla, Avda. Reina Mercedes 2, 41012 Sevilla (Spain); Hurtado, S. [Servicio de Radioisotopos, Centro de Investigacion, Tecnologia e Innovacion (CITIUS), Universidad de Sevilla, Avda. Reina Mercedes 4b, 41012 Sevilla (Spain); Garcia-Tenorio, R. [Dpto. Fisica Aplicada II, ETS de Arquitectura, Universidad de Sevilla, Avda. Reina Mercedes 2, 41012 Sevilla (Spain)

    2012-02-29

    Highlights: Black-Right-Pointing-Pointer Polluted sediment and NORM samples. Black-Right-Pointing-Pointer An efficient yet fast process allowing multi-parametric determinations in <3 days. Black-Right-Pointing-Pointer Trace element concentrations, Pb, Th and U isotope ratios with a single instrument. - Abstract: This work focuses on the monitoring of the potential pollution in scenarios that involve NORM-related industrial activities (environmental or in-door scenarios). The objective was to develop a method to determine extent and origin of the contamination, suitable for monitoring (i.e. simple, fast and economical) and avoiding the use of too many different instruments. It is presented a radiochemical method that allows the determination of trace element concentrations and {sup 206}Pb/{sup 207}Pb/{sup 208}Pb, {sup 238}U/{sup 234}U and {sup 232}Th/{sup 230}Th isotope ratios using a single sample aliquot and a single instrument (ICP-QMS). Eichrom UTEVA{sup Registered-Sign} extraction chromatography minicolumns were used to separate uranium and thorium in sample leachates. Independent ICP-MS determinations of uranium and thorium isotope ratios were carried out afterwards. Previously a small aliquot of the leachate was used for the determination of trace element concentrations and lead isotope ratios. Several radiochemical arrangements were tested to get maximum performances and simplicity of the method. The performances of the method were studied in terms of chemical yields of uranium and thorium and removal of the potentially interfering elements. The established method was applied to samples from a chemical industry and sediments collected in a NORM-polluted scenario. The results obtained from our method allowed us to infer not only the extent, but also the sources of the contamination in the area.

  1. The effect of bleaching agents on the degradation of vitamins and carotenoids in spray-dried whey protein concentrate.

    Science.gov (United States)

    Stout, M A; Park, C W; Drake, M A

    2017-10-01

    Previous research has shown that bleaching affects flavor and functionality of whey proteins. The role of different bleaching agents on vitamin and carotenoid degradation is unknown. The objective of this study was to determine the effects of bleaching whey with traditional annatto (norbixin) by hydrogen peroxide (HP), benzoyl peroxide (BP), or native lactoperoxidase (LP) on vitamin and carotenoid degradation in spray-dried whey protein concentrate 80% protein (WPC80). An alternative colorant was also evaluated. Cheddar whey colored with annatto (15 mL/454 L of milk) was manufactured, pasteurized, and fat separated and then assigned to bleaching treatments of 250 mg/kg HP, 50 mg/kg BP, or 20 mg/kg HP (LP system) at 50°C for 1 h. In addition to a control (whey with norbixin, whey from cheese milk with an alternative colorant (AltC) was evaluated. The control and AltC wheys were also heated to 50°C for 1 h. Wheys were concentrated to 80% protein by ultrafiltration and spray dried. The experiment was replicated in triplicate. Samples were taken after initial milk pasteurization, initial whey formation, after fat separation, after whey pasteurization, after bleaching, and after spray drying for vitamin and carotenoid analyses. Concentrations of retinol, a-tocopherol, water-soluble vitamins, norbixin, and other carotenoids were determined by HPLC, and volatile compounds were measured by gas chromatography-mass spectrometry. Sensory attributes of the rehydrated WPC80 were documented by a trained panel. After chemical or enzymatic bleaching, WPC80 displayed 7.0 to 33.3% reductions in retinol, β-carotene, ascorbic acid, thiamin, α-carotene, and α-tocopherol. The WPC80 bleached with BP contained significantly less of these compounds than the HP- or LP-bleached WPC80. Riboflavin, pantothenic acid, pyridoxine, nicotinic acid, and cobalamin concentrations in fluid whey were not affected by bleaching. Fat-soluble vitamins were reduced in all wheys by more than 90

  2. Concentration quenching of Eu2+ in a thermal-stable yellow phosphor Ca2BO3Cl:Eu2+ for LED application

    International Nuclear Information System (INIS)

    Zhang Xinguo; Zhang Jilin; Dong Zhiyue; Shi Jianxin; Gong Menglian

    2012-01-01

    A piece-shaped phosphor Ca 2 BO 3 Cl: Eu 2+ was synthesized by solid-state reaction method. This phosphor exhibited wide absorption in ultra-violet and visible range, and bright yellow emission band centering at 570 nm. The concentration quenching mechanism was verified to be a dipole–dipole interaction, and its critical transfer distance was about 17 Å by both calculated crystal structural method and experimental spectral method. This phosphor has a good thermal stability with a quenching temperature (T 1/2 ) of 200 °C. Yellow and white LEDs were fabricated with this phosphor and near UV chips, and the yellow LED has a high color purity of 97.0% and promising current tolerant property, while the white LED shows a luminous efficiency of 11.68 lm/W. - Highlights: ► Broadband excitable and strong yellow-emitting Ca 2 BO 3 Cl: Eu 2+ phosphor is obtained by solid state reaction. ► Concentration quenching mechanism of Ca 2 BO 3 Cl: Eu 2+ is dipole–dipole interaction. ► Quenching temperature (T 1/2 ) of Ca 2 BO 3 Cl: Eu 2+ is at 200 °C. ► As synthesized material can be used for LED phosphor application.

  3. Nutritional evaluation of phosphorylated pumpkin seed (Cucurbita moschata) protein concentrate in silver catfish Rhamdia quelen (Quoy and Gaimard, 1824).

    Science.gov (United States)

    Lovatto, Naglezi de Menezes; Goulart, Fernanda Rodrigues; de Freitas, Silvandro Tonetto; Mombach, Patricia Inês; Loureiro, Bruno Bianch; Bender, Ana Betine Beutinger; Boligon, Aline Augusti; Radünz Neto, João; da Silva, Leila Picolli

    2015-12-01

    An 8-week feeding trial was conducted to evaluate the effect of replacing fish meal with pumpkin seed meal (PSM) or phosphorylated protein concentrate of pumpkin seed meal (PPCPS) on growth and metabolic responses of silver catfish. Five isonitrogenous and isocaloric diets were formulated. Control diet contained fish meal as the main protein source. The treatment groups contained 25 and 50% of either PSM or PPCPS protein replaced the fishmeal protein. A total of 400 silver catfish, with initial mean weight of 24 ± 0.46 g, were distributed into 20 tanks. For data four orthogonal contrasts were applied: control diet versus PSM diets; control diets versus PPCPS diets; control versus other diets; PSM diets versus PPCPS diets. The results indicated that the fish fed PSM diets had lower weight gain when compared to either control diet or PPCPS. The PPCPS do not affect growth and protein efficiency ratio. Lower albumin contents were found for the control diet fish for the contrasts control diet versus PPCPS diet and control diet versus other diets. The hepatic ALAT enzyme activity was higher in the fish fed the control diet (P < 0.05). The hepatic ALP was most active in fish that received the PPCPS diets, when comparing control diet versus PPCPS diets and control diet versus other diets. The hepatosomatic index was higher for fish fed the PPCPS. Our results indicated that PPCPS presents relevant nutritional quality for fish and can replace the fish meal protein up to 50% without affecting growth, PER and intermediate metabolites in silver catfish.

  4. Ingestive behavior of grazing steers fed increasing levels of concentrate supplementation with different crude protein contents.

    Science.gov (United States)

    Mendes, Fabrício Bacelar Lima; Silva, Robério Rodrigues; de Carvalho, Gleidson Giordano Pinto; da Silva, Fabiano Ferreira; Lins, Túlio Otávio Jardim D Almeida; da Silva, Anderson Luiz Nascimento; Macedo, Venício; Abreu Filho, George; de Souza, Sinvaldo Oliveira; Guimarães, Joanderson Oliveira

    2015-02-01

    This study aimed to evaluate the ingestive behavior of steers on Brachiaria brizantha pasture fed diets with increasing levels of concentrate supplementation. Thirty-two crossbred steers in the finishing phase with average weight of 420 ± 8 kg were distributed in a completely randomized design with four treatments and eight replicates per treatment. Their behavior was assessed every 5 min for 24 h, in the middle of the experimental period. Variance and regression analyses at 0.05 % probability were adopted. The times spent grazing and ruminating reduced linearly (P <0.05), whereas the times spent at the trough (eating) and on other activities increased linearly (P <0.05) as the supplementation levels were elevated. The total feeding and chewing times decreased linearly (P <0.05) as the concentrate levels in the diet were elevated. By increasing the supplementation levels, the number of bites per day decreased linearly (P <0.05), and the feed efficiency of dry matter increased quadratically. Rumination efficiency of dry matter increased linearly (P <0.05) with increasing levels of concentrate supplementation. Grazing and rumination activities are reduced when the time devoted to other activities and at the trough are increased, as a result of the substitution effect.

  5. The effect of periodontal therapy on C-reactive protein, endothelial function, lipids and proinflammatory biomarkers in patients with stable coronary artery disease: study protocol for a randomized controlled trial.

    Science.gov (United States)

    Saffi, Marco Aurélio Lumertz; Furtado, Mariana Vargas; Montenegro, Márlon Munhoz; Ribeiro, Ingrid Webb Josephson; Kampits, Cassio; Rabelo-Silva, Eneida Rejane; Polanczyk, Carisi Anne; Rösing, Cassiano Kuchenbecker; Haas, Alex Nogueira

    2013-09-06

    Scarce information exists regarding the preventive effect of periodontal treatment in the recurrence of cardiovascular events. Prevention may be achieved by targeting risk factors for recurrent coronary artery disease (CAD) in patients with previous history of cardiovascular events. The aim of this trial is to compare the effect of two periodontal treatment approaches on levels of C-reactive protein, lipids, flow-mediated dilation and serum concentrations of proinflammatory and endothelial markers in stable CAD patients with periodontitis over a period of 12 months. This is a randomized, parallel design, examiner blinded, controlled clinical trial. Individuals from both genders, 35 years of age and older, with concomitant diagnosis of CAD and periodontitis will be included. CAD will be defined as the occurrence of at least one of the following events 6 months prior to entering the trial: documented history of myocardial infarction; surgical or percutaneous myocardial revascularization and lesion >50% in at least one coronary artery assessed by angiography; presence of angina and positive noninvasive testing of ischemia. Diagnosis of periodontitis will be defined using the CDC-AAP case definition (≥2 interproximal sites with clinical attachment loss ≥6 mm and ≥1 interproximal site with probing depth ≥5 mm). Individuals will have to present at least ten teeth present to be included. One hundred individuals will be allocated to test (intensive periodontal treatment comprised by scaling and root planing) or control (community periodontal treatment consisting of one session of supragingival plaque removal only) treatment groups. Full-mouth six sites per tooth periodontal examinations and subgingival biofilm samples will be conducted at baseline, 3, 6 and 12 months after treatment. The primary outcome of this study will be C-reactive protein changes over time. Secondary outcomes include levels of total cholesterol, LDL-C, HDL-C, triglycerides, IL-1β, IL-6, TNF

  6. Comparison of refractometry and biuret assay for measurement of total protein concentration in canine abdominal and pleural fluid specimens.

    Science.gov (United States)

    Rose, Alexandra; Funk, Deborah; Neiger, Reto

    2016-04-01

    To compare total protein (TP) concentrations in canine pleural and abdominal fluid specimens as measured by refractometry and biuret assay. Diagnostic test evaluation. Data regarding 92 pleural and 148 abdominal fluid specimens from dogs with various diseases. TP concentrations in fluid specimens as measured by refractometry and biuret assay were recorded. Strength of association between sets of measurements was assessed by Spearman rank correlations and Bland-Altman plots. Optimal concentration cutoff for diagnostic discrimination between exudate and nonexudate was identified by construction of receiver operating characteristic curves. Median TP concentration in pleural fluid specimens was 2.7 g/dL (range, 0.3 to 4.8 g/dL) for refractometry and 2.9 g/dL (range, 0.7 to 5.8 g/dL) for biuret assay. Median TP concentration in abdominal fluid specimens was 3.5 g/dL (range, 0.1 to 6.0 g/dL) for refractometry and 3.5 g/dL (range, 0.6 to 5.7 g/dL) for biuret assay. Correlation was significant between refractometric and biuret results for pleural (ρ = 0.921) and abdominal (ρ = 0.908) fluid. Bland-Altman plots revealed bias of -0.18 g/dL for pleural fluid and -0.03 g/dL for abdominal fluid for refractometry versus biuret assay. With a TP concentration of ≥ 3 g/dL used to distinguish exudate from nonexudate, sensitivity of refractometry was 77% for pleural fluid and 80% for abdominal fluid. Specificity was 100% and 94%, respectively. Refractometry yielded acceptable results for measurement of TP concentration in canine pleural and abdominal fluid specimens, providing a more rapid and convenient method than biuret assay.

  7. Oral contraceptives increase insulin-like growth factor binding protein-1 concentration in women with polycystic ovarian disease.

    Science.gov (United States)

    Suikkari, A M; Tiitinen, A; Stenman, U H; Seppälä, M; Laatikainen, T

    1991-05-01

    Insulin-like growth factor-I (IGF-I) stimulates ovarian androgen production. Insulin-like growth factor binding protein-1 (IGFBP-1) inhibits IGF actions in vitro. To investigate the effect of oral contraceptive (OC) pills, given for 3 months, on serum gonadotropin, androgen, IGF-I, and IGFBP-1 concentrations, and glucose tolerance in seven women with polycystic ovarian disease (PCOD) and in five healthy control subjects. Seven women with PCOD and five healthy control subjects. An oral glucose tolerance test (OGTT) was performed before and after treatment with OC. After treatment with OC, serum luteinizing hormone, androstenedione, and free testosterone levels decreased, and sex hormone-binding globulin concentration increased in the women with PCOD as well as in the control subjects. The cumulative response of serum insulin to OGTT was larger in the women with PCOD than in the control subjects both before and after treatment. Serum IGF-I concentration, which was unchanged during OGTT, decreased from basal level of 326 +/- 70 micrograms/L to 199 +/- 28 micrograms/L after treatment with OC in the women with PCOD, whereas no change was found in the control subjects (from 235 +/- 11 micrograms/L to 226 +/- 11 micrograms/L). Treatment with OC caused an increase of the mean basal IGFBP-1 concentration from 24 +/- 7 micrograms/L to 73 +/- 14 micrograms/L in the women with PCOD. This increase was constant during the OGTT. In the control subjects, treatment with OC did not result in any significant change in IGFBP-1 concentrations (from 44 +/- 11 micrograms/L to 61 +/- 9 micrograms/L). The combination of decreased total IGF-I concentration and increased IGFBP-1 concentration induced by OC may decrease ovarian androgen production in PCOD.

  8. Effects of different protein concentrations on longevity and feeding behavior of two adult populations of Ceratitis capitata Wiedemann (Diptera: Tephritidae)

    International Nuclear Information System (INIS)

    Placido-Silva, Maria do Carmo; Silva Neto, Alberto M. da; Joachim-Bravo, Iara S.; Zucoloto, Fernando S.

    2006-01-01

    The effects of protein intake on two adult male and female populations of Ceratitis capitata Wiedemann were assessed. One population consisted of flies reared for twenty years in the laboratory (Lab-pop); the other population consisted both of flies reared in the laboratory for approximately fifteen years and of the periodically introduced wild flies (Hybrid-pop). Three diets were tested: a no-yeast diet and two diets containing yeast (protein source) at the concentrations 6.5 g or 1.5 g per 100 ml diet. The parameters analyzed were: adult longevity, diet intake with and without yeast, and discrimination threshold for yeast. Protein intake increased Lab-pop adult longevity and did not affect longevity of the Hybrid-pop. Longevity in each population was similar for males and females fed on the same diet. Food behavior were similar for male and female adults of both populations; all preferred diets containing protein (yeast). Males and females in both populations ingested similar amounts of each diet. The discrimination threshold for yeast was similar for all males (0.5 g yeast/100 ml diet); Lab-pop females were able to detect the presence of smaller quantities of yeast in their diet, thus having a higher discrimination capacity (0.4 g/100 ml diet) as compared to the Hybrid-pop females (0.6 g/ 100 ml diet). (author)

  9. Glycolysis and gluconeogenesis in the liver of catfish fed with different concentrations of proteins, lipids and carbohydrates

    Directory of Open Access Journals (Sweden)

    J.F.B. Melo

    Full Text Available ABSTRACT The activities of enzymes from a number of metabolic pathways have been used as a tool to evaluate the best use of nutrients on fish performance. In the present study the catfish Rhamdia quelen was fed with diets containing crude protein-lipid-carbohydrate (% as follows: treatment (T T1: 19-19-44; T2: 26-15-39; T3: 33-12-33; and T4: 40-10-24. The fish were held in tanks of re-circulated, filtered water with controlled temperature and aeration in 2000L experimental units. The feeding experiment lasted 30 days. The following enzymes of the carbohydrate metabolism were determined: Glucokinase (GK, Phosphofructokinase 1 (PFK-1, Pyruvate kinase (PK, Fructose-1,6-biphosphatase 1 (FBP-1. The activities of 6 phosphogluconate dehydrogenase (6PGDH and glucose 6 phosphate dehydrogenase (G6PDH were also assayed. The influence of nutrient levels on the enzyme activities is reported. The increase of dietary protein plus reduction of carbohydrates and lipids attenuates the glycolytic activity and induces hepatic gluconeogenesis as a strategy to provide metabolic energy from amino acids. The fish performance was affected by the concentrations of protein, lipid and carbohydrates in the diet. The greatest weight gain was obtained in fish fed diet T4 containing 40.14% of crude protein, 9.70% of lipids, and 24.37% of carbohydrate, respectively.

  10. Digestibility of Quinoa (Chenopodium quinoa Willd.) Protein Concentrate and Its Potential to Inhibit Lipid Peroxidation in the Zebrafish Larvae Model.

    Science.gov (United States)

    Vilcacundo, R; Barrio, D; Carpio, C; García-Ruiz, A; Rúales, J; Hernández-Ledesma, B; Carrillo, W

    2017-09-01

    Quinoa protein concentrate (QPC) was extracted and digested under in vitro gastrointestinal conditions. The protein content of QPC was in the range between 52.40 and 65.01% depending on the assay used. Quinoa proteins were almost completely hydrolyzed by pepsin at pH of 1.2, 2.0, and 3.2. At high pH, only partial hydrolysis was observed. During the duodenal phase, no intact proteins were visible, indicating their susceptibility to the in vitro simulated digestive conditions. Zebrafish larvae model was used to evaluate the in vivo ability of gastrointestinal digests to inhibit lipid peroxidation. Gastric digestion at pH 1.2 showed the highest lipid peroxidation inhibition percentage (75.15%). The lipid peroxidation activity increased after the duodenal phase. The digest obtained at the end of the digestive process showed an inhibition percentage of 82.10%, comparable to that showed when using BHT as positive control (87.13%).

  11. Putative midkine family protein up-regulation in Patella caerulea (Mollusca, Gastropoda) exposed to sublethal concentrations of cadmium

    International Nuclear Information System (INIS)

    Vanucci, Silvana; Minerdi, Daniela; Kadomatsu, Kenji; Mengoni, Alessio; Bazzicalupo, Marco

    2005-01-01

    A cDNA sequence of a putative midkine (MK) family protein was identified and characterised in the mollusc Patella caerulea. The midkine family consists of two members, midkine and pleiotrophin (PTN), and it is one of the recently discovered cytokines. Our results show that this putative midkine protein is up-regulated in specimens of P. caerulea exposed to sublethal cadmium concentrations (i.e. 0.5 and 1 mg l -1 Cd) over a 10-day exposure period. Semiquantitative RT-PCR and quantitative Real time RT-PCR estimations indicate elevated expression of midkine mRNA in exposed specimens compared to controls. Moreover, RT-PCR Real time values were higher in the viscera (here defined as the part of the soft tissue including digestive gland plus gills) than in the foot (i.e. foot plus head plus heart) of the limpets. At present, information on the functional signalling significance of the midkine family proteins suggests that the up-regulation of P. caerulea putative midkine family protein is a distress signal likely with informative value on health status of the organism and with potential prognostic capability

  12. Association between excess body weight and urine protein concentration in healthy dogs.

    Science.gov (United States)

    Tefft, Karen M; Shaw, Darcy H; Ihle, Sherri L; Burton, Shelley A; Pack, LeeAnn

    2014-06-01

    Markedly overweight people can develop progressive proteinuria and kidney failure secondary to obesity-related glomerulopathy (ORG). Glomerular lesions in dogs with experimentally induced obesity are similar to those in people with ORG. The aim of this study was to evaluate if urine protein and albumin excretion is greater in overweight and obese dogs than in dogs of ideal body condition. Client-owned dogs were screened for underlying health conditions. These dogs were assigned a body condition score (BCS) using a 9-point scoring system. Dogs with a BCS of ≥ 6 were classified as being overweight/obese, and dogs with a BCS of 4 or 5 were classified as being of ideal body weight. The urine protein:creatinine ratio (UPC) and urine albumin:creatinine ratio (UAC) were then determined, and compared between 20 overweight/obese dogs and 22 ideal body weight control dogs. Median UPC (0.04 [range, 0.01-0.14; interquartile range, 0.07]) and UAC (0.41 [0-10.39; 3.21]) of overweight/obese dogs were not significantly different from median UPC (0.04 [0.01-0.32; 0.07]) and UAC (0.18 [0-7.04; 1.75]) in ideal body weight dogs. Clinicopathologic abnormalities consistent with ORG were absent from overweight/obese dogs in this study. © 2014 American Society for Veterinary Clinical Pathology and European Society for Veterinary Clinical Pathology.

  13. Gamma and x radiation and thermal neutrons effects in lens solutions and the relation with proteins concentration

    International Nuclear Information System (INIS)

    Ramirez A, M.; Alarcon C, A.

    1996-01-01

    Radiation effects have been studied irradiating porcine lens solutions with doses which range between 52 Gy to 1042 Gy in the case of x-rays (30 kVp), 631 Gy to 4001 Gy in the case of 60 Co gamma rays and 314 Gy to 7596 Gy for thermal neutrons. The optics density time variation of solutions was determined using a Spectronic-501 spectrophotometer, and with this data an equation which describes the behavior in the mentioned cases was found. A phenomenological model is postulated which connects the optical time variation density increment macroscopic effect with proteins concentration in the crystalline lens obtaining relative biological effectiveness using the supra-molecular aggregate formation due to the denaturalization and destruction of lens proteins by radiation criteria. (authors). 5 refs., 3 figs

  14. [Influence of extremely low frequency magnetic field on total protein and -sh groups concentrations in liver homogenates].

    Science.gov (United States)

    Ciejka, Elżbieta; Kowalczyk, Agata; Gorąca, Anna

    2014-01-01

    Free radicals are atoms, molecules or their fragments, whose excess leads to the development of oxidative stress, the cause of many neoplastic, neurodegenerative and inflammatory diseases, as well as aging of organisms. Industrial pollution, tobacco smoke, ionizing radiation, ultrasound and magnetic fields are the major exogenous sources of free radicals. The low frequency mag- netic field is commonly applied in physiotherapy. The aim of the present study was to evaluate the effect of extremely low frequency magnetic field (1L.F-MF) on the concentration ofsullhydryl groups (-SH) and proteins in liver tissues of experimental animals de- pending on the time of exposure to the field. Twenty one Sprague-D)awley male rats, aged 3-4 months were randomly divided into 3 experimental groups (each containing 7 animals): controls (group I), the rats exposed to IEI.F-MF of 40 Hz, 7 mT (this kind of the ELF-MF is mostly used in magnetotherapy), 30 min/day for 2 weeks (group II) and the rats exposed to 40 Hz, 7 mT for 60 min/day for 2 weeks (group III). The concentrations of proteins and sulfhydryl groups in the liver tissues were determined after