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Sample records for stable protein concentrates

  1. Stable Binding of Alternative Protein-enriched Food Matrices with Concentrated Cranberry Bioflavonoids for Functional Food Applications

    Science.gov (United States)

    Grace, Mary H.; Guzman, Ivette; Roopchand, Diana E.; Moskal, Kristin; Cheng, Diana M.; Pogrebnyak, Natasha; Raskin, Ilya; Howell, Amy; Lila, Mary Ann

    2013-01-01

    Defatted soy flour (DSF), soy protein isolate (SPI), hemp protein isolate (HPI), medium roast peanut flour (MPF) and pea protein isolate (PPI) stably bind and concentrate cranberry (CB) polyphenols, creating protein/polyphenol-enriched matrices. Proanthocyanidins (PAC) in the enriched matrices ranged from 20.75 mg/g (CB-HPI) to 10.68 mg/g (CB-SPI). Anthocyanins (ANC) ranged from 3.19 mg/g (CB-DSF) to 1.68 mg/g (CB-SPI), while total phenolics (TP) ranged from 37.61 mg/g (CB-HPI) to 21.29 mg/g (CB-SPI). LC-MS indicated that the enriched matrices contained all identifiable ANC, PAC and flavonols present in CB juice. Complexation with SPI stabilized and preserved the integrity of the CB polyphenolic components for at least 15 weeks at 37 °C. PAC isolated from enriched matrices demonstrated comparable anti-adhesion bioactivity to PAC isolated directly from CB juice (MIC 0.4 to 0.16 mg/mL), indicating their potential utility for maintenance of urinary tract health. Approximately 1.0 g of polyphenol-enriched matrix delivered the same amount of PAC available in one cup (300 mL) of commercial CB juice cocktail; which has been shown clinically to be the prophylactic dose for reducing recurring urinary tract infections. CB-SPI inhibited gram- positive and gram-negative bacterial growth. Nutritional and sensory analyses indicated that the targeted CB-matrix combinations have high potential for incorporation in functional food formulations. PMID:23786629

  2. Concentration of stable elements in food products

    Energy Technology Data Exchange (ETDEWEB)

    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.

  3. Chemically Stable Lipids for Membrane Protein Crystallization

    Energy Technology Data Exchange (ETDEWEB)

    Ishchenko, Andrii; Peng, Lingling; Zinovev, Egor; Vlasov, Alexey; Lee, Sung Chang; Kuklin, Alexander; Mishin, Alexey; Borshchevskiy, Valentin; Zhang, Qinghai; Cherezov, Vadim (MIPT); (USC); (Scripps)

    2017-05-01

    The lipidic cubic phase (LCP) has been widely recognized as a promising membrane-mimicking matrix for biophysical studies of membrane proteins and their crystallization in a lipidic environment. Application of this material to a wide variety of membrane proteins, however, is hindered due to a limited number of available host lipids, mostly monoacylglycerols (MAGs). Here, we designed, synthesized, and characterized a series of chemically stable lipids resistant to hydrolysis, with properties complementary to the widely used MAGs. In order to assess their potential to serve as host lipids for crystallization, we characterized the phase properties and lattice parameters of mesophases made of two most promising lipids at a variety of different conditions by polarized light microscopy and small-angle X-ray scattering. Both lipids showed remarkable chemical stability and an extended LCP region in the phase diagram covering a wide range of temperatures down to 4 °C. One of these lipids has been used for crystallization and structure determination of a prototypical membrane protein bacteriorhodopsin at 4 and 20 °C.

  4. Radioactive and stable elements' concentration in medicinal plants from Brazil

    International Nuclear Information System (INIS)

    Sussa, F.V.; Silva, P.S.C.; Damatto, S.R.; Favaro, D.I.T.; Mazzilli, B.P.

    2009-01-01

    Since the early days of mankind, plants have been used as food and for medicinal purposes. Still, little information exists in literature about the activity concentration of 238 U and 232 Th decay products, as well as stable element concentrations in Brazilian plants. Activity concentrations of 226 Ra, 228 Ra and 210 Pb, and chemical concentrations of As, Ba, Br, Cs, Co, Cr, Cu, Eu, Fe, Hf, La, Lu, Rb, Sb, Sc, Sm, Ta, Tb, Yb, Zn and Zr were determined in ten samples commonly used in Brazilian medicinal plants. (author)

  5. 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

  6. Protein body formation in stable transgenic tobacco expressing elastin-like polypeptide and hydrophobin fusion proteins.

    Science.gov (United States)

    Gutiérrez, Sonia P; Saberianfar, Reza; Kohalmi, Susanne E; Menassa, Rima

    2013-05-10

    Plants are recognized as an efficient and inexpensive system to produce valuable recombinant proteins. Two different strategies have been commonly used for the expression of recombinant proteins in plants: transient expression mediated by Agrobacterium; or stable transformation of the plant genome. However, the use of plants as bioreactors still faces two main limitations: low accumulation levels of some recombinant proteins and lack of efficient purification methods. Elastin-like polypeptide (ELP), hydrophobin I (HFBI) and Zera® are three fusion partners found to increase the accumulation levels of recombinant proteins and induce the formation of protein bodies (PBs) in leaves when targeted to the endoplasmic reticulum (ER) in transient expression assays. In this study the effects of ELP and HFBI fusion tags on recombinant protein accumulation levels and PB formation was examined in stable transgenic Nicotiana tabacum. The accumulation of recombinant protein and PB formation was evaluated in two cultivars of Nicotiana tabacum transformed with green fluorescent protein (GFP) fused to ELP or HFBI, both targeted and retrieved to the ER. The ELP and HFBI tags increased the accumulation of the recombinant protein and induced the formation of PBs in leaves of stable transgenic plants from both cultivars. Furthermore, these tags induced the formation of PBs in a concentration-dependent manner, where a specific level of recombinant protein accumulation was required for PBs to appear. Moreover, agro-infiltration of plants accumulating low levels of recombinant protein with p19, a suppressor of post-transcriptional gene silencing (PTGS), increased accumulation levels in four independent transgenic lines, suggesting that PTGS might have caused the low accumulation levels in these plants. The use of ELP and HFBI tags as fusion partners in stable transgenic plants of tobacco is feasible and promising. In a constitutive environment, these tags increase the accumulation levels

  7. 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.

  8. 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.

  9. Characterization of a novel stable biocatalyst obtained by protein engineering

    NARCIS (Netherlands)

    Van den Burg, B; de Kreij, A; Van der Veek, P; Mansfeld, J; Venema, G

    Protein engineering is a powerful tool for the improvement of the properties of biocatalysts, Previously we have applied protein engineering technologies to obtain an extremely stable variant of the thermolysin-like protease from Bacillus stearothermophilus [Van den Burg, Vriend, Veltman, Venema and

  10. Protein solutions: concentration by a rapid method.

    Science.gov (United States)

    Blatt, W F; Feinberg, M P; Hopfenberg, H B; Saravis, C A

    1965-10-08

    Protein solutions were concentrated, with no evidence of denaturation, by means of membranes formed from the complex interaction product of polyanions and polycations (Diaplex). The technique is considerably more rapid than conventional ultrafiltration with Visking tubing.

  11. Stable isotope, site-specific mass tagging for protein identification

    Science.gov (United States)

    Chen, Xian

    2006-10-24

    Proteolytic peptide mass mapping as measured by mass spectrometry provides an important method for the identification of proteins, which are usually identified by matching the measured and calculated m/z values of the proteolytic peptides. A unique identification is, however, heavily dependent upon the mass accuracy and sequence coverage of the fragment ions generated by peptide ionization. The present invention describes a method for increasing the specificity, accuracy and efficiency of the assignments of particular proteolytic peptides and consequent protein identification, by the incorporation of selected amino acid residue(s) enriched with stable isotope(s) into the protein sequence without the need for ultrahigh instrumental accuracy. Selected amino acid(s) are labeled with .sup.13C/.sup.15N/.sup.2H and incorporated into proteins in a sequence-specific manner during cell culturing. Each of these labeled amino acids carries a defined mass change encoded in its monoisotopic distribution pattern. Through their characteristic patterns, the peptides with mass tag(s) can then be readily distinguished from other peptides in mass spectra. The present method of identifying unique proteins can also be extended to protein complexes and will significantly increase data search specificity, efficiency and accuracy for protein identifications.

  12. 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.

  13. 21 CFR 184.1979c - Whey protein concentrate.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Whey protein concentrate. 184.1979c Section 184... Listing of Specific Substances Affirmed as GRAS § 184.1979c Whey protein concentrate. (a) Whey protein... the finished dry product contains not less than 25 percent protein. Whey protein concentrate is...

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

    Energy Technology Data Exchange (ETDEWEB)

    Kubala-Kukus, A. [Institute of Physics, Swietokrzyska Academy, Swietokrzyska 15, 25-406 Kielce (Poland)]. E-mail: Aldona.Kubala-Kukus@pu.kielce.pl; Kuternoga, E. [Institute of Physics, Swietokrzyska Academy, Swietokrzyska 15, 25-406 Kielce (Poland); Braziewicz, J. [Institute of Physics, Swietokrzyska Academy, Swietokrzyska 15, 25-406 Kielce (Poland); Pajek, M. [Institute of Physics, Swietokrzyska Academy, Swietokrzyska 15, 25-406 Kielce (Poland)

    2004-10-08

    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{sup 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.

  15. 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

  16. 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)

  17. Vaspin plasma concentrations and mRNA expressions in patients with stable and unstable angina pectoris.

    Science.gov (United States)

    Li, Hai Ling; Peng, Wen Hui; Cui, Shi Tao; Lei, Hou; Wei, Yi Dong; Li, Wei Ming; Xu, Ya Wei

    2011-09-01

    Vaspin was a recently identified adipokine, playing a protective role in many metabolic diseases. The present study aimed to investigate the association between vaspin plasma level and stable angina pectoris (SAP) and unstable angina pectoris (UAP). A total of 88 patients with angiographically-proved coronary artery disease (CAD) (SAP 47, UAP 41) and 103 control subjects without cardiovascular diseases were enrolled in this study. Circulating vaspin, mRNA expression of vaspin in peripheral blood mononuclear cells (PBMC), clinical parameters, lipid profile and high-sensitivity C-reactive protein (hsCRP) were assayed. The severity of CAD was also assessed according to the number of vessels diseased. There are significant differences in circulating vaspin levels and mRNA levels of PBMC between SAP and UAP groups (SAP 0.91±0.95 ng/mL and UAP 0.43±0.38 ng/mL, p<0.01 in circulating vaspin level; SAP 1.19±0.85 and UAP 0.82±0.56, p<0.05 in mRNA level of PBMC). An inverse correlation between the number of diseased vessels and plasma vaspin concentration was observed (r=-0.350, p<0.01) in the CAD group. Construction of receiver operating characteristic curves confirmed that vaspin plasma concentrations significantly differentiated CAD patients (area under the curve=0.684, p<0.001), as well as UAP (area under the curve=0.640, p<0.05). Decreased vaspin plasma levels and mRNA levels in PBMC were observed in patients with UAP. Low vaspin concentrations correlate with CAD severity. The findings suggested that vaspin could serve as a novel biomarker of CAD as well as UAP.

  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. Enhanced TOF-SIMS imaging of a micropatterned protein by stable isotope protein labeling.

    Science.gov (United States)

    Belu, A M; Yang, Z; Aslami, R; Chilkoti, A

    2001-01-15

    Patterning of biomolecules on surfaces is an increasingly important technological goal. Because the fabrication of biomolecule arrays often involves stepwise, spatially resolved derivatization of surfaces, spectroscopic imaging of these arrays is important in their fabrication and optimization. Although imaging time-of-flight secondary ion mass spectrometry (TOF-SIMS) is a powerful method for spatially resolved surface analysis, TOF-SIMS images of micropatterned proteins on organic substrates can be difficult to acquire, because of the lack of high intensity, protein-specific molecular ions that are essential for imaging under static conditions. In contrast, low-mass ions are of suitable intensity for imaging, but can originate from different chemical species on the surface. A potential solution to this problem is to utilize stable isotope labeled proteins, an approach that has heretofore not been explored in TOF-SIMS imaging of micropatterned proteins and peptides. To investigate the feasibility of stable isotope enhanced TOF-SIMS imaging of proteins, we synthesized 15N-labeled streptavidin by labeling of the protein during expression from a recombinant gene. The spatial distribution of streptavidin bound to biotin micropatterns, fabricated on a polymer and on a self-assembled monolayer on gold, was imaged by TOF-SIMS. Imaging of high-intensity, low-m/z secondary ions (e.g., C15N-) unique to streptavidin enabled unambiguous spatial mapping of the micropatterned protein with a lateral resolution of a few micrometers. TOF-SIMS imaging of micropatterned 15N-labeled streptavidin also illustrated the exquisite sensitivity of TOF-SIMS to low fractional coverage of protein (5 A effective thickness) in the background regions of the protein micropattern.

  20. 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.

  1. A minichaperone-based fusion system for producing insoluble proteins in soluble stable forms.

    Science.gov (United States)

    Sharapova, Olga A; Yurkova, Maria S; Fedorov, Alexey N

    2016-02-01

    We have developed a fusion system for reliable production of insoluble hydrophobic proteins in soluble stable forms. A carrier is thermophilic minichaperone, GroEL apical domain (GrAD), a 15 kDa monomer able to bind diverse protein substrates. The Met-less variant of GrAD has been made for further convenient use of Met-specific CNBr chemical cleavage, if desired. The Met-less GrAD retained stability and solubility of the original protein. Target polypeptides can be fused to either C-terminus or N-terminus of GrAD. The system has been tested with two unrelated insoluble proteins fused to the C-terminus of GrAD. One of the proteins was also fused to GrAD N-terminus. The fusions formed inclusion bodies at 25°C and above and were partly soluble only at lower expression temperatures. Most importantly, however, after denaturation in urea, all fusions without exception were completely renatured in soluble stable forms that safely survived freezing-thawing as well as lyophilization. All fusions for both tested target proteins retained solubility at high concentrations for days. Functional analysis revealed that a target protein may retain functionality in the fusion. Convenience features include potential thermostability of GrAD fusions, capacity for chemical and enzymatic cleavage of a target and His6 tag for purification. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  2. 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

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

    Energy Technology Data Exchange (ETDEWEB)

    Rastogi, Lori; Kora, Aruna Jyothi; Arunachalam, J., E-mail: aruncccm@gmail.com

    2012-08-01

    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: Black-Right-Pointing-Pointer Method for NaBH{sub 4} reduced and BSA capped gold nanoparticle was standardized. Black-Right-Pointing-Pointer Nanoparticles were spherical and nearly monodispersed with a size of 7.8 nm. Black-Right-Pointing-Pointer Nanoparticles are extremely stable towards pH modification and electrolyte addition. Black

  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. 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.

  6. Heavy metal concentration in coat hair and hoof horn in stabled and reserve Polish Konik horses.

    Science.gov (United States)

    Stachurska, A; Wałkuska, G; Chałabis-Mazurek, A; Jaworski, Z; Cebera, M

    2009-01-01

    The objective of the study was to determine if there were differences in Cr, Mn, Cu, Cd, Pb and Ni levels in coat hair and hoof horn between Polish Konik horses maintained in stabled and reserve systems in Roztocze National Park in Poland. Summer and winter feeding seasons (seasons of the year in case of hooves) and age of horses were taken into account. 107 samples of hair and hooves, as well as all kinds of feed and sources of water available for the horses were examined. The results of the study indicate that the concentration of the elements in the coat hair, hoof horn, feeds and water is usually too low to show possible influence of the system of maintenance in feeding seasons and age effect. The reserve horses are not more exposed to heavy metal contamination than the stabled horses. In the hair, solely Mn level is affected by the systems of maintenance in summer and winter feeding seasons: during winter it is higher, particularly in the stabled system, than during summer. In hooves, Cr concentration is higher in stabled horses than in reserve horses and conversely, Cu content is higher in the reserve horses. Mn content is age-affected both in hair and hooves, likewise is Cu content in the hooves. The results indicate a necessity of Cu compound supplementation in feeding the horses in both systems of maintenance.

  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. Cow hair allergen concentrations in dairy farms with automatic and conventional milking systems: From stable to bedroom.

    Science.gov (United States)

    Böhlandt, A; Schierl, R; Heizinger, J; Dietrich-Gümperlein, G; Zahradnik, E; Bruckmaier, L; Sültz, J; Raulf, M; Nowak, D

    2016-01-01

    Bovine hair and dander are considered to be a notable risk factor for sensitization and allergic symptoms in occupationally exposed cattle farmers due to various IgE binding proteins. Farmers are suspected not only to be exposed during their work inside the stables but also inside their homes as allergens could be transferred via hair and clothes resulting in continued bovine allergen exposure in private areas. In recent years a new sensitive sandwich ELISA (enzyme linked immunosorbent assay) test has been developed to measure the cow hair allergen (CHA) concentration in dust. The aim of the present study was to determine the CHA concentration in airborne and settled dust samples in stables and private rooms of dairy cattle farms with automatic milking systems (AM) and conventional milking systems (CM), also with respect to questionnaire data on farming characteristics. For this purpose different sampling techniques were applied, and results and practicability of the techniques were compared. Dust sampling was performed in the stable, computer room (only AM), changing room, living room and bedroom (mattress) of 12 dairy farms with automatic milking systems (AM group) and eight dairy farms with conventional milking systems (CM group). Altogether, 90 samples were taken by ALK filter dust collectors from all locations, while 32 samples were collected by an ion charging device (ICD) and 24 samples by an electronic dust fall collector (EDC) in computer rooms (AM) and/or changing and living rooms (not stables). The dust samples were extracted and analyzed for CHA content with a sandwich ELISA. At all investigated locations, CHA concentrations were above the limit of detection (LOD) of 0.1 ng/ml dust extract. The median CHA concentrations in dust collected by ALK filters ranged from 63 to 7154 μg/g dust in AM farms and from 121 to 5627 μg/g dust in CM farms with a steep concentration gradient from stables to bedrooms. ICD sampling revealed median CHA contents of 112

  9. 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.

  10. 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

  11. Present possibilities of protein concentrates production from plant sources

    OpenAIRE

    KRISTINOVÁ, Helena

    2011-01-01

    The aim of the bachelor thesis was gather information about present possibilities of proteins concentrates production from plant sources. I focused on dried pea (Pisum sativum L.), soybean (Glycine soja) and potato tuber (Solanum tuberosum L.). The protein concentrates from leguminous plants are produced by three basic processes, i.e. acid leasing (at ~pH 4.5), extracting with aqueous alcohol (60 - 90%), and denaturing the protein with moist heat before extraction with water. Pea protein is p...

  12. Engineering Environmentally-Stable Proteases to Specifically Neutralize Protein Toxins

    Science.gov (United States)

    2013-10-01

    synthesized as a fusion protein on the surface of M13 phage . The random library of subtilisin phage is mixed with the GA- PLFRAL-S-GB substrate...factor (LF), representing four functionally distinct families of toxins. The centerpiece of our design effort is a phage -display selection method for...hour assay. 15. SUBJECT TERMS Enterotoxin, protease, directed evolution, subtilisin, protein engineering, phage -display, enzymology 16. SECURITY

  13. Stable isotope and high precision concentration measurements confirm that all humans produce and exhale methane.

    Science.gov (United States)

    Keppler, Frank; Schiller, Amanda; Ehehalt, Robert; Greule, Markus; Hartmann, Jan; Polag, Daniela

    2016-01-29

    Mammalian formation of methane (methanogenesis) is widely considered to occur exclusively by anaerobic microbial activity in the gastrointestinal tract. Approximately one third of humans, depending on colonization of the gut by methanogenic archaea, are considered methane producers based on the classification terminology of high and low emitters. In this study laser absorption spectroscopy was used to precisely measure concentrations and stable carbon isotope signatures of exhaled methane in breath samples from 112 volunteers with an age range from 1 to 80 years. Here we provide analytical evidence that volunteers exhaled methane levels were significantly above background (inhaled) air. Furthermore, stable carbon isotope values of the exhaled methane unambiguously confirmed that this gas was produced by all of the human subjects studied. Based on the emission and stable carbon isotope patterns of various age groups we hypothesize that next to microbial sources in the gastrointestinal tracts there might be other, as yet unidentified, processes involved in methane formation supporting the idea that humans might also produce methane endogenously in cells. Finally we suggest that stable isotope measurements of volatile organic compounds such as methane might become a useful tool in future medical research diagnostic programs.

  14. 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)

  15. 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)

  16. 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.

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

    African Journals Online (AJOL)

    2018-04-04

    Apr 4, 2018 ... concentrate in a patient with mild‑to‑moderate dyspnea caused by swelling of the upper airway (larynx) and tongue. Keywords: C1 esterase inhibitor protein, hereditary angioedema, laryngeal edema, oropharyngeal swelling. Usefulness of C1 Esterase Inhibitor Protein Concentrate in the. Management of ...

  18. 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 ...

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

    Energy Technology Data Exchange (ETDEWEB)

    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 /sup 40/K, /sup 60/Co, /sup 95/Zr-Nb, /sup 106/Ru, /sup 137/Cs, /sup 226/Ra, and /sup 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.

  1. 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.

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

    African Journals Online (AJOL)

    Soy protein concentrate (SPC) and soy protein isolate (SPI) were limited hydrolyzed with trypsin or neutrase under controlled hydrolysis conditions. Eight soybean protein hydrolysates, 4 SPC hydrolysates and 4 SPI hydrolysates were prepared with degree of hydrolysis (DH) of 1 or 2%. SDS-PAGE was used to determine ...

  3. Methods to Concentrate Proteins for Protein Isolation, Proteomic, and Peptidomic Evaluation.

    Science.gov (United States)

    Goldring, J P Dean

    2015-01-01

    In protein isolation, proteomic, or peptidomic procedures, protein solutions are often concentrated to remove solvents and undesirable molecules, to separate protein fractions or to increase protein concentrations. Proteins can be concentrated by precipitation from solution with ammonium sulfate, polyethylene glycol, organic solvent, trichloroacetic acid, potassium chloride/sodium dodecyl sulfate, and three-phase partitioning. Solvents can be removed by passage through a semipermeable barrier where protein solutions are forced against the barrier in a centrifuge tube or with increased pressure concentrating protein in the remaining solution. The semipermeable barrier can be surrounded by a hygroscopic reagent to draw the solvent across the membrane. Proteins can be concentrated by freeze-drying (lyophilization). All these methods to concentrate proteins are discussed.

  4. 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...

  5. 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....

  6. New trends in bio/nanotechnology: stable proteins as advanced molecular tools for health and environment.

    Science.gov (United States)

    Staiano, M; Baldassarre, M; Esposito, M; Apicella, E; Vitale, R; Aurilia, V; D'Auria, S

    2010-01-01

    In this work the thermophilic trehalose/maltose-binding protein from Thermococcus litoralis is presented as a probe for the design of a high stable fluorescence biosensor for glucose. In particular, we show the possibility of modulating the protein specificity by changing temperature. In addition to glucose sensing, we also report on the possibility of utilizing odorant-binding proteins as a probe for the development of optical sensors for analytes of environmental interests.

  7. 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 (water isotopic values reflecting precipitation, indicating preferential flow (>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.

  8. 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.

  9. 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)

  10. 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.

  11. Nutritional quality of sunflower protein concentrate and isolate

    International Nuclear Information System (INIS)

    Niazi, A.H.K.; Kausar, T.; Ishaque, W.

    1994-01-01

    Sunflower Protein Concentrate (SPFC) prepared during the investigations contained 64.75 protein, 6.5% crude fibre and 6.15 phytic acid whereas crude protein, crude fibre and phytic acid in sunflower protein isolate were found to e 85.2% 4% and 2.5% respectively. Nutritional value i.e. per (2.17 and 2.35), NPR (3.10 and 3.32) and TD (80.5 and 82.2) of SFPC and SFPI showed their great potential as cheaper protein rich source of incorporation in food and feed. (author)

  12. Protein concentrates from defatted rice bran: preparation and characterization

    Directory of Open Access Journals (Sweden)

    Inajara Beatriz Brose PIOTROWICZ

    2017-10-01

    Full Text Available Abstract The goal of this work is to determine the optimal conditions for the obtaining of protein concentrates from rice bran. The effects of defatting processes applied to this product in laboratory and in industrial scales were investigated. Through two experimental designs were performed, and production conditions to obtain the protein concentrates were chosen with values of protein contents average 59.9% and 57.1%, with protein yields of 61.6% and 30.7%. The concentrates produced with industrially defatted rice bran showed higher digestibility and increased thermal stability compared with the product obtained with laboratory defatted rice bran. Both concentrates show molecular weight proteins below 50 kDa. The morphology of the precipitated proteins, analyzed by scanning electron microscopy, there was a great difference in the size of particles, which form the wet precipitates. The differences presented by the concentrates can be due to defatting processes which raw materials were submitted. Thus, studying the protein extraction conditions and knowing its characteristics is very important for the industry, because food processing requires knowing the behavior of each compound during and after processes they will be submitted.

  13. Freeze concentration of proteins in Antarctic krill wash water

    Science.gov (United States)

    Qi, Xiangming; Xu, Jing; Zhao, Kuo; Guo, Hui; Ma, Lei

    2017-12-01

    Water-washing removes fluoride from Antarctic krill but produces large volumes of wash water containing water- soluble proteins and fluoride. The freeze concentration method was tested to determine if it could be used to recover water-soluble proteins while leaving the fluoride in solution. After freezing and thawing the wash water, protein and fluoride contents of the thawed fractions were determined to explore the melting regularity of components in the wash water. The highest concentration factors of protein and fluoride were obtained after 80 min of thawing, such as 1.48 ± 0.06 and 1.35 ± 0.04 times, respectively. The free amino-nitrogen (FAN) content and sodium dodecyl sulfate-polyacrylamide gel electrophoresis pattern results revealed that the highest concentrations of all ingredients were obtained after 80 min of the process. The degree of hydrolysis of all fractions from the thawing process fluctuated in a narrow range around 12% during the entire process, indicating that the thawing order did not change with various proteins or time during the entire thawing course. These results demonstrate that the freeze concentration method can be used to concentrate protein solutions, even those with fluoride. It was concluded that condensation was achieved and no ingredient could be separated, regardless of fluoride, amino acids, or different proteins in the water.

  14. 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.

  15. QTL identification of grain protein concentration and its genetic ...

    Indian Academy of Sciences (India)

    concentration and grain weight using two population in maize (Zea mays L). J. Genet. 88, 61–67]. Introduction. Protein is one of the three most important storage chemi- cal components in maize grains. Generally, protein concen- tration (PC) is negatively and positively correlated with the other two grain components, starch ...

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

    African Journals Online (AJOL)

    2018-04-04

    Apr 4, 2018 ... of this case report is to describe the lifesaving use of a novel C1‑INH protein concentrate in a patient with mild‑to‑moderate dyspnea caused by swelling of the upper airway (larynx) and tongue. Keywords: C1 esterase inhibitor protein, hereditary angioedema, laryngeal edema, oropharyngeal swelling.

  17. 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...

  18. 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 ...

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

    African Journals Online (AJOL)

    The study evaluated methods of producing protein concentrates and isolates from cashew nut in order to increase the global utilization of cashew nut and increase the availability of affordable, good quality protein for people. Cashew nuts were processed to obtain roasted cashew kernel. The roasted cashew kernels were ...

  20. 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

  1. 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.

  2. Preparation of iron bound succinylated milk protein concentrate and evaluation of its stability.

    Science.gov (United States)

    Shilpashree, B G; Arora, Sumit; Sharma, Vivek; Bajaj, Rajesh Kumar; Tomar, S K

    2016-04-01

    Major problems associated with the fortification of soluble iron salts include chemical reactivity and incompatibility with other components. Milk protein concentrate (MPC) are able to bind significant amount of iron due to the presence of both casein and whey protein. MPC in its native state possess very poor solubility, therefore, succinylated derivatives of MPC (succ. MPC) were also used for the preparation of protein-iron complex. Preparation of the complex involved centrifugation (to remove insoluble iron), ultrafiltration (to remove unbound iron) and lyophilisation (to attain in dry form). Iron binding ability of MPC enhanced significantly (Piron from both varieties of complexes was monitored under different conditions encountered during processing. Higher stability (Piron was observed in succ. MPC-iron complex than native protein complex. This method could be adopted for the production of stable iron enriched protein, an organic iron source. Copyright © 2015 Elsevier Ltd. All rights reserved.

  3. 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

  4. 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.

  5. 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.

  6. Protein intake induced an increase in exercise stimulated fat oxidation during stable body weight.

    Science.gov (United States)

    Soenen, Stijn; Plasqui, Guy; Smeets, Astrid J; Westerterp-Plantenga, Margriet S

    2010-12-02

    Protein-rich weight-loss diets spare fat-free mass at the cost of fat mass. The objective was to examine if there is a change in stimulated fat oxidation related to protein intake during stable body weight. Subjects' (BMI 22±2kg/m(2), age 25±8 years) maximal fat oxidation (Fat(max)) was assessed during a graded bicycle test, before and after a 3-month dietary-intervention of 2MJ/day supplements exchanged with 2MJ/d of habitual energy intake. The parallel design consisted of protein-rich supplements in the protein group and an isocaloric combination of carbohydrate and fat supplements in the control group. Daily protein intake was determined according to 24-h urine nitrogen. Body composition was measured according to a 4-compartment model by a combination of underwater-weighing technique, deuterium-dilution technique and whole-body dual-energy X-ray absorptiometry (DXA). Subjects were weight stable and did not change their physical activity. The protein group (n=12) increased protein intake (11±14g, Pbody weight (Pbody composition or VO(2)max. Increased stimulated fat oxidation was related to increased protein intake. Copyright © 2010 Elsevier Inc. All rights reserved.

  7. Effect of raised plasma β endorphin concentrations on peripheral pain and angina thresholds in patients with stable angina

    OpenAIRE

    Jarmukli, N; Ahn, J; Iranmanesh, A; Russell, D

    1999-01-01

    OBJECTIVE—To determine whether changes in plasma concentrations of β endorphins alter angina threshold and peripheral pain threshold in patients with stable angina.
DESIGN—Latin square design comparison of angina thresholds by exercise treadmill test and peripheral pain thresholds using a radiant heat source in eight patients with stable angina under control conditions, after stimulation of pituitary β endorphin release by ketoconazole, after suppression of pituitary β endorphin release by de...

  8. 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.

  9. Peritoneal dialysate effluent and serum CA125 concentrations in stable peritoneal dialysis patients.

    Science.gov (United States)

    Redahan, Lynn; Davenport, Andrew

    2016-06-01

    CA125 in peritoneal dialysis (PD) effluent dialysate has been used as a surrogate biomarker for the health of the peritoneum in PD patients. However CA125 is synthesised by epithelial cells and as such is not specific for the peritoneum, and most studies have only measured peritoneal CA125, without serum CA125 values. As such we wished to determine the factors which influenced PD effluent CA125 in a large contemporaneous cohort. We measured dialysate effluent CA125 in PD patients attending for routine assessment of peritoneal membrane function with a peritoneal equilibration test (PET), with corresponding serum CA125. Serum and dialysate CA125 were measured in 205 PD patients; 59.0 ± 16.8 years, median PD treatment 3 (2-20) months, 59 % male, 42.4 % diabetic, with 31.2 % treated by continuous ambulatory peritoneal dialysis, 22 % by automated overnight peritoneal dialysis cycler (APD) and 46.8 % by APD with a day time exchange. The median serum CA125 was 21 (13-38) U/ml, with an effluent 4 h PD PET effluent of 20 (11.5-36.5) U/ml. PET PD effluent dialysate was associated with PET dialysate total protein (β 12.9, p peritonitis episodes. PD effluent CA125 concentrations were associated with peritoneal protein losses and increased by the usage of higher glucose dialysates to compensate for loss of residual renal function.

  10. Texture profile in processed cheese: influence of the use of milk protein concentrates and whey protein concentrates

    Directory of Open Access Journals (Sweden)

    Alisson Borges Souza

    2014-07-01

    Full Text Available The techno-functional properties of proteins related with the molecular characteristics are facilitated by partial unfolding of structures. From these interactions, the medium pH is presented as a major interferer in intensity and type of reaction that takes place. The intensity of denaturation and interaction of different proteins occur in different forms and intensity accordingly to the pH value of the medium in which they are located. This study aimed to verify the influence of interactions between whey protein concentrate/milk protein concentrate on the evolution of the texture profile of processed cheese at different pH values. We have analyzed samples of commercial whey protein concentrate (WPC and milk protein concentrate (MPC using 112.5g/kg processed cheese. The results were interpreted in terms of texture profile. It was also possible to optimize the different proportions of WPC and MPC, and pH value change the parameters of texture for creamy processed cheese and the pH was also an influencing factor in this optimization.

  11. 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.

  12. 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.

  13. 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.

  14. An immunoglobulin binding protein (antigen 5) of the stable fly (Diptera: Muscidae) salivary gland stimulates bovine immune responses.

    Science.gov (United States)

    Ameri, M; Wang, X; Wilkerson, M J; Kanost, M R; Broce, A B

    2008-01-01

    The stable fly, Stomoxys calcitrans (L.) (Diptera: Muscidae), is an economically important pest of livestock. Previous studies demonstrated lymphocyte suppression by crude salivary gland extract (SGE) of the stable fly. A dominant 27-kDa protein identified in the SGE was reported to stimulate immunodominant antibody responses in exposed cattle. The purpose of this study was to determine whether this protein, now identified as ahomolog of insect proteins named antigen 5 (Ag5), was responsible for the lymphocyte suppression and whether naive calves can mount an immune response to it. Calves raised in the winter were immunized with recombinant Ag5 (rAg5) expressed in Drosophila S2 cells or with "natural" Ag5 protein isolated by preparative gel electrophoresis of SGE. Control calves were immunized with adjuvant alone. Rising antibody concentrations to rAg5 were detected in two of three calves immunized with rAg5 and one of three calves immunized with natural Ag5. Recall lymphocyte responses to rAg5 were detected at 21 and 28 d postimmunization in calves immunized with rAg5 but not in calves immunized with the natural Ag5 or those exposed to adjuvant alone. Mitogen-stimulated bovine lymphocyte responses were not suppressed by rAg5. Further investigation using immunoblotting revealed that rAg5 binds to the Fc and F (ab')2 portions of bovine IgG, but not to an Fab fragment. These findings suggest that Ag5 of the stable fly salivary gland is not immunosuppressive but that it has immunoglobulin binding properties and can invoke specific antibody and memory lymphocyte responses in immunized calves.

  15. Stable accumulation of seed storage proteins containing vaccine peptides in transgenic soybean seeds.

    Science.gov (United States)

    Maruyama, Nobuyuki; Fujiwara, Keigo; Yokoyama, Kazunori; Cabanos, Cerrone; Hasegawa, Hisakazu; Takagi, Kyoko; Nishizawa, Keito; Uki, Yuriko; Kawarabayashi, Takeshi; Shouji, Mikio; Ishimoto, Masao; Terakawa, Teruhiko

    2014-10-01

    There has been a significant increase in the use of transgenic plants for the large-scale production of pharmaceuticals and industrial proteins. Here, we report the stable accumulation of seed storage proteins containing disease vaccine peptides in transgenic soybean seeds. To synthesize vaccine peptides in soybean seeds, we used seed storage proteins as a carrier and a soybean breeding line lacking major seed storage proteins as a host. Vaccine peptides were inserted into the flexible disordered regions in the A1aB1b subunit three-dimensional structure. The A1aB1b subunit containing vaccine peptides in the disordered regions were sorted to the protein storage vacuoles where vaccine peptides are partially cleaved by proteases. In contrast, the endoplasmic reticulum (ER)-retention type of the A1aB1b subunit containing vaccine peptides accumulated in compartments that originated from the ER as an intact pro-form. These results indicate that the ER may be an organelle suitable for the stable accumulation of bioactive peptides using seed storage proteins as carriers. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  16. 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...

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

    African Journals Online (AJOL)

    STORAGESEVER

    2008-08-04

    Aug 4, 2008 ... Fractionation led to increase in the gross energy by 22.0% and decrease in the phytate and tannin contents ..... SBM = Soybean meal, LPC = Telfairia occidentalis leaf protein concentrate, SBM = Soybean meal, PCV= Packed cell volume, WBC = White blood cell, RBC = Redblood cell, Hbc = Haemoglobin.

  18. Free proline, soluble sugars and soluble proteins concentration as ...

    African Journals Online (AJOL)

    In this study, the effects of salt stress on free proline, soluble sugars and soluble proteins accumulation were investigated in two sugarcane (Saccharum sp.) cultivars: CP66-346 (salt- tolerant) and CP65-357 (salt-sensitive). Young plants of these cultivars were exposed, in a hydroponic system, to four concentrations of NaCl ...

  19. QTL identification of grain protein concentration and its genetic ...

    Indian Academy of Sciences (India)

    Therefore, it is desirable to conduct further studies using more relevant germplasm to reveal the genetic nature of PC. QTL map- ping using several connected multi-parental crosses could in-. Keywords. high-oil maize; protein concentration; F2:3 family lines; single-trait and joint-trait QTL analysis; genetic backgrounds.

  20. Comparison of different protein concentration techniques within preformulation development.

    Science.gov (United States)

    Eppler, Annette; Weigandt, Markus; Schulze, Sandra; Hanefeld, Andrea; Bunjes, Heike

    2011-12-12

    Highly concentrated antibody solutions are of increasing importance in the pharmaceutical industry. During production highly concentrated solutions are usually prepared by tangential flow filtration (TFF). Since this technique is often not applicable in the early phase of formulation development, where the available amounts of protein are commonly very small, small scale techniques like dialysis or ultrafiltration with stirred cells or centrifugal filters have to be employed. In this study the small scale techniques were compared to tangential flow filtration, with regard to the quality and stability of the concentrated products. The achievable concentration of a protein, when starting from a model antibody solution with 10mg/ml, was also assessed. Concentrations above 100mg/ml could be obtained with all techniques, however with different product qualities. The stability of the highly concentrated solutions (100 mg/ml) was analyzed by turbidity measurements, size exclusion chromatography (SEC), SDS-PAGE and isoelectric focusing (IEF) after storage at 25 and 40°C for 8 weeks. Solutions prepared by dialysis exhibited the smallest degree of instability, whereas those manufactured by centrifugal filtration revealed the best comparability to products obtained by tangential flow filtration with regard to the results of isoelectric focusing, turbidity measurements (UV-vis) and size exclusion chromatography. Stability differences were observed within all analytical methods, primarily after storage and not directly after the concentration process. Copyright © 2011 Elsevier B.V. All rights reserved.

  1. 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

  2. Adverse pregnancy outcomes in hypertensive patients: predictive value of protein concentration versus total protein.

    Science.gov (United States)

    Lee, Amy M; Briandet, Benjamin M; Caranta, Diane G; Zelig, Craig M

    2014-11-01

    To compare the predictive value of protein concentration in a twenty-four hour urine collection to the conventional total protein in a twenty-four hour urine collection for adverse pregnancy outcomes in hypertensive patients. Retrospective cohort study. Hypertensive patients ≥20 weeks estimated gestational age (EGA) who completed twenty-four hour urine protein collections were identified; antepartum and delivery data were examined. For study patients who met criteria for adverse pregnancy outcome, multi-variable analysis was performed and summary receiver operating characteristic (ROC) curves were generated for each model (total protein compared to protein concentration). The models were compared by analyzing the area under the curve (AUC). A total of 150 patients were analyzed. Mean gestational age at delivery was 36.7 weeks. Analysis of the ROC curves showed no significant difference between the models (AUCs of 0.668 versus 0.656, p = 0.715). Optimal thresholds were 299.2 mg for total protein and 0.1 mg/ml for protein concentration. A protein concentration of 0.1 mg/ml on a twenty-four hour urine collection appears equivalent to the traditional 300 mg total protein. If confirmed by prospective studies, this finding would be clinically important in cases where collections fall short of the 300 mg threshold but exceed the 0.1 mg/ml concentration.

  3. 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

  4. G protein-coupled receptors form stable complexes with inwardly rectifying potassium channels and adenylyl cyclase.

    Science.gov (United States)

    Lavine, Natalie; Ethier, Nathalie; Oak, James N; Pei, Lin; Liu, Fang; Trieu, Phan; Rebois, R Victor; Bouvier, Michel; Hebert, Terence E; Van Tol, Hubert H M

    2002-11-29

    A large number of studies have demonstrated co-purification or co-immunoprecipitation of receptors with G proteins. We have begun to look for the presence of effector molecules in these receptor complexes. Co-expression of different channel and receptor permutations in COS-7 and HEK 293 cells in combination with co-immunoprecipitation experiments established that the dopamine D(2) and D(4), and beta(2)-adrenergic receptors (beta(2)-AR) form stable complexes with Kir3 channels. The D(4)/Kir3 and D(2) receptor/Kir3 interaction does not occur when the channel and receptor are expressed separately and mixed prior to immunoprecipitation, indicating that the interaction is not an artifact of the experimental protocol and reflects a biosynthetic event. The observed complexes are stable in that they are not disrupted by receptor activation or modulation of G protein alpha subunit function. However, using a peptide that binds Gbetagamma (betaARKct), we show that Gbetagamma is critical for dopamine receptor-Kir3 complex formation, but not for maintenance of the complex. We also provide evidence that Kir3 channels and another effector, adenylyl cyclase, are stably associated with the beta(2)-adrenergic receptor and can be co-immunoprecipitated by anti-receptor antibodies. Using bioluminescence resonance energy transfer, we have shown that in living cells under physiological conditions, beta(2)AR interacts directly with Kir3.1/3.4 and Kir3.1/3.2c heterotetramers as well as with adenylyl cyclase. All of these interactions are stable in the presence of receptor agonists, suggesting that these signaling complexes persist during signal transduction. In addition, we provide evidence that the receptor-effector complexes are also found in vivo. The observation that several G protein-coupled receptors form stable complexes with their effectors suggests that this arrangement might be a general feature of G protein-coupled signal transduction.

  5. 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....

  6. 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

  7. Concentration Dependent Ion-Protein Interaction Patterns Underlying Protein Oligomerization Behaviours

    Science.gov (United States)

    Batoulis, Helena; Schmidt, Thomas H.; Weber, Pascal; Schloetel, Jan-Gero; Kandt, Christian; Lang, Thorsten

    2016-04-01

    Salts and proteins comprise two of the basic molecular components of biological materials. Kosmotropic/chaotropic co-solvation and matching ion water affinities explain basic ionic effects on protein aggregation observed in simple solutions. However, it is unclear how these theories apply to proteins in complex biological environments and what the underlying ionic binding patterns are. Using the positive ion Ca2+ and the negatively charged membrane protein SNAP25, we studied ion effects on protein oligomerization in solution, in native membranes and in molecular dynamics (MD) simulations. We find that concentration-dependent ion-induced protein oligomerization is a fundamental chemico-physical principle applying not only to soluble but also to membrane-anchored proteins in their native environment. Oligomerization is driven by the interaction of Ca2+ ions with the carboxylate groups of aspartate and glutamate. From low up to middle concentrations, salt bridges between Ca2+ ions and two or more protein residues lead to increasingly larger oligomers, while at high concentrations oligomers disperse due to overcharging effects. The insights provide a conceptual framework at the interface of physics, chemistry and biology to explain binding of ions to charged protein surfaces on an atomistic scale, as occurring during protein solubilisation, aggregation and oligomerization both in simple solutions and membrane systems.

  8. Tracking the Path of Carbon from Plants to Rhizobiomes using Protein Stable Isotope Probing

    Science.gov (United States)

    Lipton, M. S.; Nicora, C.; Shaw, J. B.; Karaoz, U.; Nuccio, E. E.; Zhalnina, K.; Paša-Tolić, L.; Brodie, E.; Pett-Ridge, J.; Firestone, M.

    2016-12-01

    Plants and microorganisms have complex relationships that mediate terrestrial carbon fluxes. Understanding the functional roles of belowground biotic communities and their interactions with plant roots is critical to understanding the fate of plant carbon in soil. Currently many `omics-driven technologies reveal only a static snap shot of functionality in biological communities, but temporally-resolved tools are necessary describe biological processes that change and evolve over time. Stable isotope probing (SIP) experiments are temporal in nature, and can track the flow of carbon from the atmosphere through the plant and into the soil microbial community through space and time, and ultimately reveal both plant-microbe interactions and the fate of the carbon within the system. The combination of 'omics approaches with SIP allows us to selectively target and characterize metabolically active organisms in a community, and elucidate functional potential over time-scales relevant to biological processes. Protein stable isotope probing (Pro-SIP) offers a unique perspective into the interaction between plants and microbial communities, and potentially links taxonomic, functional and metabolic information. We developed a Pro-SIP approach amenable for use in soil systems using soils collected from an annual grassland in Northern CA amended with 13C labeled glucose. Using this approach we determined the overall proteomic profile and recently synthesized proteins within the plant-associated microbial community. The mass spectrometric analysis of the proteins extracted from the soil identified over 60,000 proteins that correspond to about 2200 protein functions. Of these functions, over 500 were found to contain a labeled peptide. Proteins related to glucose metabolism, carbohydrate storage, sugar transport and sulfur metabolism were found to be actively synthesized after glucose amendment. In systems where plants were grown with 13CO2 revealed synthesis of central metabolism

  9. The quality control of fruit juices by using the stable isotope ratios and trace metal elements concentrations

    Science.gov (United States)

    Magdas, D. A.; Dehelean, A.; Puscas, R.; Cristea, G.; Tusa, F.; Voica, C.

    2012-02-01

    In the last years, a growing number of research articles detailing the use of natural abundance light stable isotopes variations and trace metal elements concentration as geographic "tracers" to determine the provenance of food have been published. These investigations exploit the systematic global variations of stable hydrogen, oxygen and carbon isotope ratios in (combination) relation with trace metal element concentrations. The trace metal elements content of plants and also their light stable isotopic ratios are mainly related to the geological and pedoclimatic characteristics of the site of growth. The interpretation of such analysis requires an important number of data for authentic natural juices regarding the same seasonal and regional origin, because the isotopic analysis parameters of fruit juices show remarkable variability depending on climatologically factors. In this work was mesured H, C, O stable isotope ratios and the concentrations of 16 elements (P, K, Mg, Na, Ca, Cu, Cr, Ni, Zn, Pb, Co, As, Cd, Mn, Fe and Hg) from 12 single strength juices. The natural variations that appear due to different environmental and climatic conditions are presented and discussed.

  10. 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

  11. Functional Characteristics of Milk Protein Concentrates and Their Modification.

    Science.gov (United States)

    Uluko, Hankie; Liu, Lu; Lv, Jia-Ping; Zhang, Shu-Wen

    2016-05-18

    A major deterrent to the usage of milk protein concentrate (MPC), a high-protein milk product with increasing demand as a food and sports drink ingredient, has been its poor functional characteristics when compared with other milk protein products such as whey protein concentrate and sodium caseinates. This review discusses the recent research on functional properties of MPC, focusing on factors that may contribute to the poor functional characteristics before, during, and after production. Current research, methods employed, and new understanding on the causes of poor solubility of MPC at mild temperatures (about 20°C) has been presented, including loss of solubility during storage as these areas have received unprecedented attention over the past decade, and also affects other useful functional properties of MPC, such as emulsifying properties, gelation, and foaming. Processing methods, which include heat treatment, high-pressure application, microwave heating, ultrasound application, and enzyme and salts modification, have been used or have potential to modify or improve the functional properties of MPCs. Future research on the effects of these processing methods on the functional properties, including effects of enzyme hydrolysis on bitterness and bioactivity, has also been discussed.

  12. Dual Topology of the Melanocortin-2 Receptor Accessory Protein (MRAP Is Stable

    Directory of Open Access Journals (Sweden)

    Zachary J. Maben

    2016-07-01

    Full Text Available MRAP (melanocortin 2 receptor accessory protein facilitates trafficking of melanocortin 2 (MC2 receptors and is essential for ACTH binding and signaling. MRAP is a single transmembrane domain protein that forms antiparallel homodimers. These studies ask when MRAP first acquires this dual topology, whether MRAP architecture is static or stable, and whether the accessory protein undergoes rapid turnover. To answer these questions we developed a novel approach that capitalizes on the specificity of bacterial biotin ligase, which adds biotin to lysine in a short acceptor peptide sequence; the distinct mobility of MRAP protomers of opposite orientations based on their N-linked glycosylation; and the ease of identifying biotin-labeled proteins. We inserted biotin ligase acceptor peptides at the N- or C-terminal ends of MRAP and expressed the modified proteins in mammalian cells together with either cytoplasmic or endoplasmic reticulum-targeted biotin ligase. MRAP assumed dual topology early in biosynthesis in both CHO and OS3 adrenal cells. Once established, MRAP orientation was stable. Despite its conformational stability, MRAP displayed a half life of under 2 hr in CHO cells. The amount of MRAP was increased by the proteasome inhibitor MG132 and MRAP underwent ubiquitylation on lysine and other amino acids. Nonetheless, when protein synthesis was blocked with cycloheximide, MRAP was rapidly degraded even when MG132 was included and all lysines were replaced by arginines, implicating non-proteasomal degradation pathways. The results show that although MRAP does not change orientations during trafficking its synthesis and degradation are dynamically regulated.

  13. Direct molecular evolution of detergent-stable G protein-coupled receptors using polymer encapsulated cells.

    Science.gov (United States)

    Scott, Daniel J; Plückthun, Andreas

    2013-02-08

    G protein-coupled receptors (GPCRs) are the largest class of pharmaceutical protein targets, yet drug development is encumbered by a lack of information about their molecular structure and conformational dynamics. Most mechanistic and structural studies as well as in vitro drug screening with purified receptors require detergent solubilization of the GPCR, but typically, these proteins exhibit only low stability in detergent micelles. We have developed the first directed evolution method that allows the direct selection of GPCRs stable in a chosen detergent from libraries containing over 100 million individual variants. The crucial concept was to encapsulate single Escherichia coli cells of a library, each expressing a different GPCR variant, to form detergent-resistant, semipermeable nano-containers. Unlike naked cells, these containers are not dissolved by detergents, allowing us to solubilize the GPCR proteins in situ while maintaining an association with the protein's genetic information, a prerequisite for directed evolution. The pore size was controlled to permit GPCR ligands to permeate but the solubilized receptor to remain within the nanocapsules. Fluorescently labeled ligands were used to bind to those GPCR variants inside the nano-containers that remained active in the detergent tested. With the use of fluorescence-activated cell sorting, detergent-stable mutants derived from two different family A GPCRs could be identified, some with the highest stability reported in short-chain detergents. In principle, this method (named cellular high-throughput encapsulation, solubilization and screening) is not limited to engineering stabilized GPCRs but could be used to stabilize other proteins for biochemical and structural studies. Copyright © 2012. Published by Elsevier Ltd.

  14. Synergistic effect of temperature, protein and salt concentration on structures and interactions among lysozyme proteins

    Science.gov (United States)

    Kundu, Sarathi; Aswal, V. K.; Kohlbrecher, Joachim

    2016-07-01

    Synergistic effect of temperature, protein and salt concentration on structures and interactions among lysozyme proteins in solution has been studied using small angle neutron scattering technique. Scattering study shows that for a particular protein concentration, with increasing temperature, short-range attraction decreases but long-range repulsion becomes system specific. In absence of salt, lower value of attractive interaction is obtained, however, in presence of salt it becomes higher and decreases with increasing temperature. For specific condition, weak long range attraction and intermediate range repulsion exists. At higher temperature (90 °C), fractal structure develops and the corresponding fractal dimension depends upon the experimental conditions.

  15. 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

  16. Rheological properties of whey proteins concentrate before and

    Directory of Open Access Journals (Sweden)

    Zoran Herceg

    2001-04-01

    Full Text Available Hydrocolloids are long-chain polymers, used in food production at small quantities (from 0,05 to 5 % to achieve appropriate rheological properties, prevent syneresis, increase the viscosity and stability of foodstuffs and for crystallization process control. The aim of this work was to investigate the influence of tribomechanical micronization of powdered whey protein concentrate on the rheological properties of whey proteins model systems as well as the influence of severalcarboxymethylcellulose hydrocolloids addition in such systems.Measurements were done using rotational viscosimeter, Brookfield DV-III at temperature 20 oC. The rheological parameters were determined by powerlaw model. The results of investigation have shown that all investigated systems are non-Newtonian. Depending on the pretreatment of whey proteins and the mass fractions of hydrocolloids they exhibited pseudoplastic or dilatant properties.Particle size analysis was performed using Fritsch – laser particle sizer “analysette 22”. The operation of tribomechanical micronization caused the decreasing of particle size and incrasing specific area of whey proteins. Tribomechanical treatment of whey proteins had significant influence on the rheological parameters and the type of flowing.

  17. Exopolysaccharides modify functional properties of whey protein concentrate.

    Science.gov (United States)

    Deep, G; Hassan, A N; Metzger, L

    2012-11-01

    The objective of this research was to produce whey protein concentrate (WPC) with modified functionality using exopolysaccharide- (EPS) producing cultures. Two different EPS-producing cultures, Lactococcus lactis ssp. cremoris JFR and Streptococcus thermophilus, producing EPS1 and EPS2 respectively, were used in this study. One EPS-nonproducing commercial cheese culture (DVS 850; Chr. Hansen, Milwaukee, WI) was used as the control. Reconstituted sweet whey powder was used in this study to eliminate variations from fresh whey. Cultures grown overnight in reconstituted WPC (10% wt/vol) were added, directly or after overnight cooling (cooled EPS), at 2% (wt/vol) to 6% (wt/wt) solution of reconstituted whey. Whey was then high-temperature, short-time pasteurized at 75 °C for 35s and ultrafiltered to a volume reduction factor of 5. Ultrafiltered whey (retentate) was spray dried at inlet and outlet air temperatures of 200 and 90 °C, respectively, to obtain WPC. In general, the solubility of WPC was higher at pH 7 than at pH 3. Whey protein concentrate containing EPS2 exhibited higher protein solubility than did WPC containing no EPS. Also, the presence of EPS in WPC decreased protein denaturation. The emulsifying ability of WPC containing EPS was higher than that in control. Addition of EPS to WPC significantly enhanced its gelling ability. Foam overrun and hydrophobicity of WPC were not affected by addition of EPS. In conclusion, data obtained from this study show that EPS modify WPC functionality. The extent of modification depends on the type of EPS. Cooling of culture containing EPS before its addition to whey further reduced WPC protein denaturation and increased its solubility at pH 7 and gel hardness. Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  18. 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 <2%. Concerning diet, foals fed high protein spent more time in movement (19.62±0.73% of day v. 10.45±0.73% in low-protein (LP) foals; P⩽0.001), whereas the LP group increased resting (43.42±1.12% v. 38.02±1.12%; P⩽0.001). No stereotypies were found, and daily activity followed the typical values for 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.

  19. [Expression of human Jagged-1 protein on eukaryotic cells and establishment of stable transfectant cell line].

    Science.gov (United States)

    Gan, Zhi-Hua; Chen, Yu; Yan, Hua; Wang, Kan-Kan

    2010-08-01

    Jagged-1 protein is one of the ligands belonging to Notch signaling pathway. Notch signaling pathway is one of the major signaling pathways mediated by contact between cells and plays an important role to regulate the process of proliferation and differentiation of hematopoietic cells in the hematopoietic microenvironment. To study the biological effect after the combination of receptor and ligand in Notch signaling pathway and the mechanism of Notch signaling pathway in bone marrow stromal cells mediated-drug resistance, a NIH-3T3 cell line over-expressing Jagged-1 protein was constructed for further research purposes. A full coding region of Jagged-1 gene was cloned and inserted into eukaryotic expression plasmid to construct pEGFP-IRES2-Jagged-1 eukaryotic expression vector, then transfected into NIH-3T3 cell line, a mammalian cells. As a result Western blot analysis confirmed that the transfectant NIH-3T3 cells highly expressed Jagged-1 protein and flow cytometry analysis confirmed that the NIH-3T3-pEGFP-IRES2-Jagged-1 cell line over-expressed Jagged-1 protein was monoclonal after screened by selective medium and limiting dilution analysis. It is concluded that the pEGFP-IRES2-Jagged-1 eukaryotic expression vector and a stable transfectant monoclonal NIH-3T3 cell line are successfully established. The construction of the stable transfectant monoclonal NIH-3T3 cell line which overexpressed Jagged-1 protein, provides the conditions to further study the mechanism of the bone marrow stromal cell-mediated drug resistance and to discover the new drug targets.

  20. 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...... can be improved by pre-wilting to a higher DM concentration before ensiling. Abbreviations AA, amino acids; AA-N, amino acid nitrogen; ADL, acid detergent lignin; aNDFom, neutral detergent fibre assayed with heat stable amylase and expressed exclusive of residual ash; CP, crude protein; CH4, methane...

  1. 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.

  2. Fractionation, concentration and flow: A model coupling stable isotope ratios to fluid travel time and chemical reactivity

    Science.gov (United States)

    Druhan, J. L.; Maher, K.

    2014-12-01

    From the point of infiltration to the point of discharge, the chemical signature imparted to fluid flowing through catchments represents the weathering flux from the landscape. The magnitude of this flux is linked to both the time water spends in the system and the time required for reactions to influence fluid chemistry. The ratio of these characteristic times is often represented as a Damköhler number (Da), which links the parameters governing reactivity and flow. Stable isotope ratios are now commonly applied to identify and even quantify the processes and rates of primary mineral weathering, secondary mineral formation and biogeochemical cycling within catchments. Here, we derive a series of fractionation-discharge relationships for a variety of governing chemical rate laws utilizing Da coefficients. These equations can be used to isolate and quantify the effects of (1) fluid travel time distributions and (2) chemical weathering efficiency on observed stable isotope ratios. The analytical solutions are verified against multi-component reactive transport simulations of stable isotope fractionation in homogeneous and spatially correlated heterogeneous flow fields using the CrunchTope code and evaluated against field observations. We demonstrate that for an irreversible reaction, the relationship between stable isotope enrichment and reactant concentration obeys a Rayleigh-type model across a wide range of reaction rates. However, this relationship is violated when a heterogeneous travel time distribution is considered. This observation highlights an important discrepancy in the commonly assumed relationship between fractionation and concentration for irreversible reactions. We further extend our derivation to consider isotope fractionation associated with a reversible reaction (i.e. a kinetically controlled approach to equilibrium) in a steady-state flow field. Due to the dependence of the observed isotope ratio on the flow rate, kinetic enrichment and

  3. Mercury concentrations in Southern Beaufort Sea polar bears: variation based on stable isotopes of carbon and nitrogen.

    Science.gov (United States)

    Cardona-Marek, Tamara; Knott, Katrina K; Meyer, Benjamin E; O'Hara, Todd M

    2009-07-01

    Total Hg concentration was measured in hair and whole blood of 52 adult Southern Beaufort Sea polar bears (Ursus maritimus) captured in the spring of 2005. Stable isotopic signatures (i.e., 13C/12C, delta13C; 15N/14N, delta15N) in hair and two blood compartments (packed blood cells/clot and serum) were determined to assess the variation of Hg concentrations among polar bears in relation to their feeding ecology and other biological factors. Concentrations of Hg in hair and blood (2.2-23.9 microg/g dry wt and 0.007-0.213 microg/g wet wt, respectively) were within the range of values previously reported for polar bears in Canada and East Greenland. Mercury concentration in hair from females was higher than that in hair from males, and concentration was related to interactions between delta13C, delta15N, and longitude of capture location. Mercury concentrations in hair were inversely correlated to delta13C in hair and blood, suggesting that polar bears with greater total Hg concentrations fed more on pelagic prey, such as ringed seals or beluga whale, than on benthic prey. Variability in Hg concentrations in polar bear hair and blood may be the result of intraspecific or regional variation in prey selection rather than strictly trophic level interactions.

  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. Rational automatic search method for stable docking models of protein and ligand.

    Science.gov (United States)

    Mizutani, M Y; Tomioka, N; Itai, A

    1994-10-21

    An efficient automatic method has been developed for docking a ligand molecule to a protein molecule. The method can construct energetically favorable docking models, considering specific interactions between the two molecules and conformational flexibility in the ligand. In the first stage of docking, likely binding modes are searched and estimated effectively in terms of hydrogen bonds, together with conformations in part of the ligand structure that includes hydrogen bonding groups. After that part is placed in the protein cavity and is optimized, conformations in the remaining part are also examined systematically. Finally, several stable docking models are obtained after optimization of the position, orientation and conformation of the whole ligand molecule. In all the screening processes, the total potential energy including intra- and intermolecular interaction energy, consisting of van der Waals, electrostatic and hydrogen bonding energies, is used as the index. The characteristics of our docking method are high accuracy of the results, fully automatic generation of models and short computational time. The efficiency of the method was confirmed by four docking trials using two enzyme systems. In two attempts to dock methotrexate to dihydrofolate reductase and 2'-GMP to ribonuclease T1, the exact structures of complexes in crystals were reproduced as the most stable docking models, without any assumptions concerning the binding modes and ligand conformations. The most stable docking models of dihydrofolate and trimethoprim, respectively, to dihydrofolate reductase were also in good agreement with those suggested by experiment. In all test cases, it was shown that our method can accurately predict the correct docking structures, discriminating the correct model from incorrect ones. The efficiency of our method was further tested from the viewpoint of ability to predict the relative stability of the docking structures of two triazine derivatives to

  6. The efficacy of scale sampling for monitoring trace element concentrations and stable isotopes in commercially harvested walleye (Sander vitreus).

    Science.gov (United States)

    Ofukany, Amy F A; Hobson, Keith A; Wassenaar, Leonard I; Bond, Alexander L

    2015-01-01

    Commercial and sport fishes are subject to rigorous monitoring for concentrations of elements that could pose threats to human health, with numerous advisories issued by authorities annually for those fisheries with high mercury (Hg) concentrations. In Lake Winnipeg, Manitoba, Canada, the commercial walleye fishery is valued at more than $20 million/year, but has historically been subject to Hg advisories. We used an information theoretic approach to evaluate the utility of non-destructive fish-scale sampling to predict As, Mn and Hg concentrations, as well as stable isotope values in walleye muscle by analysing paired samples. Hg concentrations in scales were significantly related to those in muscle (r(2) = 0.75), but the relationships were weaker for As and Mn. The δ(15)N values in scales predicted δ(15)N in muscle reasonably well (r(2) = 0.72), whereas scale δ(13)C and δ(34)S had less predictive power for estimating their respective muscle stable isotope values. For all three isotope values, sex was a marginal predictor, with parameter confidence intervals bounding zero. Analytical constraints currently limit the utility of non-destructively analysing scales for Hg, but hindcasting trophic changes using archived walleye scales may be useful in understanding shifts in nutrients and production, particularly in impacted lake systems.

  7. Influence of Biopolym Granulat effects on reductionof ammonia concentration in stables of intensive farm animals breeding

    Directory of Open Access Journals (Sweden)

    Bohuslav Čermák

    2014-10-01

    Full Text Available The living environment distress is connected currently not only with industrial production but also agriculture is biggest producer of toxic gas – ammonia (NH3 .Emissions of that gas originate mainly in the farm animals breeding and generate within storage and handling with farmyard manure, slurry, poultry excrements and litter. Agriculture influences considerably landscape. has impact on basic effect on soil, water and air. In assessing experiment the preparation Biopolym Granulat rumen metabolism and N-balance was found positive effects in terms of increased ammonia nitrogen, the number of ciliates and the reduction of N-compounds in feces. Confirmed the impact on the ammonia content in well-ventilated dairy stable. The economic evaluation depends on the exercise price of milk.

  8. 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.

  9. Stable transformation of an episomal protein-tagging shuttle vector in the piscine diplomonad Spironucleus vortens

    Directory of Open Access Journals (Sweden)

    Cronembold Daniela

    2008-04-01

    Full Text Available Abstract Background Diplomonads are common free-living inhabitants of anoxic aquatic environments and are also found as intestinal commensals or parasites of a wide variety of animals. Spironucleus vortens is a putatively commensal diplomonad of angelfish that grows to high cell densities in axenic culture. Genomic sequencing of S. vortens is in progress, yet little information is available regarding molecular and cellular aspects of S. vortens biology beyond descriptive ultrastructural studies. To facilitate the development of S. vortens as an additional diplomonad experimental model, we have constructed and stably transformed an episomal plasmid containing an enhanced green fluorescent protein (GFP tag, an AU1 epitope tag, and a tandem affinity purification (TAP tag. This construct also contains selectable antibiotic resistance markers for both S. vortens and E. coli. Results Stable transformants of S. vortens grew relatively rapidly (within 7 days after electroporation and were maintained under puromycin selection for over 6 months. We expressed the enhanced GFP variant, eGFP, under transcriptional control of the S. vortens histone H3 promoter, and visually confirmed diffuse GFP expression in over 50% of transformants. Next, we generated a histone H3::GFP fusion using the S. vortens conventional histone H3 gene and its native promoter. This construct was also highly expressed in the majority of S. vortens transformants, in which the H3::GFP fusion localized to the chromatin in both nuclei. Finally, we used fluorescence in situ hybridization (FISH of the episomal plasmid to show that the transformed plasmid localized to only one nucleus/cell and was present at roughly 10–20 copies per nucleus. Because S. vortens grows to high densities in laboratory culture, it is a feasible diplomonad from which to purify native protein complexes. Thus, we also included a TAP tag in the plasmid constructs to permit future tagging and subsequent purification

  10. 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...

  11. 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

    -DCE back diffusion from an impervious layer. Intensive sampling (>1000 measurements) was carried out, including the withdrawal of aqueous samples at closely spaced (1 cm) outlet ports, as well as the high-resolution sampling of the source zone (agarose gel) at the end of each experiment. The transient...... behavior of the plumes originated by back diffusion was investigated by sampling the outlet ports at regular intervals in the experiments, each run for a total time corresponding to 15 pore volumes. The high-resolution sampling allowed us to resolve the spatial and temporal evolution of concentration...... [3], provided a good agreement with the experimental data...

  12. 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

  13. 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...

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

    OpenAIRE

    Guimarães,Rita de Cássia Avellaneda; Favaro,Simone Palma; Viana,Antonio Camilo Arguelho; Braga Neto,José Antônio; Neves,Valdir Augusto; Honer,Michael Robin

    2012-01-01

    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 electr...

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

    Science.gov (United States)

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

    2013-09-01

    This study compared the functional properties of serum protein concentrate (SPC) with whey protein concentrate (WPC) made from the same milk and with commercial WPC. The experimental SPC and WPC were produced at 34% or 80% protein from the same lot of milk. Protein contents of WPC and SPC were comparable; however, fat content was much lower in SPC compared with WPC and commercial WPC. The effect of drying methods (freeze vs. spray drying) was studied for 34% WPC and SPC. Few differences due to drying method were found in turbidity and gelation; however, drying method made a large difference in foam formation for WPC but not SPC. Between pH 3 and 7, SPC was found to have lower turbidity than WPC; however, protein solubility was similar between SPC and WPC. Foaming and gelation properties of SPC were better than those of WPC. Differences in functional properties may be explained by differences in composition and extent of denaturation or aggregation. Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  16. 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...

  17. 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.

  18. Short-Term Protein Stable Isotope Probing of Microbial Communities to Associate Functions with Taxa (Invited)

    Science.gov (United States)

    Lipton, M. S.; Slysz, G. W.; Steinke, L. A.; Ward, D. M.; Klatt, C. G.; Clauss, T. R.; Purvine, S. O.; Anderson, G. A.; Payne, S. H.; Bryant, D. A.

    2013-12-01

    Determining which taxa in a community perform which functions is essential for understanding metabolite fluxes and metabolic interactions among community members. Specific taxa will alter their metabolism in order to acclimate to changing environmental factors such as light through the diel cycle, changing temperature and other factors. Monitoring which proteins are being expressed, and the quantitative protein expression patterns in the individual taxa as a response to external stimuli is key to understanding these mechanisms. Protein stable isotope probing (Pro-SIP) has strong potential for revealing key metabolizing taxa in complex microbial communities. In Pro-SIP studies, label incorporation is determined by the extent of the change in the isotopic profile of peptides when measured by mass spectrometry. While most Pro-SIP work to date has been performed under controlled laboratory conditions to allow extensive isotope labeling of the target organism(s), these techniques have not been applied to short term in situ studies due to the small degree of partial labeling of the proteins. We have applied Pro-SIP to study the assimilation of a labeled substrate into proteins to determine which taxa are responsible for sequestration of dissolved inorganic carbon in microbial mats associated with the alkaline siliceous hot springs of Yellowstone National Park. This community is fueled by sunlight as it transitions from dark to light; the aim was to understand the light-dependent pathway of inorganic carbon incorporation into different taxa during the early morning hours when the mat was in low light and anoxic. Each mat sample was incubated with 13C-bicarbonate for 3 h. Substrate assimilation was determined through standard proteomic techniques along with the use of SIPPER, a collection of algorithms that sensitively measure small changes in peptide isotopic patterns, allowing the determination of which taxa assimilated the substrate during this period. For the

  19. Concord grape pomace polyphenols complexed to soy protein isolate are stable and hypoglycemic in diabetic mice.

    Science.gov (United States)

    Roopchand, Diana E; Kuhn, Peter; Krueger, Christian G; Moskal, Kristin; Lila, Mary Ann; Raskin, Ilya

    2013-11-27

    Polyphenols extracted from Concord grape pomace were stabilized by complexation to soy protein isolate (SPI) to produce grape polyphenol-SPI complex (GP-SPI) containing 5% or 10% grape polyphenols. LC-MS and MALDI-TOF analysis showed that a broad range of phytochemicals were present in the grape pomace extract. Anthocyanins and total polyphenols in the GP-SPI complex were stable after a 16-week incubation at 37 °C but were reduced by up 60% in dried grape pomace extract. Compared to vehicle (236 ± 34 mg/dL), a single dose of 300 mg/kg GP-SPI (184 ± 32 mg/dL) or 500 mg/kg GP-SPI (177 ± 28 mg/dL) having 5% grape polyphenols significantly lowered blood glucose in obese and hyperglycemic C57BL/6 mice 6 h after administration. GP-SPI allows the capture of grape pomace polyphenols in a protein-rich food matrix and may be useful as a functional food ingredient for the management of blood glucose levels.

  20. Comparison of composition and sensory properties of 80% whey protein and milk serum protein concentrates.

    Science.gov (United States)

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

    2010-05-01

    Milk serum protein concentrates (SPC) are proteins found in cheese whey that are removed directly from milk. Because SPC are not exposed to the cheese-making process, enzymatic or chemical reactions that can lead to off-flavors are reduced. The objectives of this study were to identify and compare the composition, flavor, and volatile components of 80% protein SPC and whey protein concentrates (WPC). Each pair of 80% SPC and WPC was manufactured from the same lot of milk and this was replicated 3 times. At each replication, spray-dried product from each protein source was collected. Commercial 80% WPC were also collected from several manufacturers for sensory and volatile analyses. A trained sensory panel documented the sensory profiles of the rehydrated powders. Volatile components were extracted by solid-phase microextraction and solvent extraction followed by solvent-assisted flavor evaporation with gas chromatography-mass spectrometry and gas chromatography-olfactometry. Consumer acceptance testing of acidified 6% protein beverages made with 80% SPC and WPC produced in the pilot plant and with WPC from commercial sources was conducted. The SPC was lower in fat and had a higher pH than the WPC produced in the pilot plant or commercial WPC. Few sensory differences were found between the rehydrated SPC and WPC manufactured in this study, but their flavor profiles were distinct from the flavor of rehydrated commercial WPC. The pilot-plant WPC had higher concentrations of lipid oxidation products compared with SPC, which may be related to the higher fat content of WPC. There was a large difference in appearance between 80% SPC and WPC: solutions of SPC were clear and those of WPC were opaque. Concentrations of lipid oxidation products in commercial WPC were generally higher than those in pilot-plant SPC or WPC. Sensory profiles of the peach-flavored protein beverage included cereal, free fatty acid, and soapy flavors and bitter taste in beverages made from pilot

  1. 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

  2. The effect of concentrate in meal- or pellet form on milk, protein and ...

    African Journals Online (AJOL)

    sources, on milk, protein and butterfat production of. Friesian cows was ... consume concentrate pellets much quicker than concentrate meal. The main ... Total protein kg milk produced. Days. Form of production per production per production per per concentrate. Phase. Group measured concentrate cow (kg) cow (kg).

  3. Use of human protein C concentrates in the treatment of patients with severe congenital protein C deficiency

    Directory of Open Access Journals (Sweden)

    Sabine Kroiss

    2010-03-01

    Full Text Available Sabine Kroiss, Manuela AlbisettiDivision of Hematology, University Children’s Hospital, Zurich, SwitzerlandAbstract: Protein C is one of the major inhibitors of the coagulation system that downregulate thrombin generation. Severe congenital protein C deficiency leads to a hypercoagulability state that usually presents at birth with purpura fulminans and/or severe venous and arterial thrombosis. Recurrent thrombotic events are commonly seen. From the 1990’s, several virus-inactivated human protein C concentrates have been developed. These concentrates currently constitute the therapy of choice for the treatment and prevention of clinical manifestations of severe congenital protein C deficiency. This review summarizes the available information on the use of human protein C concentrates in patients with severe congenital protein C deficiency.Keywords: Congenital protein C deficiency, protein C concentrate, purpura fulminans

  4. 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.

  5. 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.

  6. 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.

  7. Changes in body fat percentage during body weight stable conditions of increased daily protein intake vs. control.

    Science.gov (United States)

    Soenen, Stijn; Westerterp-Plantenga, Margriet S

    2010-12-02

    The objective of this study was to examine if increased protein intake vs. control influences body fat percentage during stable body weight. Body composition was assessed before and after a 3-month isoenergetic dietary intervention of 2MJ/d supplements exchanged with 2MJ/d of habitual ad libitum energy intake. The parallel design consisted of protein-rich supplements in the protein group (n=12) and an isoenergetic combination of carbohydrate and fat supplements in the control group (n=12). Daily protein intake was calculated from a 24h urinary nitrogen. Body composition was measured by a combination of underwater-weighing technique, deuterium-dilution technique and whole-body dual-energy X-ray absorptiometry (DXA), a method that allows for estimation of 4-body compartments (fat and lean; water, bone and rest). Subjects were weight stable and did not change their habitual physical activity. Daily protein intake increased in the protein group during the intervention compared to baseline with +11±14g (Pbody fat percentage showed a significant group×time interaction of decreased body fat percentage of -1.0±1.1% of the protein group vs. 0.1±0.6% of the control group (Pbody fat percentage decreased with unchanged physical activity during 3months of stable body weight. Copyright © 2010 Elsevier Inc. All rights reserved.

  8. 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

  9. 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....... Methane (CH4) production was measured the last two days in each period in open-circuit respiration chambers. Results were analysed using a linear random regression model with DM concentration as fixed effect, cow and cut number x farmer as random intercepts and with a cut number x farmer random slope...... can be improved by pre-wilting to a higher DM concentration before ensiling. Abbreviations AA, amino acids; AA-N, amino acid nitrogen; ADL, acid detergent lignin; aNDFom, neutral detergent fibre assayed with heat stable amylase and expressed exclusive of residual ash; CP, crude protein; CH4, methane...

  10. An Arg-rich putative prebiotic protein is as stable as its Lys-rich variant.

    Science.gov (United States)

    Diez-García, Fernando; Chakrabartty, Avijit; González, Carlos; Laurents, Douglas V

    2012-12-15

    An Arg-rich peptide called RIA7; sequence ac-ARAAAAAIRAIAAIIRAGGY-am, tetramerizes to form a well folded, four helix X-bundle protein. The Arg side chains are solvent exposed and the hydrophobic core is composed of the side chains from some Alas, all the Iles and the C-terminal Tyr. Since Gly, Ala and Ile, and in lesser amounts Arg and Tyr have been reported to form under putative prebiotic Earth conditions, it is plausible that RIA7-like peptides might have formed on the primitive Earth and interacted with RNAs. The interaction of RIA7 with two RNAs was tested and the formation of insoluble aggregates was observed. These results contrast with previous studies of a Lys-rich variant, called KIA7, which promotes the cleavage of RNAs. Their close structural similarity makes RIA7 and KIA7 an excellent system to compare the relative contributions of Arg and Lys to protein conformational stability. NMR-monitored hydrogen/deuterium exchange measurements and CD-monitored thermal denaturation experiments performed at different peptide and salt concentrations reveal that the conformational stabilities of RIA7 and KIA7 are practically the same. This finding has relevance for protein engineering as Lys is frequently replaced by Arg to improve ligand binding and membrane association and penetration. Copyright © 2012 Elsevier Inc. All rights reserved.

  11. 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...

  12. Effects of Energy intake and, dietary protein concentration on energy ...

    African Journals Online (AJOL)

    Increased protein intake resulted into incresed rate of heat production and protein energy retention and low rate of fat energy retention. The energy requirement for maintenance was slightly lower in animals fed on the high compared with those on low-protein diets. The partial efficiency of ME utilisation for growth, was poor ...

  13. Enhanced protein internalization and efficient endosomal escape using polyampholyte-modified liposomes and freeze concentration

    OpenAIRE

    Ahmed, Sana; Fujita, Satoshi; Matsumura, Kazuaki

    2016-01-01

    Here we show a new strategy for efficient freeze concentration-mediated cytoplasmic delivery of proteins, obtained via the endosomal escape property of polyampholyte-modified liposomes. The freeze concentration method successfully induces the efficient internalization of proteins simply by freezing cells with protein and nanocarrier complexes. However, the mechanism of protein internalization remains unclear. Here, we designed a novel protein delivery carrier by modifying liposomes through in...

  14. 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.

  15. Newcastle disease virus induces stable formation ofbona fidestress granules to facilitate viral replication through manipulating host protein translation.

    Science.gov (United States)

    Sun, Yingjie; Dong, Luna; Yu, Shengqing; Wang, Xiaoxu; Zheng, Hang; Zhang, Pin; Meng, Chunchun; Zhan, Yuan; Tan, Lei; Song, Cuiping; Qiu, Xusheng; Wang, Guijun; Liao, Ying; Ding, Chan

    2017-04-01

    Mammalian cells respond to various environmental stressors to form stress granules (SGs) by arresting cytoplasmic mRNA, protein translation element, and RNA binding proteins. Virus-induced SGs function in different ways, depending on the species of virus; however, the mechanism of SG regulation of virus replication is not well understood. In this study, Newcastle disease virus (NDV) triggered stable formation of bona fide SGs on HeLa cells through activating the protein kinase R (PKR)/eIF2α pathway. NDV-induced SGs contained classic SG markers T-cell internal antigen (TIA)-1, Ras GTPase-activating protein-binding protein (G3BP)-1, eukaryotic initiation factors, and small ribosomal subunit, which could be disassembled in the presence of cycloheximide. Treatment with nocodazole, a microtubule disruption drug, led to the formation of relatively small and circular granules, indicating that NDV infection induces canonical SGs. Furthermore, the role of SGs on NDV replication was investigated by knockdown of TIA-1 and TIA-1-related (TIAR) protein, the 2 critical components involved in SG formation from the HeLa cells, followed by NDV infection. Results showed that depletion of TIA-1 or TIAR inhibited viral protein synthesis, reduced extracellular virus yields, but increased global protein translation. FISH revealed that NDV-induced SGs contained predominantly cellular mRNA rather than viral mRNA. Deletion of TIA-1 or TIAR reduced NP mRNA levels in polysomes. These results demonstrate that NDV triggers stable formation of bona fide SGs, which benefit viral protein translation and virus replication by arresting cellular mRNA.-Sun, Y., Dong, L., Yu, S., Wang, X., Zheng, H., Zhang, P., Meng, C., Zhan, Y., Tan, L., Song, C., Qiu, X., Wang, G., Liao, Y., Ding, C. Newcastle disease virus induces stable formation of bona fide stress granules to facilitate viral replication through manipulating host protein translation. © FASEB.

  16. 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.

  17. Concentration-dependent protein adsorption at the nano-bio interfaces of polymeric nanoparticles and serum proteins.

    Science.gov (United States)

    Zhang, Tian-Xu; Zhu, Guan-Yin; Lu, Bo-Yao; Zhang, Chao-Liang; Peng, Qiang

    2017-11-01

    A comprehensive understanding of nanoparticle (NP)-protein interaction (protein corona formation) is required. So far, many factors influencing this interaction have been investigated, like size and ζ potential. However, NPs exposure concentration has always been ignored. Herein, we aim to disclose the correlation of NPs exposure concentration with protein adsorption. Four polymeric NPs systems possessing similar sizes (230 ± 20 nm) but varied ζ potentials (-30 ∼ +40 mv) were prepared. Physicochemical properties and protein adsorption upon NP-protein interaction were characterized. Protein adsorption capacity and adsorbed protein types were NPs concentration-dependent. Considering the critical impacts of protein adsorption on NPs delivery, our work could be an urgent warning about the possible risks of dosage adjustment of nanoformulations.

  18. Reducing the stiffness of concentrated whey protein isolate (WPI) gels by using WPI microparticles

    NARCIS (Netherlands)

    Purwanti, N.; Moerkens, A.; Goot, van der A.J.; Boom, R.M.

    2012-01-01

    Concentrated protein gels were prepared using native whey protein isolate (WPI) and WPI based microparticles. WPI microparticles were produced by making gel pieces from a concentrated WPI suspension (40% w/w), which were dried and milled. The protein within the microparticles was denatured and the

  19. 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.

  20. 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

  1. 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 perspectives.

  2. 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

  3. influence of dietary energy concentration on protein deposition of ad ...

    African Journals Online (AJOL)

    diet 2 (15,57 MJ ME/kg), however, consumed so much more dry matter that they actually had a higher energy intake than pigs on Treatment l. The fact that the pigs compensated for energy intake and that dietary protein content was almost constant (23D resulted in higher protein intakes by the pigs fed diets containing ...

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

    African Journals Online (AJOL)

    STORAGESEVER

    2009-07-20

    Jul 20, 2009 ... increase their applications in foods, which might offer food producers more choice in production. One approach to improve the functional properties of soybean protein products is enzymatic treatment. It could be seen from literatures that limited proteolysis of soybean protein pro- ducts offered a possibility to ...

  5. 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

  6. Alfalfa Protein Concentrate as a Less Ruminally Degradable Protein Source for High-Producing Dairy Cows

    Directory of Open Access Journals (Sweden)

    C.D. Lu

    1997-01-01

    Full Text Available Ten multiparous Holstein cows were used in a 105-day continuous feeding trial to compare soybean meal (SBM and alfalfa protein concentrate (APC as supplemental protein sources for high-producing dairy cows. Dairy cows with an average milk production of 35.2 kg/day were paired and randomly assigned to one of the treatments. A double reversal design was employed With 35 days per period. The comparison between treatments was made during the last 21 days of each period. Experimental diets, containing 60% corn silage and 10% alfalfa hay, were formulated to be isonitrogenous and isoenergetic (crude protein. 15.6%; net energy tor lactation, 1.60 Mcal/kg DM. Fifty percent of dietary nitrogen was provided by test proteins. Animals were fed four times daily. Chromie oxide boluses, 50 g/day were used to estimate digestion coefficients. Ruminal protein degradability in viva (% and in situ(%were 56.8, 68.3 and 47.3. 35.3 for SBM and APC, respectively. Dry matter intake (kg/day was 19.1 and 18.6 for cows fed SBM and APC. respectively. Rummal parameters cows fed SBM vs APC were, pH. 6.6 vs 6.7, ammonia nitrogen(mg/dI 10.8 vs 6.2(p less than 0.01; total volatile fatty acids (mM, 86.3 VS 84.6(P LESS THAN 0.05, acetate (% , 50.8 vs 64.9 (p less than 0.01; propionate dry matter , 65.3 vs 61.8; crude protein . 67.6 vs 61.1 (P less than 0.05 neutral detergent fiber , 51.1 vs 52.5 ; acid detergent fiber, 47.7 vs 49.0 . A slightly higher (P more than 0.05 amount of milk was produced by cows fed APC(28.6+_ 3.5 vs 28.2+- 3.8 kg/day. Each additional gramof undergradable intake protein provided by APC continued to a 5- gram increase in 4% fat-corrected milk production.

  7. 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

  8. The nucleotide-free state of heterotrimeric G proteins α-subunit adopts a highly stable conformation.

    Science.gov (United States)

    Andhirka, Sai Krishna; Vignesh, Ravichandran; Aradhyam, Gopala Krishna

    2017-08-01

    Deciphering the mechanism of activation of heterotrimeric G proteins by their cognate receptors continues to be an intriguing area of research. The recently solved crystal structure of the ternary complex captured the receptor-bound α-subunit in an open conformation, without bound nucleotide has improved our understanding of the activation process. Despite these advancements, the mechanism by which the receptor causes GDP release from the α-subunit remains elusive. To elucidate the mechanism of activation, we studied guanine nucleotide-induced structural stability of the α-subunit (in response to thermal/chaotrope-mediated stress). Inherent stabilities of the inactive (GDP-bound) and active (GTP-bound) forms contribute antagonistically to the difference in conformational stability whereas the GDP-bound protein is able to switch to a stable intermediate state, GTP-bound protein loses this ability. Partial perturbation of the protein fold reveals the underlying influence of the bound nucleotide providing an insight into the mechanism of activation. An extra stable, pretransition intermediate, 'empty pocket' state (conformationally active-state like) in the unfolding pathway of GDP-bound protein mimics a gating system - the activation process having to overcome this stable intermediate state. We demonstrate that a relatively more complex conformational fold of the GDP-bound protein is at the core of the gating system. We report capturing this threshold, 'metastable empty pocket' conformation (the gate) of α-subunit of G protein and hypothesize that the receptor activates the G protein by enabling it to achieve this structure through mild structural perturbation. © 2017 Federation of European Biochemical Societies.

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

    Science.gov (United States)

    Serret, María D.; Yousfi, Salima; Vicente, Rubén; Piñero, María C.; Otálora-Alcón, Ginés; del Amor, Francisco M.; Araus, José L.

    2018-01-01

    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 relationship with N

  10. 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

  11. Bioaccumulation of perfluoroalkyl substances by Daphnia magna in water with different types and concentrations of protein.

    Science.gov (United States)

    Xia, Xinghui; Rabearisoa, Andry H; Jiang, Xiaoman; Dai, Zhineng

    2013-10-01

    Perfluoroalkyl substances (PFASs) are sometimes regarded as proteinophilic compounds, however, there is no research report about the effect of environmental protein on the bioaccumulation of PFASs in waters. In the present study we investigated influences of protein on the bioaccumulation of six kinds of PFASs by Daphnia magna in water; it included perfluorooctane sulfonate, perfluorooctanoic acid, perfluorononanoic acid, perfluorodecanoic acid, perfluoroundecanoic acid, and perfluorododecanoic acid. Two types of protein including bovine albumin from animal and soy peptone from plant were compared and the effects of protein concentration were investigated. Both types of protein at high concentrations (10 and 20 mg L(-1)) suppressed the bioaccumulation of PFASs. When protein concentration increased from 0 to 20 mg L(-1), the decreasing ratios of the PFAS body burden (35.3-52.9%) in Daphnia magna induced by bovine albumin were significantly higher than those (22.0-36.6%) by soy peptone. The dialysis bag experiment results showed that the binding of PFASs to protein followed the Freundlich isotherm, suggesting it is not a linear partitioning process but an adsorption-like process. The partition coefficients of PFASs between bovine albumin and water were higher compared to soy peptone; this resulted in higher reducing rates of freely dissolved concentrations of PFASs with increasing bovine albumin concentration, leading to a stronger suppression of PFAS bioaccumulation. However, the presence of both types of protein with a low concentration (1 mg L(-1)) enhanced the bioaccumulation of PFASs. Furthermore, the water-based bioaccumulation factor based on the freely dissolved concentrations of PFASs even increased with and the depuration rate constants of PFASs from Daphnia magna decreased with protein concentration, suggesting that protein would not only reduce the bioavailable concentrations and uptake rates of PFASs but also lower the elimination rates of PFASs in

  12. Consumer Acceptance of Seaweed Ball (Eucheuma Cottonii) Fortified with Catfish Protein Concentrate (Pangasius Hypophthalmus)

    OpenAIRE

    Naibaho, Marianche F; ', Dewita; ', Edison

    2014-01-01

    The study was conducted to evaluate consumer acceptance of seaweed ball fortified with catfish protein concentrate . About 5 kg seaweed was taken from a fish market in Pekanbaru. Three types of seaweed balls were prepared from seaweed flour which was fortificated respectively with 5%, 10%, and 15% catfish protein concentrate. The seaweed balls were evaluated for consumer acceptance and chemical composition. The results indicate that the seaweed balls added with 15% catfish protein concentrate...

  13. 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.

  14. Proteomic analysis of differential proteins in pancreatic carcinomas: Effects of MBD1 knock-down by stable RNA interference

    Directory of Open Access Journals (Sweden)

    Ni Quanxing

    2008-04-01

    Full Text Available Abstract Background Methyl-CpG binding domain protein 1 (MBD1, a suppressor of gene transcription, may be involved in inactivation of tumor suppressor genes during tumorigenesis. Over-expression of MBD1 has been reported in human pancreatic carcinomas. Methods In this study, we established a MBD1-knock-down pancreatic cancer cell line (BxPC-3 using stable RNA interference, to compare the proteomic changes between control and MBD1-knock-down cells using two-dimensional gel electrophoresis and mass spectrometry. Results We identified five proteins that were up-regulated and nine proteins that were down-regulated. Most of the identified proteins are involved in tumorigenesis, some are prognostic biomarkers for human malignant tumors. Conclusion Our data suggest that these differential proteins may be associated with the function of MBD1, and provide some insight into the functional mechanism of MBD1 in the development of pancreatic cancer.

  15. The effect of different rumen degradable protein concentrations and ...

    African Journals Online (AJOL)

    Nolte Joubert

    indicated that increasing RDP levels tended to decrease arginine, but significantly increased tryptophan concentrations. Histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, valine and total essential AA concentrations were not affected by dietary RDP level. Increasing the urea-N content of RDP.

  16. 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.

  17. 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.

  18. Improved protein synthesis and secretion through medium enrichment in a stable recombinant yeast strain.

    Science.gov (United States)

    Wang, Z; Da Silva, N A

    1993-06-05

    Two Saccharomyces cerevisiae strains were employed to investigate the effects of medium enrichment on the expression and secretion of a recombinant protein. One was a stable autoselection strain with mutations in the ura3, fur1, and urid-k genes. The combination of these three mutations blocks both the pyrimidine nucleotide biosynthetic and salvage pathways and is lethal to the cells. Retention of the plasmid, which carries a URA3 gene, was essential for cell viability. Therefore, all media were selective, allowing cultivation of the strain in complex medium. The second strain was a nonautoselection (control) strain and is isogenic to the first except for the fur1 and urid-k mutations. The plasmid utilized contains the yeast invertase gene under the control of the MFalpha1 promoter and leader sequence. The expression and secretion of invertase for the autoselection strain were examined in batch culture for three media: a minimal medium (SD), a semidefined medium (SDC), and a rich complex medium (YPD). Biomass yields and invertase productivity (volumetric activity) increased with the complexity of the medium; total invertase volumetric activity in YPD was 100% higher than in SDC and 180% higher than in SD. Specific activity, however, was lowest in the SDC medium. Secretion efficiency was extremely high in all three media; for the majority of the culture, 80-90% of the invertase was secreted into the periplasmic space and/or culture medium. A glucose pulse at the end of batch culture in YPD facilitated the transport of residual cytoplasmic invertase. For the nonautoselection strain, invertase productivity did not improve as the medium was enriched from SDC to YPD, and plasmid stability in the complex YPD medium dropped from 54% to 34% during one batch fermentation. During long-term sequential batch culture in YPD, invertase activity decreased by 90% and the plasmid-containing fraction dropped from 56% to 8.8% over 44 generations of growth. The expression level for

  19. 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.

  20. The relative nutritive value of lucerne leaf protein concentrate (LPC ...

    African Journals Online (AJOL)

    The hot-water method dilutes these water- soluble substances and could, therefore, yield a LPC with a better protein quality thari the steam-treatment method. Furthermore, the temperature of the steam at the point of injection may be high enough ...

  1. 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 ...

  2. Concentration and desalination of protein solutions by ultrafiltration

    NARCIS (Netherlands)

    Noordman, T.R; Ketelaar, T.H.; Donkers, F.; Wesselingh, J.A

    In this study the effects of ionic strength and PH on ultrafiltration of protein solutions are examined. Experiments are done with BSA/NaCl solutions with PH varying from 3 to 8 and ionic strength from 5 to 150 mM. Fluxes depend on solution conditions and increase with decreasing ionic strength.

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

    African Journals Online (AJOL)

    Jane

    2011-10-05

    Oct 5, 2011 ... 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.

  4. Plasma protein concentrations of the young and adult Amazona brasiliensis parrots

    Directory of Open Access Journals (Sweden)

    Schmidt Elizabeth Moreira Dos Santos

    2017-01-01

    Full Text Available Introduction. The Red-tailed Amazon parrot (Amazona brasiliensis is an endangered species of the Psittacine family, and for which various data are important for a comprehensive preservation plan. Data about plasma protein gel electrophoresis of Amazon parrot blood are scarce. The purpose of this study was to determine plasma protein concentrations and concentrations of major protein bands in blood of young and adult Red-tailed Amazon parrot (Amazona brasiliensis. Materials and Methods. Blood samples from eight young and eight adult healthy free-living parrots were obtained. Plasma protein concentration and fractions were determined using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE. Mann-Whitney U test was used to compare variables. Results and Conclusions. Six major protein bands with the following molecular weights were identified by SDS-PAGE: 170 kDa, 117 kDa, 85 kDa (putative ovotransferrin, 60 kDa, 45 kDa and 23 kDa. Adult parrots had significantly higher concentrations of total proteins, albumin and other proteins with similar mobility (around 60 kDa. Young birds had significantly higher levels of 23kDa proteins. The concentration of putative ovotransferrin (85 kDa was not different between young and adult parrots. Plasma protein gel electrophoresis patterns in Red-tailed Amazon parrots are similar between young and adult animals, but specific protein bands differ in their absolute concentrations. This finding should be taken into consideration when clinical pathology data are analysed.

  5. Cortical concentrations of metals and protein in the brain of two ...

    African Journals Online (AJOL)

    The male RS goats had significantly higher levels (P<0.05) of Mn than the females. While the male WAD goats had higher levels of total crude protein, the concentration between the sexes of the RS goats was the same. This study has given information on metal and protein concentration in the brain of goat for use in ...

  6. 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.

  7. Chemical composition, functional and pasting properties of cassava starch and soy protein concentrate blends

    OpenAIRE

    Chinma, Chiemela Enyinnaya; Ariahu, Charles Chukwuma; Abu, Joseph Oneh

    2011-01-01

    The chemical, functional and pasting properties of cassava starch and soy protein concentrate blends intended for biofilm processing were studied. Cassava starch and soy protein concentrates were prepared and mixed at different proportions (100: 0%; 90 : 10%; 80 : 20%; 70 : 30%; 60;40% and 50: 50%). Addition of varying levels of soy protein concentrates to cassava starch led to increases in moisture (from 7.10 to 9.17%), protein ( from 0.32 to 79.03%), ash (from 0.45 to 2.67%) and fat (from 0...

  8. Heat shock protein concentration and clarity of porcine lenses incubated at elevated temperatures.

    Science.gov (United States)

    Dzialoszynski, T M; Milne, K J; Trevithick, J R; Noble, E G

    2016-01-01

    To quantify the concentration of heat shock proteins in lenses in lens organ culture at elevated temperatures, and to examine the relation between elevated temperature and lens clarity. Pig lenses obtained from a local abattoir were dissected aseptically and incubated in medium M199 without serum for 4 days to stabilize, and lenses with protein leakage of less than 10 mg/l were obtained for heat shock exposure. Heat shock was performed by incubation for 1 h in M199 without serum at various temperatures ranging from 37 °C to 55 °C. After incubation for 24 h, cataract blurring of the images was assessed using Scantox™ and Scion Image analysis of the lens photographs. Lens homogenates were subsequently analyzed for Hsp70 and Hsp27 with western blotting. The degree of cataract blurring of the images increased with increasing temperature, but the two functional measures provided different results. Focal length inconsistency, as assessed with the back vertex distance standard error of the mean (BVD SEM; the variability in focal lengths measured at 20 equally spaced locations across the lens, Scantox™), increased nearly linearly with the heat treatment temperature. In contrast, decreased clarity, evident by a fuzzy image with lower contrast, was not markedly altered as the temperature rose until a threshold of approximately 47.5 °C. The inducible isoform of the Hsp70 family (Hsp70) of heat shock proteins was increased at all temperatures above the control except those above 50 °C. Changes in Hsp27 were less clear as the protein content increased only at the incubation temperatures of 39 °C and 48.5 °C. The porcine lens demonstrates subtle changes in the variability of the focal length, and the variability increases as the incubation temperature rises. In contrast, lens clarity is relatively stable at temperatures up to 47.5 °C, above which dramatic changes, indicative of the formation of cataracts, occur. The lens content of Hsp70 was elevated in lenses

  9. 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...

  10. 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

  11. 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.

  12. Stable expression of a GFP-BSD fusion protein in Babesia bovis merozoites

    Science.gov (United States)

    Transfection has been a valuable technique for elucidating gene function in many pathogens. While transient transfection of Babesia spp has been reported previously, stable integration of exogenous genes in Babesia has proven difficult. In this study, a plasmid was designed to target integration of ...

  13. 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...

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

    OpenAIRE

    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 indic...

  15. Soybean bio-refinery platform: enzymatic process for production of soy protein concentrate, soy protein isolate and fermentable sugar syrup.

    Science.gov (United States)

    Loman, Abdullah Al; Islam, S M Mahfuzul; Li, Qian; Ju, Lu-Kwang

    2016-10-01

    Soybean carbohydrate is often found to limit the use of protein in soy flour as food and animal feed due to its indigestibility to monogastric animal. In the current study, an enzymatic process was developed to produce not only soy protein concentrate and soy protein isolate without indigestible carbohydrate but also soluble reducing sugar as potential fermentation feedstock. For increasing protein content in the product and maximizing protein recovery, the process was optimized to include the following steps: hydrolysis of soy flour using an Aspergillus niger enzyme system; separation of the solid and liquid by centrifugation (10 min at 7500×g); an optional step of washing to remove entrapped hydrolysate from the protein-rich wet solid stream by ethanol (at an ethanol-to-wet-solid ratio (v/w) of 10, resulting in a liquid phase of approximately 60 % ethanol); and a final precipitation of residual protein from the sugar-rich liquid stream by heat treatment (30 min at 95 °C). Starting from 100 g soy flour, this process would produce approximately 54 g soy protein concentrate with 70 % protein (or, including the optional solid wash, 43 g with 80 % protein), 9 g soy protein isolate with 89 % protein, and 280 ml syrup of 60 g/l reducing sugar. The amino acid composition of the soy protein concentrate produced was comparable to that of the starting soy flour. Enzymes produced by three fungal species, A. niger, Trichoderma reesei, and Aspergillus aculeatus, were also evaluated for effectiveness to use in this process.

  16. 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) tryptophanate...... [EMIM][TRP] (5 mol% in water). Upon analyzing the radius of gyration, the solvent-accessible surface area, root-mean-squared deviations, and inter- and intramolecular hydrogen bonds, we found that the mini-protein remains stable at 30–40 K higher temperatures in aqueous amino acid based ionic liquids...... (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.]...

  17. Protein freeze concentration and micro-segregation analysed in a temperature-controlled freeze container

    Directory of Open Access Journals (Sweden)

    Ulrich Roessl

    2015-06-01

    Full Text Available To examine effects of varied freezing conditions on the development of spatial heterogeneity in the frozen protein solution, macroscopic freeze concentration and micro-segregation of bovine serum albumin (BSA were investigated in a temperature-controlled 200-ml freeze container. Freezing to −40 °C promoted formation of protein concentration gradients (69–114 μg ml−1 in frozen samples taken from 12 different freezer positions, whereby slow freezing in 4 h or longer facilitated the evolution of strong spatial heterogeneities and caused local concentration increases by 1.15-fold relative to the initial protein concentration (100 μg ml−1. To visualize protein micro-segregation during phase separation, BSA was conjugated with fluorescein isothiocyanate and confocal laser scanning fluorescence microscopy was used to localize and size the freeze-concentrated protein regions. Slow freezing resulted in distinctly fewer and larger protein domains in the frozen bulk than fast freezing. Surface stress on the protein during freezing would therefore be minimized at low cooling rates; microscopic freeze concentration would however be highest under these conditions, potentially favoring protein aggregation.

  18. 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.

  19. Composition, Structure and Functional Properties of Protein Concentrates and Isolates Produced from Walnut (Juglans regia L.

    Directory of Open Access Journals (Sweden)

    Xiaoying Mao

    2012-02-01

    Full Text Available 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. H0 of WPC was significantly higher ( p < 0.05 than those of DFWF and WPI, whilst WPI had a higher H0 compared to DFWF. The secondary 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.

  20. 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 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.

  1. Using magnetic nanoparticles to probe protein damage in ferritin caused by freeze concentration

    Directory of Open Access Journals (Sweden)

    E. F. Chagas

    2015-11-01

    Full Text Available We demonstrate a method for monitoring the damage caused to a protein during freeze-thawing in the presence of glycerol, a cryo-protectant. For this work we synthesized magnetite nanoparticles doped with 2.5% cobalt inside the protein ferritin (CMF, dissolved them in different concentration glycerol solutions and measured their magnetization after freezing in a high applied field (5 T. As the temperature was raised, a step-like decrease in the sample magnetization was observed, corresponding to the onset of Brownian relaxation as the viscosity of the freeze-concentrated glycerol solution decreased. The position of the step reveals changes to the protein hydrodynamic radius that we attribute to protein unfolding, while its height depends on how much protein is trapped by ice during freeze concentration. Changes to the protein hydrodynamic radius are confirmed by dynamic light scattering (DLS measurements, but unlike DLS, the magnetic measurements can provide hydrodynamic data while the solution remains mainly frozen.

  2. 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...

  3. 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.

  4. 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.

  5. Validation of a single biopsy approach and bolus protein feeding to determine myofibrillar protein synthesis in stable isotope tracer studies in humans

    Directory of Open Access Journals (Sweden)

    Baker Steven K

    2011-03-01

    Full Text Available Abstract Background Minimizing the number of muscle biopsies has important methodological implications and minimizes subject discomfort during a stable isotope amino acid infusion. We aimed to determine the reliability of obtaining a single muscle biopsy for the calculation of muscle protein fractional synthetic rate (FSR as well as the amount of incorporation time necessary to obtain that biopsy after initiating a stable isotope infusion (Study 1. The calculation of muscle protein FSR requires tracer steady-state during the stable isotope infusion. Therefore, a second aim was to examine if steady-state conditions are compromised in the precursor pools (plasma free or muscle intracellular [IC] after ingestion of a tracer enriched protein drink and after resistance exercise (Study 2. Methods Sixteen men (23 ± 3 years; BMI = 23.8 ± 2.2 kg/m2, means ± SD were randomized to perform Study 1 or Study 2 (n = 8, per study. Subjects received a primed, constant infusion of L-[ring-13C6]phenylalanine coupled with muscle biopsies of the vastus lateralis to measure rates of myofibrillar protein synthesis (MPS. Subjects in Study 2 were fed 25 g of whey protein immediately after an acute bout of unilateral resistance exercise. Results There was no difference (P = 0.3 in rates of MPS determined using the steady-state precursor-product equation and determination of tracer incorporation between sequential biopsies 150 min apart or using plasma protein as the baseline enrichment, provided the infusion length was sufficient (230 ± 0.3 min. We also found that adding a modest amount of tracer (4% enriched, calculated based on the measured phenylalanine content of the protein (3.5% in the drink, did not compromise steady-state conditions (slope of the enrichment curve not different from zero in the plasma free or, more importantly, the IC pool (both P > 0.05. Conclusions These data demonstrate that the single biopsy approach yields comparable rates of muscle

  6. Validation of a single biopsy approach and bolus protein feeding to determine myofibrillar protein synthesis in stable isotope tracer studies in humans.

    Science.gov (United States)

    Burd, Nicholas A; West, Daniel Wd; Rerecich, Tracy; Prior, Todd; Baker, Steven K; Phillips, Stuart M

    2011-03-09

    Minimizing the number of muscle biopsies has important methodological implications and minimizes subject discomfort during a stable isotope amino acid infusion. We aimed to determine the reliability of obtaining a single muscle biopsy for the calculation of muscle protein fractional synthetic rate (FSR) as well as the amount of incorporation time necessary to obtain that biopsy after initiating a stable isotope infusion (Study 1). The calculation of muscle protein FSR requires tracer steady-state during the stable isotope infusion. Therefore, a second aim was to examine if steady-state conditions are compromised in the precursor pools (plasma free or muscle intracellular [IC]) after ingestion of a tracer enriched protein drink and after resistance exercise (Study 2). Sixteen men (23 ± 3 years; BMI = 23.8 ± 2.2 kg/m2, means ± SD) were randomized to perform Study 1 or Study 2 (n = 8, per study). Subjects received a primed, constant infusion of L-[ring-13C6]phenylalanine coupled with muscle biopsies of the vastus lateralis to measure rates of myofibrillar protein synthesis (MPS). Subjects in Study 2 were fed 25 g of whey protein immediately after an acute bout of unilateral resistance exercise. There was no difference (P = 0.3) in rates of MPS determined using the steady-state precursor-product equation and determination of tracer incorporation between sequential biopsies 150 min apart or using plasma protein as the baseline enrichment, provided the infusion length was sufficient (230 ± 0.3 min). We also found that adding a modest amount of tracer (4% enriched), calculated based on the measured phenylalanine content of the protein (3.5%) in the drink, did not compromise steady-state conditions (slope of the enrichment curve not different from zero) in the plasma free or, more importantly, the IC pool (both P > 0.05). These data demonstrate that the single biopsy approach yields comparable rates of muscle protein synthesis, provided a longer incorporation time is

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

    Science.gov (United States)

    Moghadam, Ali; Niazi, Ali; Afsharifar, Alireza; Taghavi, Seyed Mohsen

    2016-01-01

    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. PMID:27459300

  8. 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.

  9. Practice on improving fattening local cattle production in Vietnam by increasing crude protein level in concentrate and concentrate level.

    Science.gov (United States)

    Van Dung, Dinh; Ba, Nguyen Xuan; Van, Nguyen Huu; Phung, Le Dinh; Ngoan, Le Duc; Cuong, Vu Chi; Yao, Wen

    2013-10-01

    Two experiments were conducted to determine the effects of crude protein (CP) level in concentrate (experiment 1) and concentrate level (experiment 2) on feed intake, nutrient digestibility, nitrogen (N) retention, ruminal pH and NH3-N concentration and average daily gain (ADG) of Vietnamese local fattening cattle. Animals (24 cattle, initial live weight (LW) 150.3 ± 11.8 kg in experiment 1 and 145.1 ± 9.8 kg in experiment 2) were allotted based on LW to one of four treatments in a randomised complete block design. In experiment 1, concentrate with four levels of CP (10, 13, 16 and 19 %) was fed at 1.5 % of LW. In experiment 2, concentrate was fed at 1.0, 1.4, 1.8 and 2.2 % of LW. In both experiments, roughage was 5 kg/day native grass and ad libitum rice straw (fresh basis). Results showed that the CP level in concentrate significantly affected dry matter (DM) intake (P matter (OM) and neutral detergent fibre (NDF) digestibility (P > 0.05), whereas CP digestibility increased (P 0.05). OM digestibility and NH3-N concentration increased linearly (P consumption (P cattle in Vietnam.

  10. 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.

  11. Compatibility of preparatory procedures for the analysis of cortisol concentrations and stable isotope (δ13C, δ15N) ratios: a test on brown bear hair

    Science.gov (United States)

    Sergiel, Agnieszka; Hobson, Keith A.; Janz, David M.; Cattet, Marc; Selva, Nuria; Kapronczai, Luciene; Gryba, Chantel

    2017-01-01

    Abstract The measurement of naturally occurring glucocorticoids and stable isotopes of several elements has gained importance in wildlife studies in recent decades and opened a myriad of ecological applications. Cortisol and stable isotopes equilibrate in animal tissues over periods of integration related to the growth rate of the tissue, providing information reflecting systemic cortisol secretion and dietary intake. Sample preparation shares the common step of first cleaning the sample of external contamination. However, it is not well understood how different solvents used in sample preparation affect isotopic and cortisol values, and whether it is safe to follow the same procedures for both measures to optimize analyses of the same sample. We conducted an experiment to compare different preparation protocols for the analysis of cortisol concentrations and stable carbon (δ13C) and nitrogen (δ15N) isotope ratios in hair. Hair samples from 12 brown bears (Ursus arctos) were each divided into five aliquots; two aliquots were rinsed with a 2:1 chloroform:methanol (v/v) mixture with one aliquot ground prior to cortisol analysis and the other left intact for stable isotope analyses; two aliquots were washed with methanol with one aliquot ground prior to cortisol analysis and the other left intact for stable isotope analyses; and one aliquot washed with methanol and ground prior to stable isotope analyses. The cortisol, δ13C and δ15N values remained consistent following all treatments. Our results indicate that hair samples rinsed with a 2:1 chloroform:methanol mixture or washed with methanol can be used for both types of analyses. Further, hair that has been ground in a standard hair cortisol procedure can also be used for stable isotope analysis. This information is important for improving laboratory efficiency and compatibility of procedures used for wildlife physiological ecology studies where concurrent measurements of cortisol and stable isotopes in hair are

  12. Grain protein concentration and harvestable protein under future climate conditions. A study of 108 spring barley accessions.

    Science.gov (United States)

    Ingvordsen, Cathrine H; Gislum, René; Jørgensen, Johannes R; Mikkelsen, Teis N; Stockmarr, Anders; Jørgensen, Rikke B

    2016-04-01

    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 concentration by 5%, while elevated temperature (+5 °C day/night) substantially increased protein concentration by 29%. The combined treatment increased protein concentration across accessions by 8%. This was an increase less than predicted from strictly additive effects of the individual treatments. Despite the increase in grain protein concentration, the decrease in grain yield at combined elevated temperature and elevated [CO2] resulted in 23% less harvestable protein. There was variation in the response of the 108 accessions, which might be exploited to at least maintain if not increase harvestable grain protein under future climate change conditions. © The Author 2016. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  13. 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.

  14. 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.

  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. Stable isotope labeling method for the investigation of protein haptenation by electrophilic skin sensitizers.

    Science.gov (United States)

    Parkinson, Erika; Boyd, Pete; Aleksic, Maja; Cubberley, Richard; O'Connor, David; Skipp, Paul

    2014-11-01

    The risk of contact sensitization is a major consideration in the development of new formulations for personal care products. However, developing a mechanistic approach for non-animal risk assessment requires further understanding of haptenation of skin proteins by sensitizing chemicals, which is the molecular initiating event causative of skin sensitization. The non-stoichiometric nature of protein haptenation results in relatively low levels of modification, often of low abundant proteins, presenting a major challenge for their assignment in complex biological matrices such as skin. Instrumental advances over the last few years have led to a considerable increase in sensitivity of mass spectrometry (MS) techniques. We have combined these advancements with a novel dual-labeling/LC-MS(E) approach to provide an in-depth direct comparison of human serum albumin (HSA), 2,4-dinitro-1-chlorobenzene (DNCB), 5-chloro-2-methyl-4-isothiazolin-3-one (MCI), trans-cinnamaldehyde, and 6-methyl coumarin. These data have revealed novel insights into the differences in protein haptenation between sensitizers with different reaction mechanisms and sensitizing potency; the extreme sensitizers DNCB and MCI were shown to modify a greater number of nucleophilic sites than the moderate sensitizer cinnamaldehyde; and the weak/non-sensitizer 6-methyl coumarin was restricted to only a single nucleophilic residue within HSA. The evaluation of this dual labeling/LC-MS(E) approach using HSA as a model protein has also demonstrated that this strategy could be applied to studying global haptenation in complex mixtures of skin-related proteins by different chemicals. © The Author 2014. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  17. Effect of treatments and washing cycles on the quality of Nile tilapia (Oreochromis niloticus protein concentrate

    Directory of Open Access Journals (Sweden)

    Lígia Boarin Alcalde

    2017-08-01

    Full Text Available The extraction of proteins from wastes reduces production costs and environmental pollution. The aim of this work was to evaluate the effect of two treatments involving decanting/sieving or centrifugation and the number of washing cycles on the quality of protein concentrate obtained from mechanically separated meat (MSM of Nile tilapia (Oreochromis niloticus. Results were analyzed in terms of final yield and proximate composition (moisture, protein, fat and ash after each washing cycle. Moisture did not vary statistically with the treatments and after the third washing cycle. However, the process involving centrifugation was more efficient for protein concentration because the final protein content increased 2.0 folds (79.82%, dry basis and fat decreased 6.1 folds (8.29%, dry basis. After four washing cycles, it was obtained a protein concentrate with 79.82% protein, 8.29% lipid and 0.45% ash (dry basis, and 80.0% yield, using the centrifugation procedure. Visual whiteness was highly improved after four washing cycles using both processes. It was concluded that the centrifugation process with four washing cycles was the most appropriate method for producing protein concentrate from MSM of Nile tilapia.

  18. Interferon gamma-induced protein-10 concentrations in children with previous tuberculosis infections and disease

    NARCIS (Netherlands)

    Bihari, Smiti; Cavalcanti, Nara; Correia, Jailson B.; Alves, Gerlane; Souza, Edvaldo; Brabin, Bernard J.; Cuevas, Luis E.

    2012-01-01

    Interferon gamma-induced protein-10 is a diagnostic test for tuberculosis infection. There is no information on its concentrations over time. Interferon gamma-induced protein-10 was evaluated in 72 children formerly treated or in former contact with tuberculosis using Quantiferon Gold in-tube. Cases

  19. 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

  20. 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

  1. Effects of enzymatic hydrolysis on the allergenicity of whey protein concentrates.

    OpenAIRE

    Cuicui Duan; Lijie Yang; Aili Li; Rui Zhao; Guicheng Huo

    2014-01-01

    Cow's milk whey consists of many protein components and some of them are antigens to human and known to modulate immune responses. Enzymatic hydrolysis is a useful method to modify proteins with allergenicity. The objective of this study was to identify whether the in vitro enzymatic hydrolysis could reduce the allergenicity of whey protein concentrates (WPC). In this study, WPC were hydrolyzed by trypsin and twenty-four BALB/c mice were divided into three groups and fed with WPC formula and ...

  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. 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

    by the expression of PML-RAR alpha. We report that PML colocalizes with the nonphosphorylated fraction of the retinoblastoma protein (pRB) within nuclear bodies and that pRB is delocalized by PML-RAR alpha expression. Both PML and PML-RAR alpha form complexes with the nonphosphorylated form of pRB in vivo...

  4. 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.

  5. 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)

  6. Chemical composition, functional and pasting properties of cassava starch and soy protein concentrate blends.

    Science.gov (United States)

    Chinma, Chiemela Enyinnaya; Ariahu, Charles Chukwuma; Abu, Joseph Oneh

    2013-12-01

    The chemical, functional and pasting properties of cassava starch and soy protein concentrate blends intended for biofilm processing were studied. Cassava starch and soy protein concentrates were prepared and mixed at different proportions (100: 0%; 90 : 10%; 80 : 20%; 70 : 30%; 60;40% and 50: 50%). Addition of varying levels of soy protein concentrates to cassava starch led to increases in moisture (from 7.10 to 9.17%), protein ( from 0.32 to 79.03%), ash (from 0.45 to 2.67%) and fat (from 0.17 to 0.98%) contents while crude fiber, carbohydrate and amylose contents decreased from ( 1.19 to 0.38%, 90.77 to 57.01% and 29.45 to 23.04%) respectively . Water absorption capacity and swelling power of cassava starch were improved as a result of soy protein concentrate addition while syneresis and solubility value of composite blends were lower than 100% cassava starch. In general, cassava-soy protein concentrate blends formed firmer gels than cassava starch alone. There were significant (p ≤ 0.05) increases in peak viscosity (from 160.12 to 268.32RVU), final viscosity (from 140.41 to 211.08RVU) and pasting temperature (from 71.00 to 72.32 °C ) of cassava starch due to addition of soy protein concentrate. These results suggest that the addition of soy protein concentrate to cassava starch affected the studied functional properties of cassava starch as evidenced by changes such as reduced syneresis, and solubility that are desirable when considering this biopolymer as an edible biofilm.

  7. 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.

  8. 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

  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

    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...... cultured for 25 generations under strong and slightly toxic expression after which only limited reduction in fluorescence was detectable. Such non-recombinogenic GFPs can help quantify intracellular responses operating a low copy number in recombination-prone organisms....

  10. 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

    ) 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...

  11. 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.

  12. 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.

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

    Science.gov (United States)

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

    2011-02-01

    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.

  15. 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.

  16. 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.

  17. 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

  18. Characterization of high-milk-protein powders upon rehydration under various salt concentrations.

    Science.gov (United States)

    Hussain, R; Gaiani, C; Aberkane, L; Scher, J

    2011-01-01

    Rehydration of native micellar casein and native whey isolate protein powders was followed in different ionic environments. Solutions of NaCl and CaCl2 in the concentration range of 0 to 12% (wt%) were used as rehydration media. The rehydration profiles obtained were interpreted in terms of wetting, swelling, and dispersion stages by using a turbidity method. Two behaviors were observed depending on the salt concentration. For native micellar casein powder, a significant change was observed between 3 and 6% NaCl and between 0.75 and 1.5% CaCl2. The first behavior (low salt concentration) presents a typical rehydration profile: quick wetting, swelling, and long dispersion stage. The dispersion stage of the second behavior (high salt concentration) was significantly shortened, indicating a strong modification of the protein backbone. The rehydration of whey protein powder was less influenced by salts. At low salt concentrations, a typical profile for whey powders was observed: wetting with lump formation and no swelling followed by a quick dispersion. At high CaCl2 concentrations, no turbidity stabilization was observed, indicating a possible protein unfolding and denaturation. Additionally, the changes in secondary structures of the 2 proteins upon salt increase were followed by Fourier transform infrared spectroscopy and confirmed the different profiles observed. Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  19. A simple, disposable microfluidic device for rapid protein concentration and purification via direct-printing.

    Science.gov (United States)

    Yu, Hui; Lu, Yu; Zhou, Yi-ge; Wang, Feng-bin; He, Feng-yun; Xia, Xing-hua

    2008-09-01

    A facile and disposable microfluidic device for rapid protein concentration was fabricated by using a direct printing process. Two printed V-shaped microchannels in mirror image orientation were separated by a 100 mum wide toner gap. When a high electric field was applied across the two channels, nanofissures were formed by electric breakdown at the junction toner gap. This microfluidic device with nanofissures was used as a concentrator for protein. Negatively charged proteins were observed to concentrate at the anode side of the nanofissures upon application of an electric field across this junction. Using this device, about 10(3)-10(5)-fold protein concentration was achieved within 10 min. Systematic investigation showed that the concentration mechanism could be explained by the ion exclusion-enrichment effect of the nanofissures. In addition, the present microchip device integrated both functions of concentration and purification were confirmed. This simple on chip protein preconcentration and purification device could be a disposable sample preparation component in printed microfluidic systems used for practical biochemical assays.

  20. 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

  1. 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.

  2. IMPACT OF DYNAMICAL HYDRATION SHELL AROUND HA PROTEIN ON NONLINEAR CONCENTRATION DEPENDENT T-RAYS ABSORPTION

    Directory of Open Access Journals (Sweden)

    YIWEN SUN

    2013-10-01

    Full Text Available T-rays is sensitive to covalently cross-linked proteins and can be used to probe unique dynamic properties of water surrounding a protein. In this paper, we demonstrate the unique absorption properties of the dynamic hydration shells determined by hemagglutinin (HA protein in terahertz frequency. We study the changes arising from different concentrations in detail and show that nonlinear absorption coefficient is induced by the dynamic hydration water. The binary and ternary component model were used to interpret the nonlinearity absorption behaviors and predict the thickness of the hydration shells around the HA protein in aqueous phase.

  3. Determination of chicken and turkey plasma and serum protein concentrations by refractometry and the biuret method.

    Science.gov (United States)

    Andreasen, C B; Latimer, K S; Kircher, I M; Brown, J

    1989-01-01

    Plasma and serum protein concentrations were determined in chickens and turkeys by refractometry (with human and veterinary refractometers) and by the biuret method. Chicken and turkey serum protein values were significantly lower than respective plasma protein values according to both methods. Refractometer readings for both plasma and serum correlated closely with the results of the biuret test (r2 = 0.72 to 0.97). These findings indicate that plasma and serum protein values may be determined accurately in chickens and turkeys with a handheld refractometer.

  4. 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)

  5. Protein digestibility-corrected amino acid scores (PDCAAS) for soy protein isolates and concentrate: criteria for evaluation.

    Science.gov (United States)

    Hughes, Glenna J; Ryan, David J; Mukherjea, Ratna; Schasteen, Charles S

    2011-12-14

    Protein quality, as determined by the PDCAAS method, is a measure of a protein's ability to provide adequate levels of essential amino acids for human needs. PDCAAS is calculated using an amino acid profile and true digestibility of a food protein. Soy protein is recognized as a high quality plant protein, but published PDCAAS values may vary based on the soy protein ingredient as well as the reproducibility and accuracy of the testing methods. Comparison of PDCAAS values for four differently processed soy ingredients, including three isolated soy proteins (ISP) and one soy protein concentrate (SPC), was made using two different laboratories with evaluation of the impact of the reproducibility and accuracy of amino acid profiles. PDCAAS calculations, using amino acid values from one laboratory, yielded a truncated PDCAAS of 1.00 for all four ingredients, while a second laboratory provided statistically significantly lower scores (0.95-1.00). We conclude that analytical method error can be a significant contributor to PDCAAS differences and can be mitigated by the application of amino acid nitrogen recovery correction factors.

  6. Minimum inhibitory concentration of irradiated silk protein powder for bacterial activity

    Energy Technology Data Exchange (ETDEWEB)

    Tuntivisoottikul, Kunya; Bunnak, Jintana [King Mongkut' s Institute of Technology Chaokhun Taharn Ladkrabang, Faculty of Industrial Education, Dept. of Agricultural Educaiton, Bangkok (Thailand); Kume, Tamikazu [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment

    2002-03-01

    The objective of this research was to study a minimum concentration level of irradiated silk protein powder, which inhibited bacterial activity. The concentration of 100 kGy irradiated silk protein powder (ISP) solution was ranged from 5 to 15% in distilled water. The activities of three types of bacteria, Escherichia coli B/r, Bacillus subtilis M3-1 and Staphylococcus aureus K, were tested by using minimum inhibition concentration method (MIC). The results indicated that the minimum concentration level that inhibited growth of E. coli B/r and S. aureus K was 5% ISP and all concentration levels studied could not inhibit the Bacilus subtilis M3-1 activity. (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. Plasma and Pancreas Islet Hormone Concentrations in Lactating Rats Are Associated with Dietary Protein Amounts.

    Science.gov (United States)

    Xu, Lianbin; Lin, Xueyan; White, Robin R; Hanigan, Mark D; Hu, Zhiyong; Hou, Qiuling; Wang, Yun; Wang, Zhonghua

    2018-03-01

    Circulating amino acid (AA) and nitric oxide (NO) concentrations and hepatic gluconeogenesis are affected by previous protein intake. However, information about their relations and islet hormone responses is limited. This study investigated the associations between islet hormone concentrations with circulating AA and NO concentrations as well as with hepatic gluconeogenesis in lactating rats. At delivery, 18 Wistar rats aged 14 wk were assigned either to low-protein (LP; 9% protein), standard-protein (SP; 21% protein), or high-protein (HP; 35% protein) diets for 15 d in groups of 6 pups/dam. Circulating AA and NO concentrations, circulating and pancreas islet hormone concentrations, and the activities and gene expressions of hepatic phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6Pase) were measured at the end of treatment. Circulating insulin and glucagon concentrations were greater in the HP than in the LP (25% and 17%, respectively) and SP (37% and 31%) diet groups, whereas compared with the SP group, pancreatic concentrations were lower in the LP (32% and 49%) and HP (34% and 46%) groups (P < 0.01). Hepatic PEPCK and G6Pase activities in the HP group were greater than those in the SP (15% and 15%) and LP (8% and 19%) groups (P < 0.05). In all groups, plasma NO concentrations were correlated negatively to circulating insulin (r = -0.77, P = 0.0003) and positively to pancreas insulin and glucagon concentrations and the insulin-to-glucagon ratio (r = 0.50-0.63; P < 0.05). Some circulating AAs correlated positively to circulating insulin and pancreas insulin and glucagon (r = 0.50-0.82, P < 0.05) but negatively to circulating glucagon (r = -0.53-0.68, P < 0.05). Variations in circulating AA and NO concentrations and hepatic gluconeogenic enzyme activities are likely intermediary responses involved in the effects of dietary protein amounts on the synthesis and secretion of islet hormones in lactating rats.

  9. 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.

  10. Interrupted hydrogen/deuterium exchange reveals the stable core of the remarkably helical molten globule of alpha-beta parallel protein flavodoxin.

    NARCIS (Netherlands)

    Nabuurs, S.M.; Mierlo, C.P. van

    2010-01-01

    Kinetic intermediates that appear early during protein folding often resemble the relatively stable molten globule intermediates formed by several proteins under mildly denaturing conditions. Molten globules have a substantial amount of secondary structure but lack virtually all tertiary side-chain

  11. Interrupted hydrogen/deuterium exchange reveals the stable core of the remarkably helical molten globule of a-ß parallel protein flavodoxin

    NARCIS (Netherlands)

    Nabuurs, S.M.; Mierlo, van C.P.M.

    2010-01-01

    Kinetic intermediates that appear early during protein folding often resemble the relatively stable molten globule intermediates formed by several proteins under mildly denaturing conditions. Molten globules have a substantial amount of secondary structure but lack virtually all tertiary side-chain

  12. Comparative analysis of the heat stable proteome of radicles of Medicago truncatula seeds during germination identifies late embryogenesis abundant proteins associated with desiccation tolerance

    NARCIS (Netherlands)

    Boudet, J.; Buitink, J.; Hoekstra, F.A.; Rogniaux, H.; Larré, C.; Satour, P.; Leprince, O.

    2006-01-01

    A proteomic analysis was performed on the heat stable protein fraction of imbibed radicles of Medicago truncatula seeds to investigate whether proteins can be identified that are specifically linked to desiccation tolerance (DT). Radicles were compared before and after emergence (2.8 mm long) in

  13. Formation of stable submicron peptide or protein particles by thin film freezing

    Energy Technology Data Exchange (ETDEWEB)

    Johnston, Keith P.; Engstrom, Joshua; Williams, III, Robert O.

    2017-04-18

    The present invention includes compositions and methods for preparing micron-sized or submicron-sized particles by dissolving a water soluble effective ingredient in one or more solvents; spraying or dripping droplets solvent such that the effective ingredient is exposed to a vapor-liquid interface of less than 50, 100, 150, 200, 250, 200, 400 or 500 cm.sup.-1 area/volume to, e.g., increase protein stability; and contacting the droplet with a freezing surface that has a temperature differential of at least 30.degree. C. between the droplet and the surface, wherein the surface freezes the droplet into a thin film with a thickness of less than 500 micrometers and a surface area to volume between 25 to 500 cm.sup.-1.

  14. 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.

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

    Science.gov (United States)

    Rugbjerg, Peter; Knuf, Christoph; Förster, Jochen; Sommer, Morten O A

    2015-12-01

    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 GFP variants, yeast-enhanced GFP, GFP+ and superfolder GFP to yield a sequence-diverged triple GFP molecule 3vGFP with 74-84% internal repeat identity. Unlike a single GFP, the brightness of 3vGFP allowed characterization of a weak promoter in S. cerevisiae. Utilizing 3vGFP, we further engineered a less leaky Cu(2+)-inducible promoter based on CUP1. The basal expression level of the new promoter was approximately 61% below the wild-type CUP1 promoter, thus expanding the absolute range of Cu(2+)-based gene control. The stability of 3vGFP towards direct-repeat recombination was assayed in S. cerevisiae cultured for 25 generations under strong and slightly toxic expression after which only limited reduction in fluorescence was detectable. Such non-recombinogenic GFPs can help quantify intracellular responses operating a low copy number in recombination-prone organisms. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

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

    Science.gov (United States)

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

    2016-07-03

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

  17. 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

  18. Stable expression of silencing-suppressor protein enhances the performance and longevity of an engineered metabolic pathway.

    Science.gov (United States)

    Naim, Fatima; Shrestha, Pushkar; Singh, Surinder P; Waterhouse, Peter M; Wood, Craig C

    2016-06-01

    Transgenic engineering of plants is important in both basic and applied research. However, the expression of a transgene can dwindle over time as the plant's small (s)RNA-guided silencing pathways shut it down. The silencing pathways have evolved as antiviral defence mechanisms, and viruses have co-evolved viral silencing-suppressor proteins (VSPs) to block them. Therefore, VSPs have been routinely used alongside desired transgene constructs to enhance their expression in transient assays. However, constitutive, stable expression of a VSP in a plant usually causes pronounced developmental abnormalities, as their actions interfere with endogenous microRNA-regulated processes, and has largely precluded the use of VSPs as an aid to stable transgene expression. In an attempt to avoid the deleterious effects but obtain the enhancing effect, a number of different VSPs were expressed exclusively in the seeds of Arabidopsis thaliana alongside a three-step transgenic pathway for the synthesis of arachidonic acid (AA), an ω-6 long chain polyunsaturated fatty acid. Results from independent transgenic events, maintained for four generations, showed that the VSP-AA-transformed plants were developmentally normal, apart from minor phenotypes at the cotyledon stage, and could produce 40% more AA than plants transformed with the AA transgene cassette alone. Intriguingly, a geminivirus VSP, V2, was constitutively expressed without causing developmental defects, as it acts on the siRNA amplification step that is not part of the miRNA pathway, and gave strong transgene enhancement. These results demonstrate that VSP expression can be used to protect and enhance stable transgene performance and has significant biotechnological application. © 2015 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

  19. Influence of Biological Agents Effects on Reduction of Ammonia Concentration in Stables of Intensive Farm Animals Breeding

    Directory of Open Access Journals (Sweden)

    Bohuslav Čermák

    2011-05-01

    Full Text Available The living environment distress is connected currently not only with industrial production but also agriculture is biggest producer of toxic gas – ammonia (NH3 .Emissions of that gas originate mainly in the farm animals breeding and generate within storage and handling with farmyard manure, slurry, poultry excrements and litter. Agriculture influences considerably landscape. has impact on basic effect on soil, water and air. In assessing experiment the preparation Biopolym FZT rumen metabolism and N-balance was found positive effects in terms of increased ammonia nitrogen, the number of ciliates and the reduction of N-compounds in feces. Confirmed the impact on the ammonia content in well-ventilated dairy stable. The economic evaluation depends on the exercise price of milk. In the experiments continued.

  20. 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.

  1. 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

  2. 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

  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. Technical Advance: Stable chloroplast transformation in potato: use of green fluorescent protein as a plastid marker.

    Science.gov (United States)

    Sidorov; Kasten; Pang; Hajdukiewicz; Staub; Nehra

    1999-07-01

    We describe here the development of a reproducible plastid transformation system for potato and regeneration of plants with uniformly transformed plastids. Two distinct tobacco-specific plastid vectors, pZS197 (Prrn/aadA/TpsbA) and pMON30125 (Prrn/GFP/Trps16:PpsbA/aadA/TpsbA), designed for integration into the large single copy and inverted repeat regions of the plastid genome, respectively, were bombarded into leaf explants of potato line FL1607. A total of three transgenic lines were selected out of 46 plates bombarded with pZS197 and three transgenic lines out of 104 plates were obtained with pMON30125. Development of a high frequency leaf-based regenera- tion system, a stringent selection scheme and optimization of biolistic transformation protocol were critical for recovery of plastid transformants. Plastid-expressed green fluorescent protein was used as a visual marker for identification of plastid transformants at the early stage of selection and shoot regeneration. The establishment of a plastid transformation system in potato, which has several advantages over routinely used nuclear transformation, offers new possibilities for genetic improvement of this crop.

  5. A Novel, Highly Stable Fold of the Immunoglobulin Binding Domain of Streptococcal Protein G

    Science.gov (United States)

    Gronenborn, Angela M.; Filpula, David R.; Essig, Nina Z.; Achari, Aniruddha; Whitlow, Marc; Wingfield, Paul T.; Marius Clore, G.

    1991-08-01

    The high-resolution three-dimensional structure of a single immunoglobulin binding domain (B1, which comprises 56 residues including the NH_2-terminal Met) of protein G from group G Streptococcus has been determined in solution by nuclear magnetic resonance spectroscopy on the basis of 1058 experimental restraints. The average atomic root-mean-square distribution about the mean coordinate positions is 0.27 angstrom (overset{circ}{mathrm A}) for the backbone atoms, 0.65 overset{circ}{mathrm A} for all atoms, and 0.39 overset{circ}{mathrm A} for atoms excluding disordered surface side chains. The structure has no disulfide bridges and is composed of a four-stranded β sheet, on top of which lies a long helix. The central two strands (β 1 and β 4), comprising the NH_2- and COOH-termini, are parallel, and the outer two strands (β 2 and β 3) are connected by the helix in a +3x crossover. This novel topology (-1, +3x, -1), coupled with an extensive hydrogen-bonding network and a tightly packed and buried hydrophobic core, is probably responsible for the extreme thermal stability of this small domain (reversible melting at 87^circC).

  6. Physico-chemical and thermal properties of chicken myofibrillar protein concentrate (CMPC) mixed with barley bran flour during frozen storage.

    Science.gov (United States)

    Mastanjević, Krešimir; Kovačević, Dragan

    2018-01-01

    Possible new additives for surimi-like products made from chicken meat, which could improve its functional properties during frozen storage, are the subject of much research. The use of dietary fibre in surimi-like products made from chicken meat has not been extensively studied. The objective of this study was to determine the effectiveness of barley bran flour in stabilizing chicken myofibrillar proteins during frozen storage and maintaining its functionality. Surimi-like material (chicken myofibrillar protein concentrate – CMPC) from mechanically deboned chicken meat was mixed with barley bran flour (0–6%) and stored in a freezer for 30, 60, 90 and 180 days. Instrumental color measurements (L*, a*, and b* values) were taken using a Hunter-Lab Mini ScanXE. Texture profile analysis (TPA) tests were performed using a TA.XT2i SMS Stable Micro Systems Texture Analyzer) equipped with an aluminium cylindrical probe P/75. Differential scanning calorim- etry (DSC) was used for the determination of denaturation temperatures and enthalpies. Denaturation enthalpies of CMPC increased when the mass fraction of barley bran was increased (w = 0–6%). Instrumental color parameters (L*, a* and b*) of CMPC gels were significantly (p storage. The increase in denaturation enthalpies and some instrumental textural and color parameters, indicate possible interactions of chicken myofibrillar proteins with barley bran.

  7. 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...

  8. 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.

  9. 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.

  10. Bioactive Whey Protein Concentrate and Lactose Stimulate Gut Function in Formula-fed Preterm Pigs

    DEFF Research Database (Denmark)

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

    2018-01-01

    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-processing are im......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...

  11. 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.

  12. Sensory aroma characteristics of alcalase hydrolyzed rice bran protein concentrate as affected by spray drying and sugar addition

    OpenAIRE

    Arsa, Supeeraya; Theerakulkait, Chockchai

    2014-01-01

    The sensory aroma characteristics of alcalase hydrolyzed rice bran protein concentrate as affected by spray drying and sugar addition were investigated. Rice bran protein concentrate (RBPC) was hydrolyzed by alcalase. Sucrose, glucose or fructose was added to the liquid rice bran protein hydrolysate (LRBPH) and subsequently spray dried. The sensory aroma intensities of the hydrolysates were evaluated. Results showed that after spray drying, the rice bran protein concentrate powder (RBPC-P) ha...

  13. The influence of season and leaf age on concentrations of radiocaesium (137Cs), stable caesium (133Cs) and potassium in Agrostis capillaris

    International Nuclear Information System (INIS)

    Salt, C.A.; Kay, J.W.; Jarvis, K.E.

    2004-01-01

    The transfer of radioactive caesium from soils to plants has been well researched. In contrast there is limited knowledge on natural stable 133 Cs and its potential role as a predictor for radiocaesium behaviour. In a pot experiment with Agrostis capillaris close correlations were found between plant 137 Cs and plant 133 Cs concentrations (R 2 90-96%). Season and leaf age had significant effects with concentrations increasing 10-30-fold between June and December. Simultaneously the plant concentrations of K, the nutrient analogue of Cs, decreased to around one third. In the soil the exchangeable fractions of K and 137 Cs declined. No clear relationships were found between 137+133 Cs in the plant and exchangeable K in the soil. However, at the end of the experiment the K content of the above-ground biomass was higher than the exchangeable pool in the soil, suggesting that depletion of soil K could be a key factor in the observed increase of plant 137+133 Cs over time. - Season and leaf age have the same influence on radiocaesium and stable caesium in Agrostis capillaris but affect K differently

  14. 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.

    2018-03-01

    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.

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

    Science.gov (United States)

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

    2009-10-01

    The objectives of this study were to identify and compare the composition, flavor, and volatile components of serum protein concentrate (SPC) and whey protein concentrate (WPC) containing about 34% protein made from the same milk to each other and to commercial 34% WPC from 6 different factories. The SPC and WPC were manufactured in triplicate with each pair of serum and traditional whey protein manufactured from the same lot of milk. At each replication, SPC and WPC were spray dried (SD) and freeze dried (FD) to determine the effect of the heat used in spray drying on sensory properties. A trained sensory panel documented the sensory profiles of rehydrated SD or FD powders. Volatile components were extracted by solid-phase microextraction (SPME) and solvent extraction followed by solvent-assisted flavor evaporation (SAFE) with gas chromatography-mass spectrometry and gas chromatography-olfactometry. Whey protein concentrates had higher fat content, calcium, and glycomacropeptide content than SPC. Color differences (Hunter L, a, b) were not evident between SPC and WPC powders, but when rehydrated, SPC solutions were clear, whereas WPC solutions were cloudy. No consistent differences were documented in sensory profiles of SD and FD SPC and WPC. The SD WPC had low but distinct buttery (diacetyl) and cardboard flavors, whereas the SD SPC did not. Sensory profiles of both rehydrated SD products were bland and lower in overall aroma and cardboard flavor compared with the commercial WPC. Twenty-nine aroma impact compounds were identified in the SPC and WPC. Lipid and protein oxidation products were present in both products. The SPC and WPC manufactured in this study had lower total volatiles and lower concentrations of many lipid oxidation compounds when compared with commercial WPC. Our results suggest that when SPC and WPC are manufactured under controlled conditions in a similar manner from the same milk using the same ultrafiltration equipment, there are few sensory

  16. 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.

  17. 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.

  18. 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.

  19. 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.

  20. Investigation of whey protein concentration by ultrafiltration elements designed for water treatment

    Directory of Open Access Journals (Sweden)

    Kukučka Miroslav Đ.

    2013-01-01

    Full Text Available Suitability of polysulfone ultrafiltration membranes (UFM commercial designed for water treatment have been investigated for separation of protein (PR from sweet whey. Ultrafiltration (UF of whey originated from dairy has been realized by self-made pilot plant which has been in service about one year. Influence of two whey temperatures (9 oC and 30 oC on efficiency of protein concentration has been examined. Application of investigated UF elements has given whey protein concentrate (WPC with 5 to 6 times excess amount of protein content in regard to starting one. In the same time the prevalent content of lactose has been removed to permeate. Better results have been occurred during the cold whey filtration. Besides the fact that molecular weight cut-off (MWCO of investigated membranes were 50-100 kDa, results showed very successful concentrating of whey proteins of dominantly lower molar weights than 50-100 kDa. Investigated membranes are beneficial for design and construction of UF plants for exploitation in small dairies.

  1. 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...

  2. 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…

  3. 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 ...

  4. 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 ...

  5. 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 ...

  6. Rumen escape protein in concentrate ingredients determined with the nylon bag and enzymatic techniques

    NARCIS (Netherlands)

    Cone, J.W.; Kamman, A.A.; Gelder, A.H. van; Hindle, V.A.

    2002-01-01

    Rumen escape protein (REP) was determined for 26 concentrate feed ingredients using the nylon bag technique and in vitro using a proteolytic enzyme preparation of Streptomyces griseus. In vitro, the samples were incubated for 0, 1, 6 and 24 h. The highest correlation was observed between percentage

  7. 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.

  8. 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

  9. Preparation and physicochemical properties of protein concentrate and isolate produced fromAcacia 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.

  10. Effect of whey concentration on protein recovery in fresh ovine ricotta cheese.

    Science.gov (United States)

    Salvatore, E; Pes, M; Falchi, G; Pagnozzi, D; Furesi, S; Fiori, M; Roggio, T; Addis, M F; Pirisi, A

    2014-01-01

    Ricotta cheese, particularly the ovine type, is a typical Italian dairy product obtained by heat-coagulation of the proteins in whey. The aim of this work was to investigate the influence of whey protein concentration, obtained by ultrafiltration, on yield of fresh ovine ricotta cheese. Ricotta cheeses were obtained by thermocoagulation of mixtures with protein content of 1.56, 3.10, 4.16, and 7.09g/100g from the mixing of skim whey and ultrafiltered skim whey. A fat-to-protein ratio of 1.1 (wt/wt) was obtained for all mixtures by adding fresh cream. The initial mixtures, as well as the final ricotta cheeses, were analyzed for their composition and by SDS-PAGE. Protein bands were quantified by QuantityOne software (Bio-Rad, Hercules, CA) and identified by liquid chromatography-tandem mass spectrometry. Significant differences in the composition of the ricotta cheese were observed depending on protein concentration. Particularly, ricotta cheese resulting from the mixture containing 7.09g/100g of protein presented higher moisture (72.88±1.50g/100g) and protein (10.18±0.45g/100g) contents than that prepared from the mixture with 1.56g/100g of protein (69.52±1.75 and 6.70±0.85g/100g, respectively), and fat content was lower in this sample (12.20±1.60g/100g) compared with the other treatments, with mean values between 15.72 and 20.50g/100g. Each protein fraction presented a different behavior during thermocoagulation. In particular, the recovery of β-lactoglobulin and α-lactalbumin in the cheese increased as their content increased in the mixtures. It was concluded that concentrating ovine rennet whey improved the extent of heat-induced protein aggregation during the thermal coagulation process. This resulted in a better recovery of each protein fraction in the product, and in a consequent increase of ricotta cheese yield. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  11. Immunoglobulin E-reactive proteins in cashew (Anacardium occidentale) apple juice concentrate.

    Science.gov (United States)

    Comstock, Sarah S; Robotham, Jason M; Tawde, Pallavi; Kshirsagar, Harshal; Sathe, Shridhar K; Roux, Kenneth H; Teuber, Suzanne S

    2008-07-23

    Cashew apple juice has the potential to be a natural source of vitamin C and sugar in processed foods. The juice of the cashew apple is obtained by pressing the fleshy peduncle or receptacle, which forms a rounded apple that sits above the true fruit, the cashew nut. Cashew nut allergy is the second most commonly reported tree nut allergy in the United States. To determine if cashew apple juice contains cashew nut allergens, immunoblotting was performed using a cashew apple juice 6X concentrate that was extracted and further concentrated through dialysis, lyophilization, and resuspension. Serum IgE of individuals allergic to cashew nut bound proteins in the cashew apple juice concentrate extract. For some serum samples, IgE reactivity could be inhibited by preincubation of the serum with cashew nut extract, suggesting the presence of cashew nut-related allergens. Using monoclonal antibodies specific for cashew nut allergens, the concentrate was found to contain Ana o 1 (vicilin) and Ana o 2 (legumin). Neither IgE from cashew nut allergic sera nor the monoclonal antibodies bound any peptides in 5 kDa filtered cashew apple juice concentrate. The cashew apple juice concentrate used in these studies contains proteins with IgE-reactive epitopes, including cashew nut legumin and vicilin. No IgE-binding peptides remained after 5 kDa filtration of the concentrate.

  12. A multiple path photonic lab on a chip for parallel protein concentration measurements.

    Science.gov (United States)

    Rodríguez-Ruiz, Isaac; Conejero-Muriel, Mayte; Ackermann, Tobias N; Gavira, José A; Llobera, Andreu

    2015-02-21

    We propose a PDMS-based photonic system for the accurate measurement of protein concentration with minute amounts of the sample. As opposed to the state of the art approach, in the multiple path photonic lab on a chip (MPHIL), analyte concentration or molar absorptivity is obtained with a single injection step, by performing simultaneous parallel optical measurements varying the optical path length. Also, as opposed to the standard calibration protocol, the MPHIL approach does not require a series of measurements at different concentrations. MPHIL has three main advantages: firstly the possibility of dynamically selecting the path length, always working in the absorbance vs. concentration linear range for each target analyte. Secondly, a dramatic reduction of the total volume of the sample required to obtain statistically reliable results. Thirdly, since only one injection is required, the measurement time is minimized, reducing both contamination and signal drifts. These characteristics are clearly advantageous when compared to commercial micro-spectrophotometers. The MPHIL concept was validated by testing three commercial proteins, lysozyme (HEWL), glucose isomerase (d-xylose-ketol-isomerase, GI) and Aspergillus sp. lipase L (BLL), as well as two proteins expressed and purified for this study, B. cereus formamidase (FASE) and dihydropyrimidinase from S. meliloti CECT41 (DHP). The use of MPHIL is also proposed for any spectrophotometric measurement in the UV-VIS range, as well as for its integration as a concentration measurement platform in more advanced photonic lab on a chip systems.

  13. 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)

  14. Pan-Arctic concentrations of mercury and stable isotope ratios of carbon (δ13C) and nitrogen (δ15N) in marine zooplankton

    International Nuclear Information System (INIS)

    Pomerleau, Corinne; Stern, Gary A.; Pućko, Monika; Foster, Karen L.; Macdonald, Robie W.; Fortier, Louis

    2016-01-01

    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 13 C) and nitrogen (δ 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 15 N and lower δ 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.

  15. New procyanidin B3-human salivary protein complexes by mass spectrometry. Effect of salivary protein profile, tannin concentration, and time stability.

    Science.gov (United States)

    Perez-Gregorio, Maria Rosa; Mateus, Nuno; De Freitas, Victor

    2014-10-15

    Several factors could influence the tannin-protein interaction such as the human salivary protein profile, the tannin tested, and the tannin/protein ratio. The goal of this study aims to study the effect of different salivas (A, B, and C) and different tannin concentrations (0.5 and 1 mg/mL) on the interaction process as well as the complex's stability over time. This study is focused on the identification of new procyanidin B3-human salivary protein complexes. Thus, 48 major B3-human salivary protein aggregates were identified regardless of the saliva and tannin concentration tested. A higher number of aggregates was found at lower tannin concentration. Moreover, the number of protein moieties involved in the aggregation process was higher when the tannin concentration was also higher. The selectivity of the different groups of proteins to bind tannin was also confirmed. It was also verified that the B3-human salivary protein complexes formed evolved over time.

  16. Prognosis by C-reactive protein and matrix metalloproteinase-9 levels in stable coronary heart disease during 15 years of follow-up

    DEFF Research Database (Denmark)

    Eldrup, N; Kragelund, C; Steffensen, R

    2012-01-01

    associate with prognosis in patients with stable coronary heart disease. METHODS AND RESULTS: We measured baseline plasma CRP and matrix metalloproteinase-9 in 1090 patients with stable coronary heart disease and as the primary composite endpoint detected incident unstable angina, myocardial infarction......BACKGROUND AND AIM: Elevated CRP and matrix metalloproteinase-9 associate with increased risk of cardiovascular events, possibly because these plasma proteins mark vulnerable atherosclerotic plaques. We tested the hypothesis that levels of C-reactive protein (CRP) and matrix metalloproteinase-9...

  17. 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

    was higher in the VEG diet than in the FM diet (93 versus 92%; t-test, Pb0.05), supporting that protease inhibitors from plant protein ingredients were not the cause of the delay. The apparent digestibility coefficient of carbohydrates (calculated as nitrogen-free extract (NFE)) was much lower in the VEG......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...... 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...

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

    DEFF Research Database (Denmark)

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

    2010-01-01

    Dimeric ligands can be potent inhibitors of protein-protein or enzyme-substrate interactions. They have increased affinity and specificity toward their targets due to their ability to bind two binding sites simultaneously and are therefore attractive in drug design. However, few studies have...... addressed the kinetic mechanism of interaction of such bivalent ligands. We have investigated the binding interaction of a recently identified potent plasma-stable dimeric pentapeptide and PDZ1-2 of postsynaptic density protein-95 (PSD-95) using protein engineering in combination with fluorescence...

  19. 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

  20. Digestible indispensable amino acid score (DIAAS) and protein digestibility corrected amino acid score (PDCAAS) in oat protein concentrate measured in 20- to 30-kilogram pigs.

    Science.gov (United States)

    Abelilla, Jerubella J; Liu, Yanhong; Stein, Hans H

    2018-01-01

    Oat protein concentrate is often used in human food, but the quality of this protein has not been characterized. Therefore, the objectives of this experiment were to determine the standardized ileal digestibility (SID) of crude protein (CP) and amino acids (AA) in oat protein concentrate and to determine differences in protein quality estimates between the protein digestibility-corrected AA score (PDCAAS) and the digestible indispensable AA score (DIAAS) when using growing pigs for both measurements. For infants, the most limiting AA in oat protein concentrate was the aromatic AA (Phe + Tyr), for which the DIAAS value was 41 and the PDCAAS was 43. For children (6 months to 3 years) and children older than 3 years, the most limiting AA in oat protein concentrate was Lys, for which the DIAAS was 56 and 67 and the PDCAAS was 58 and 69, respectively. The DIAAS value for oat protein concentrate was close to the calculated value for PDCAAS, but below the recommended intake for protein. Therefore, to satisfy the daily human AA requirement, oat protein needs to be complemented by other proteins of higher quality and specifically with greater lysine concentrations. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  1. Development of soybeans with low P34 allergen protein concentration for reduced allergenicity of soy foods.

    Science.gov (United States)

    Watanabe, Daisuke; Adányi, Nóra; Takács, Krisztina; Maczó, Anita; Nagy, András; Gelencsér, Éva; Pachner, Martin; Lauter, Kathrin; Baumgartner, Sabine; Vollmann, Johann

    2017-02-01

    In soybean, at least 16 seed proteins have been identified as causing allergenic reactions in sensitive individuals. As a soybean genebank accession low in the immunodominant protein P34 (Gly m Bd 30K) has recently been found, introgression of the low-P34 trait into adapted soybean germplasm has been attempted in order to improve the safety of food products containing soybean protein. Therefore, marker-assisted selection and proteomics were applied to identify and characterize low-P34 soybeans. In low-P34 lines selected from a cross-population, concentrations of the P34 protein as identified with a polyclonal antibody were reduced by 50-70% as compared to P34-containing controls. Using 2D electrophoresis and immunoblotting, the reduction of P34 protein was verified in low-P34 lines. This result was confirmed by liquid chromatographic-tandem mass spectrometric analysis, which revealed either a reduction or complete absence of the authentic P34 protein as suggested from presence or absence of a unique peptide useful for discriminating between conventional and low-P34 lines. Marker-assisted selection proved useful for identifying low-P34 soybean lines for the development of hypoallergenic soy foods. The status of the P34 protein in low-P34 lines needs further characterization. In addition, the food safety relevance of low-P34 soybeans should be tested in clinical studies. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  2. Proteins induced during adaptation of Acetobacter aceti to high acetate concentrations.

    Science.gov (United States)

    Steiner, P; Sauer, U

    2001-12-01

    As a typical product of microbial metabolism, the weak acid acetate is well known for its cytotoxic effects. In contrast to most other microbes, the so-called acetic acid bacteria can acquire significant resistance to high acetate concentrations when properly adapted to such hostile conditions. To characterize the molecular events that are associated with this adaptation, we analyzed global protein expression levels during adaptation of Acetobacter aceti by two-dimensional gel electrophoresis. Adaptation was achieved by using serial batch and continuous cultivations with increasing acetate supplementation. Computer-aided analysis revealed a complex proteome response with at least 50 proteins that are specifically induced by adaptation to acetate but not by other stress conditions, such as heat or oxidative or osmotic stress. Of these proteins, 19 were significantly induced in serial batch and continuous cultures and were thus noted as acetate adaptation proteins (Aaps). Here we present first microsequence information on such Aaps from A. aceti. Membrane-associated processes appear to be of major importance for adaptation, because some of the Aap bear N-terminal sequence homology to membrane proteins and 11 of about 40 resolved proteins from membrane protein-enriched fractions are significantly induced.

  3. In vivo self-assembly of stable green fluorescent protein fusion particles and their uses in enzyme immobilization.

    Science.gov (United States)

    Venning-Slater, Mark; Hooks, David O; Rehm, Bernd H A

    2014-05-01

    Bacterial inclusion bodies are aggregations of mostly inactive and misfolded proteins. However, previously the in vivo self-assembly of green fluorescent protein (GFP) fusions into fluorescent particles which displayed specific binding sites suitable for applications in bioseparation and diagnostics was demonstrated. Here, the suitability of GFP particles for enzyme immobilization was assessed. The enzymes tested were a thermostable α-amylase from Bacillus licheniformis, N-acetyl-d-neuraminic acid aldolase (NanA) from Escherichia coli, and organophosphohydrolase (OpdA) from Agrobacterium radiobacter. Respective GFP particles were isolated and could be stably maintained outside the cell. These enzyme-bearing GFP particles exhibited considerable stability across a range of temperature, pH, and storage conditions and could be recycled. The α-amylase-bearing particles retained activity after treatments at 4 to 85°C and at pHs 4 to 10, were stable for 3 months at 4°C, and could be recycled up to three times. OpdA-bearing particles retained degradation activity after treatments at 4 to 45°C and at pHs 5 to 10 and were able to be recycled up to four times. In contrast, the performance of NanA-bearing particles rapidly declined (>50% loss) after each recycling step and 3 months storage at 4°C. However, they were still able to convert N-acetylmannosamine and pyruvate to N-acetylneuraminic acid after treatment at 4 to 85°C and at pHs 4 to 11. Fluorescent GFP fusion particles represent a novel method for the immobilization and display of enzymes. Potential applications include diagnostic assays, biomass conversion, pharmaceutical production, and bioremediation.

  4. Relationship of serum bilirubin concentration to kidney function and 24-hour urine protein in Korean adults.

    Science.gov (United States)

    Shin, Ho Sik; Jung, Yeon Soon; Rim, Hark

    2011-06-28

    The relationships among serum bilirubin concentration, kidney function and proteinuria have yet to be fully elucidated, nor have these relationships been investigated in Korean adults. We retrospectively reviewed the medical records of Korean adults who were evaluated at Kosin University Gospel Hospital (Busan, Republic of Korea) during a five-year period from January 2005 to December 2009. We evaluated the relationships among serum bilirubin concentration, estimated glomerular filtration rate (eGFR) and 24-hour urinary protein excretion in a sample of 1363 Korean adults aged 18 years or older. The values of eGFR urine albumin ≥150 mg/day were observed in 26.1% (n = 356) and 40.5% (n = 553) of subjects, respectively. Fasting glucose levels ≥126 mg/dL were observed in 44.9% (n = 612) of the total sample. After adjustment for potential confounding factors including demographic characteristics, comorbidities and other laboratory measures, total serum bilirubin was positively associated with eGFR and negatively associated with proteinuria both in the whole cohort and in a subgroup of diabetic individuals. To our knowledge, this is the first hospital-based study specifically aimed at examining the relationships among serum total bilirubin concentration, 24-hour urine protein and kidney function in Korean adults. We demonstrated that serum total bilirubin concentration was negatively correlated with 24-hour urine protein and positively correlated with eGFR in Korean non-diabetic and diabetic adults.

  5. 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.

  6. 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.

  7. 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.

  8. Nutritional, physicochemical, and functional properties of protein concentrate and isolate of newly-developed Bambara groundnut (Vigna subterreneaL.) cultivars.

    Science.gov (United States)

    Adeleke, Olaposi R; Adiamo, Oladipupo Q; Fawale, Olumide S

    2018-01-01

    Bambara groundnut is an indigenous African vegetable grown mainly for human food and animal feed due to its high protein content. Different factors like varieties and origin can influence the chemical composition of Bambara groundnut cultivars. Therefore, the aims of this study are to produce defatted flour and protein concentrate from newly developed Bambara groundnut cultivars [Accessions No: TVSU 5 - Bambara Groundnut White (BGW) and TVSU 146 - Bambara Groundnut Brown (BGB)] and compare their nutritional, physicochemical, and functional properties with market sample [Bambara groundnut commercial (BGC)]. Higher protein content was observed in BGW (20.73%) and BGB (20.14%) as compared to BGC (18.50%). Also, the fat and ash contents of BGB and BGW were higher than that of BGC. Also, the new varieties were found to contain higher levels of some essential fatty acids such as linoleic and linolenic acids. The concentration of thiamine, riboflavin, niacin, pantothenic, ascorbic acids, pyrodoxine, alpha tocopherol, and vitamin K were also significantly higher in the two new varieties. The new varieties were good sources of magnesium, calcium, iron, manganese, sodium, and potassium. The oil and water absorption and swelling capacities of whole, defatted, and protein concentrate flour of the new varieties increase with increase in temperature. The defatted flour and protein concentrate of brown Bambara groundnut was found to exhibit high emulsifying activity and stability at different pH's and salt concentrations. The new varieties possess significantly higher foaming capacity and stability than the commercial variety. The results obtained from this study have shown the potential for the industrial and household use of the new Bambara groundnut cultivars into shelf stable protein products and could be a useful ingredient in food formulations.

  9. 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.

  10. 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.

  11. 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

  12. Effects of salt concentration on the reaction rate of Glc with amino acids, peptides, and proteins.

    Science.gov (United States)

    Yamaguchi, Keiko; Noumi, Yuri; Nakajima, Katsumi; Nagatsuka, Chiharu; Aizawa, Haruko; Nakawaki, Rie; Mizude, Eri; Otsuka, Yuzuru; Homma, Takeshi; Chuyen, Nguyen Van

    2009-11-01

    The reaction between the amino group and the carbonyl group is important in food quality control. Furthermore, advanced glycation end products from foods are considered to relate to aging and diabetes. Thus, it is important to control this reaction. In this study, we investigated the effects of salt concentration on the rates of browning reaction of amino acid, peptides, and proteins. A high concentration of sodium chloride retarded the reaction rate of Glc with amino acids as measured with the absorbance at 470 nm, but did not change the browning rate of Glc with peptides. On the other hand, sodium chloride retarded the browning reaction rate of proteins as measured with polymerization degree or by the loss of Lys. It is hoped that the results of this study will be applied in the control of amino-carbonyl reaction rates in the food industry.

  13. Dosimetry Determines the Initial OH Radical Concentration in Fast Photochemical Oxidation of Proteins (FPOP)

    Science.gov (United States)

    Niu, Ben; Zhang, Hao; Giblin, Daryl; Rempel, Don L.; Gross, Michael L.

    2015-05-01

    Fast photochemical oxidation of proteins (FPOP) employs laser photolysis of hydrogen peroxide to give OH radicals that label amino acid side-chains of proteins on the microsecond time scale. A method for quantitation of hydroxyl radicals after laser photolysis is of importance to FPOP because it establishes a means to adjust the yield of •OH, offers the opportunity of tunable modifications, and provides a basis for kinetic measurements. The initial concentration of OH radicals has yet to be measured experimentally. We report here an approach using isotope dilution gas chromatography/mass spectrometry (GC/MS) to determine quantitatively the initial •OH concentration (we found ~0.95 mM from 15 mM H2O2) from laser photolysis and to investigate the quenching efficiencies for various •OH scavengers.

  14. Human Cytomegalovirus gH/gL Forms a Stable Complex with the Fusion Protein gB in Virions.

    Directory of Open Access Journals (Sweden)

    Adam L Vanarsdall

    2016-04-01

    Full Text Available Human cytomegalovirus (HCMV is a ubiquitous virus that is a major pathogen in newborns and immunocompromised or immunosuppressed patients. HCMV infects a wide variety of cell types using distinct entry pathways that involve different forms of the gH/gL glycoprotein: gH/gL/gO and gH/gL/UL128-131 as well as the viral fusion glycoprotein, gB. However, the minimal or core fusion machinery (sufficient for cell-cell fusion is just gH/gL and gB. Here, we demonstrate that HCMV gB and gH/gL form a stable complex early after their synthesis and in the absence of other viral proteins. gH/gL can interact with gB mutants that are unable to mediate cell-cell fusion. gB-gH/gL complexes included as much as 16-50% of the total gH/gL in HCMV virus particles. In contrast, only small amounts of gH/gL/gO and gH/gL/UL128-131 complexes were found associated with gB. All herpesviruses express gB and gH/gL molecules and most models describing herpesvirus entry suggest that gH/gL interacts with gB to mediate membrane fusion, although there is no direct evidence for this. For herpes simplex virus (HSV-1 it has been suggested that after receptor binding gH/gL binds to gB either just before, or coincident with membrane fusion. Therefore, our results have major implications for these models, demonstrating that HCMV gB and gH/gL forms stable gB-gH/gL complexes that are incorporated virions without receptor binding or membrane fusion. Moreover, our data is the best support to date for the proposal that gH/gL interacts with gB.

  15. 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

  16. Protein resistance efficacy of PEO-silane amphiphiles: Dependence on PEO-segment length and concentration.

    Science.gov (United States)

    Rufin, Marc A; Barry, Mikayla E; Adair, Paige A; Hawkins, Melissa L; Raymond, Jeffery E; Grunlan, Melissa A

    2016-09-01

    In contrast to modification with conventional PEO-silanes (i.e. no siloxane tether), silicones with dramatically enhanced protein resistance have been previously achieved via bulk-modification with poly(ethylene oxide) (PEO)-silane amphiphiles α-(EtO)3Si(CH2)2-oligodimethylsiloxane13-block-PEOn-OCH3 when n=8 and 16 but not when n=3. In this work, their efficacy was evaluated in terms of optimal PEO-segment length and minimum concentration required in silicone. For each PEO-silane amphiphile (n=3, 8, and 16), five concentrations (5, 10, 25, 50, and 100μmol per 1g silicone) were evaluated. Efficacy was quantified in terms of the modified silicones' abilities to undergo rapid, water-driven surface restructuring to form hydrophilic surfaces as well as resistance to fibrinogen adsorption. Only n=8 and 16 were effective, with a lower minimum concentration in silicone required for n=8 (10μmol per 1g silicone) versus n=16 (25μmol per 1g silicone). Silicone is commonly used for implantable medical devices, but its hydrophobic surface promotes protein adsorption which leads to thrombosis and infection. Typical methods to incorporate poly(ethylene oxide) (PEO) into silicones have not been effective due to the poor migration of PEO to the surface-biological interface. In this work, PEO-silane amphiphiles - comprised of a siloxane tether (m=13) and variable PEO segment lengths (n=3, 8, 16) - were blended into silicone to improve its protein resistance. The efficacy of the amphiphiles was determined to be dependent on PEO length. With the intermediate PEO length (n=8), water-driven surface restructuring and resulting protein resistance was achieved with a concentration of only 1.7wt%. Copyright © 2016 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

  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. 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.

  19. 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.

  20. Formation of new stable pigments from condensation reaction between malvidin 3-glucoside and (-)-epicatechin mediated by acetaldehyde: Effect of tartaric acid concentration.

    Science.gov (United States)

    Sun, Baoshan; Barradas, Tania; Leandro, Conceição; Santos, Cláudia; Spranger, Isabel

    2008-09-15

    The objective of this work was to study the effect of tartaric acid concentration on the condensation reaction between malvidin 3-glucoside (Mv-glc) and flavanols mediated by acetaldehyde in the model solution. The model wine solutions were prepared by 12% ethanol in water (v/v) with two different l-tartaric acid concentrations (5g/l and 25g/l, respectively) and at two different pH values (3.2 and 1.7, respectively). Four new pigments were detected in model wine solutions containing Mv-glc, (-)-epicatechin and acetaldehyde. By reverse-phase HPLC-DAD, ESI-MS and MS(n) fragmentation analysis, the four new pigments were tentatively identified as four isomers of hydroxyethyl malvidin-3-glucoside-ethyl-flavanol. The decrease in the concentration of Mv-glc and (-)-epicatechin and the increase in the concentration of the new identified pigments were more pronounced at higher tartaric acid concentration. At pH 1.7, although the two well-recognized ethyl-linked Mv-glc-flavanol isomers were quantitatively the major pigmented products in the reaction solution throughout the assay period, they appeared less stable than the four new pigments. At pH 3.2, the rate of formation of ethyl-linked Mv-glc-flavanol pigments was much slower than at pH 1.7, whereas the four new pigments were quantitatively the predominant pigmented products at the latter stage of the reaction. Copyright © 2008 Elsevier Ltd. All rights reserved.

  1. Passive dosing of triclosan in multigeneration tests with copepods - stable exposure concentrations and effects at the low μg/L range.

    Science.gov (United States)

    Ribbenstedt, Anton; Mustajärvi, Lukas; Breitholtz, Magnus; Gorokhova, Elena; Mayer, Philipp; Sobek, Anna

    2017-05-01

    Ecotoxicity testing is a crucial component of chemical risk assessment. Still, due to methodological difficulties related to controlling exposure concentrations over time, data on long-term effects of organic chemicals at low concentrations are limited. The aim of the present study was, therefore, to test the applicability of passive dosing to maintain stable concentrations of the organochlorine bacteriocide triclosan in the water phase during a 6-wk multigeneration 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 (D silicone-water ) was 10466 ± 1927. A population development test was conducted at 3 concentration levels of triclosan that were measured to be 3 μg/L to 5 μg/L, 7 μg/L to 11 μg/L and 16 μg/L to 26 μg/L. The results demonstrate that passive dosing is applicable for long-term ecotoxicity testing of organic chemicals, including during significant growth of the test organism population. Shifts in the demographic structure of the population during exposure suggest the most severe effects were exerted on juvenile development. Progressively lower development index values in the populations exposed to increasing triclosan concentrations suggest developmental retardation. The results further stress the need for chronic exposure during ecotoxicity testing in chemical risk assessment because even the most sensitive endpoint was not significant until after 7 d of exposure. Environ Toxicol Chem 2017;36:1254-1260. © 2016 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC. © 2016 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals, Inc. on behalf of SETAC.

  2. 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.

  3. Determination of the sulfur and oxygen stable isotopes concentration by mass spectrometry; Determinacao da concentracao (atomos %) dos isotopos estaveis de enxofre e oxigenio por espectrometria de massas

    Energy Technology Data Exchange (ETDEWEB)

    Bendassolli, Jose A.; Trivelin, Paulo C.O. [Centro de Energia Nuclear na Agricultura (CENA), Piracicaba, SP (Brazil); Carneiro Junior, Francisco [Universidade Metodista de Piracicaba (UNIMEP), SP (Brazil)

    1997-10-01

    Two methods to determine the isotope concentrations of {sup 32} S, {sup 33} S, {sup 34} S, {sup 36} S and {sup 18} O in SO{sub 2} samples were evaluated. The isotope analysis were carried out in an C H{sub 4} model, ATLAS MAT mass spectrometer, through the electromagnetic mass scanning of the mass regions 48, 49, 50, 51 and 52. For assessing the effect of {sup 18} O contribution in the determination of the {sup 34} S isotope concentration, two calculation procedures were used. The first involved the resolution of a system of six equations with six unknown quantities, where the values in atoms % for all stable isotopes ({sup 32} S, {sup 33} S, {sup 34} S and {sup 36} S) as well as the most abundant oxygen isotopes ({sup 16} O and {sup 18} O) in SO{sub 2} samples were obtained. The second procedure used a simplified first degree equation, where the O{sub 2} natural abundance was always assumed, to calculate the {sup 34} S concentration only. (author). 11 refs., 2 figs., 3 tabs.

  4. Trends in nitrate concentrations and determination of its origin using stable isotopes (18O and 15N) in groundwater of the Western Central Valley, Costa Rica.

    Science.gov (United States)

    Reynolds-Vargas, Jenny; Fraile-Merino, Julio; Hirata, Ricardo

    2006-08-01

    A study was conducted to evaluate long-term trends in nitrate concentrations and to try to identify the origin of nitrate using stable isotopes (15N(NO3-) and 18O(NO3-)) in the aquifers of the western Central Valley, Costa Rica, where more than 1 million people depend on groundwater to satisfy their daily needs. Data from 20 sites periodically sampled for 4 to 17 years indicate an increasing trend in nitrate concentrations at five sites, which in a period ranging from 10 to 40 years, will exceed recommended maximum concentrations. Results of isotopic analysis indicate a correspondence between land use patterns and the isotopic signature of nitrate in groundwater and suggest that urbanization processes without adequate waste disposal systems, followed by coffee fertilization practices, are threatening water quality in the region. We conclude that groundwater management in this area is not sustainable, and that land use substitution processes from agricultural activity to residential occupation that do not have proper sewage disposal systems may cause a significant increment in the nitrate contaminant load.

  5. 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.

  6. 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.

  7. Short communication: Effects of nanofiltration and evaporation on the physiochemical properties of milk protein during processing of milk protein concentrate.

    Science.gov (United States)

    Cao, Jialu; Zhang, Wei; Wu, Shaozong; Liu, Chang; Li, Yan; Li, Haimei; Zhang, Liebing

    2015-01-01

    The aim of this work was to evaluate the effects of nanofiltration and evaporation concentration technologies on the physiochemical properties of milk protein concentrate (MPC) during processing. Skim milk, ultrafiltered milk, evaporated milk, nanofiltered milk, evaporated MPC, and nanofiltered MPC samples were collected at different processing stages. Chemical composition, microstructure of casein micelles, free sulfhydryl content, and surface hydrophobicity of the samples were determined. The insolubility index of MPC was also determined. Casein micelles aggregated compactly after evaporation while surface hydrophobicity increased and free sulfhydryl content decreased in evaporated milk compared with skim milk. However, the microstructure of the casein micelles was relatively undisturbed after nanofiltration, with reduced surface hydrophobicity and free sulfhydryl content. No significant difference was found in chemical composition between the 2 MPC preparations: approximately 61.40% protein and 28.49% lactose. In addition, the particulate microstructures of both MPC were similar. However, the insolubility index of evaporated MPC was significantly (0.58mL) higher than that of nanofiltered MPC. Nanofiltration may be an effective way to improve the solubility of MPC products. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  8. 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...... 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...... and MBL and by use of density gradient ultra centrifugation we could show that the major part of serum MAP-1 circulates in complex with the LCP molecules. In conclusion, these results show that MAP-1 is a highly stable glycosylated human serum protein found in complex with Ficolin-3, Ficolin-2 and MBL....

  9. 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...

  10. Highly water-soluble ruthenium(II terpyridine coordination compounds form stable adducts with blood-borne metal transporting proteins

    Directory of Open Access Journals (Sweden)

    Marija Nišavić

    2018-03-01

    Full Text Available Three coordination compounds of ruthenium(II, belonging to a recently synthesised series of water-soluble compounds of general formula mer-[Ru(L3(N-NCl]Cl, where L3 = 4'-chloro-2,2':6',2″-terpyridine (Cl-tpy, N-N = ethylenediamine (en, 1,2-diaminocyclohexane (dach or 2,2'-bipyridine (bpy, have shown strong binding to calf thymus DNA and moderate in vitro cytotoxicity towards cancer cell lines. Knowing that serum proteins play a crucial role in the transport and deactivation of ruthenium drugs, we have conducted a detailed study of their interactions with two major metal-transporting serum proteins, albumin and transferrin, and it is presented herein. Ruthenated protein adducts were formed with various concentrations of the three compounds and then separated from the unbound portions by ultrafiltration through 10 kDa cut-off centrifugal filter units. The stoichiometry of binding was determined using inductively coupled plasma optical emission spectrometry. One mol of albumin bound up to 7, 8.5 and 1.5 mol of compound 1 ([Ru(Cl-tpy(enCl][Cl], 2 ([Ru(Cl-tpy(dachCl][Cl] and 3 ([Ru(Cl-tpy(bpyCl][Cl], respectively. One mol of transferrin bound up to 3, 3.5 and 0.4 mol of 1, 2 and 3, respectively. The affinity of albumin and transferrin for the three ruthenium compounds was evaluated using fluorescence quenching. The binding constants for 1 and 2 lay within the range 104–105 M−1, suggesting moderate-to-strong attachment to albumin. Both compounds showed much lower affinity for transferrin (102–103 M−1. Compound 3 bound weakly to each studied protein. High resolution ESI qTOF mass spectra of albumin before and after binding of 1 revealed the high stoichiometry of binding. Although the binding of the compounds 1–3 to albumin and transferrin did not affect proteins’ secondary structure much, their tertiary structures underwent some alterations, as deduced from the circular dichroism study. Changes in the stability of albumin, after

  11. Effect of annatto addition and bleaching treatments on ultrafiltration flux during production of 80% whey protein concentrate and 80% serum protein concentrate.

    Science.gov (United States)

    Adams, Michael C; Zulewska, Justyna; Barbano, David M

    2013-04-01

    The goals of this study were to determine if adding annatto color to milk or applying a bleaching process to whey or microfiltration (MF) permeate influenced ultrafiltration (UF) flux, diafiltration (DF) flux, or membrane fouling during production of 80% whey protein concentrate (WPC80) or 80% serum protein concentrate (SPC80). Separated Cheddar cheese whey (18 vats using 900 kg of whole milk each) and MF permeate of skim milk (18 processing runs using 800 kg of skim milk each) were produced to make WPC80 and SPC80, respectively. The 6 treatments, replicated 3 times each, that constituted the 18 processing runs within either whey or MF permeate UF were as follows: (1) no annatto; (2) no annatto+benzoyl peroxide (BPO); (3) no annatto+hydrogen peroxide (H2O2); (4) annatto; (5) annatto+BPO; and (6) annatto+H2O2. Approximately 700 kg of whey or 530 kg of MF permeate from each treatment were heated to 50°C and processed in 2 stages (UF and DF) with the UF system in batch recirculation mode using a polyethersulfone spiral-wound UF membrane with a molecular weight cutoff of 10,000 Da. Addition of annatto color had no effect on UF or DF flux. The processes of bleaching whey or MF permeate with or without added color improved flux during processing. Bleaching with H2O2 usually produced higher flux than bleaching with BPO. Bleaching with BPO increased WPC80 flux to a greater extent than it did SPC80 flux. Though no differences in mean flux were observed for a common bleaching treatment between the WPC80 and SPC80 production processes during the UF stage, mean flux during WPC80 DF was higher than mean flux during SPC80 DF for each bleaching treatment. Water flux values before and after processing were used to calculate a fouling coefficient that demonstrated differences in fouling which were consistent with flux differences among treatments. In both processes, bleaching with H2O2 led to the largest reduction in fouling. No effect of annatto on fouling was observed. The

  12. 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.

  13. Relationship of serum bilirubin concentration to kidney function and 24-hour urine protein in Korean adults

    Directory of Open Access Journals (Sweden)

    Jung Yeon Soon

    2011-06-01

    Full Text Available Abstract Background The relationships among serum bilirubin concentration, kidney function and proteinuria have yet to be fully elucidated, nor have these relationships been investigated in Korean adults. Method We retrospectively reviewed the medical records of Korean adults who were evaluated at Kosin University Gospel Hospital (Busan, Republic of Korea during a five-year period from January 2005 to December 2009. We evaluated the relationships among serum bilirubin concentration, estimated glomerular filtration rate (eGFR and 24-hour urinary protein excretion in a sample of 1363 Korean adults aged 18 years or older. Results The values of eGFR 2 and 24-hour urine albumin ≥150 mg/day were observed in 26.1% (n = 356 and 40.5% (n = 553 of subjects, respectively. Fasting glucose levels ≥126 mg/dL were observed in 44.9% (n = 612 of the total sample. After adjustment for potential confounding factors including demographic characteristics, comorbidities and other laboratory measures, total serum bilirubin was positively associated with eGFR and negatively associated with proteinuria both in the whole cohort and in a subgroup of diabetic individuals. Conclusions To our knowledge, this is the first hospital-based study specifically aimed at examining the relationships among serum total bilirubin concentration, 24-hour urine protein and kidney function in Korean adults. We demonstrated that serum total bilirubin concentration was negatively correlated with 24-hour urine protein and positively correlated with eGFR in Korean non-diabetic and diabetic adults.

  14. 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...... outcomes were all-cause mortality and myocardial infarction. RESULTS: Patients (n=4243) were followed for a median of 2.8 years. In a Cox analysis, elevated PAPP-A was significantly related to the composite outcome of myocardial infarction and death (HR 1.99, 95% CI 1.62-2.45, p...

  15. 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...

  16. 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.

  17. 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.

  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. 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.

  20. [Dynamic effect of curcumin on urine concentration of neuronal thread protein in AD mice].

    Science.gov (United States)

    Chen, Xiao-Pei; Li, Rui-Sheng; Wang, Hong; Ren, Ying; Sun, Hai-Yun; Yang, Jin-Duo; Wang, Peng-Wen

    2013-05-01

    Through the dynamic detection of the concentration change of the urine Alzheimer-associated neuronal thread protein (AD7C-NTP) in the curcumin treated Alzheimer's disease (AD) model (APP/PS1 double transgenic) mice, the therapeutic effect of curcumin in AD was determined. Thirty three-month-old APP /PS1 double transgenic mice were randomly divided into 5 groups, 6 in each group, the model group, rosiglitazone group(10 mg . kg-1 . d-1) , high(400 mg . kg -1 . d-1) , medium(200 mg . kg-1. d-1) and low(100 mg . kg-1 . d-1) dose curcumin groups. Six C57BL/6J mice in the same age and genetic background were used as normal control group. All the 6 groups of mice were intragastrically administered for 6 months. Urine samples were collected on 4 month, 5 month and 6 month after intragastric administration, respectively. The changes of urinary AD7C-NTP concentration were detected by enzyme-linked immunosorbent assay (ELISA). The concentration of AD7C-NTP of each group was compared at the same time point, the concentration of model group is higher than normal control group (P curcumin dose group with 4 months treatment, has no statistical difference compared with model group. The AD7C-NTP concentration of each group was elevated with the age growth, and all concentrations of the treatment groups were lower than the model group at the same period. With the treatment of 4, 5 and 6 months, the concentration of the normal control group has significant difference with the treatment groups(P curcumin from traditional Chinese medicine can delay the progression of AD.

  1. 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.

  2. GLUCOSE, TOTAL PROTEINS, URIC ACID AND TRIGL YCERIDES CONCENTRATIONS IN BLOOD OF ATIVE LAYING HENS

    Directory of Open Access Journals (Sweden)

    Bashir Mahmood Bhatti, Tanzeela Talat and Rozina Sardar

    2001-09-01

    Full Text Available This study was undertaken, to estimate levels of glucose, total proteins, albumins, uric acid and triglycerides in plasma of Desi and Naked Neck laying hens. The experimental birds received ration containing 16 per cent crude protein and were housed in open sheds. The mean values (mg/dL observed in Desi hens were 226.736+15.20 glucose, 1.624+0.224 albumin, 5.203+1.078 total proteins, 4.633+1.875 uric acid and 529.800+554.74 triglycerides. In Naked neck hens, the mean values were found to be 231.818+31.376 glucose, 1.562+0.287 albumins, 4.533+0.797total proteins, 4.157+1.336 uric acid and 791.200+320.474 triglycerides. There was no difference (P<0.05 in mean values of blood parameters between both the native laying hens which suggested that in identical genetical mechanism regulated concentrations of blood chemical constituents under study.

  3. Development, chemical and sensory characterization of semi skim yogurt added with whey protein concentrate

    Directory of Open Access Journals (Sweden)

    Andressa Regina Antunes

    2015-04-01

    Full Text Available The aim of this work was to develop, to perform the chemical characterization and verify the acceptability of a semi skim yogurt added with whey protein concentrate (WPC 35 %, and compare it to traditional yogurt, without adding any protein source. For this, two formulations were prepared: 1- traditional semi skim and 2: semi skim added with WPC 35 %, with subsequent evaluation of physicochemical characteristics of acidity, pH and viscosity, as well as the compositional analysis of the levels of humidity, full solids, ash, protein, fat and carbohydrates for both formulations. The sensory analysis was performed with 80 untrained tasters, which was based on parameters of appearance, taste, flavor and consistency. The appearance and consistency parameters showed a significant difference between received grades, and the yogurt added with WPC 35 % the most preferred these aspects, as well as the formulation with the highest widespread acceptance among testers. The results indicate that the use of WPC 35 % in foods is a promising alternative, considering the overall acceptance obtained, the protein content beneficial to the consumers health, the improvement of technological properties provided to food coupled to the extent which it is produced, low cost and reducing the damage caused to the environment. 

  4. The Quality of Dory Fillets based on Water Soluble Protein, Color, and Myoglobin Concentration

    Directory of Open Access Journals (Sweden)

    Nurfajrin Nisa

    2016-04-01

    Full Text Available Fillet of dory is very easy to be find in Indonesian market with various brand and produsen.Imported dory fillet is preferred by consumer so far because it has a white color compare than localfillets. Color is the important parameter that used by consumers to determine the quality of filet. Thisstudy was aimed to determine the quality of local and imported fillets, including protein profile usingSDS PAGE, color measurement, and myoglobin extractability. The results of water soluble protein profilesshowed dory fillet contained 13-15 bands. The redness value (a* of local fillet (DN, DL, DM was highercompared others. However, imported fillet (DI had the highest if redness index (a/b. Imported fillet (DIshowed the lowest concentration of myoglobin compared other samples.

  5. The Quality of Dory Fillets based on Water Soluble Protein, Color, and Myoglobin Concentration

    Directory of Open Access Journals (Sweden)

    Nurfajrin Nisa

    2016-04-01

    Full Text Available Fillet of dory is very easy to be find in Indonesian market with various brand and produsen. Imported dory fillet is preferred by consumer so far because it has a white color compare than local fillets. Color is the important parameter that used by consumers to determine the quality of filet. This study was aimed to determine the quality of local and imported fillets, including protein profile using SDS PAGE, color measurement, and myoglobin extractability. The results of water soluble protein profiles showed dory fillet contained 13-15 bands. The redness value (a* of local fillet (DN, DL, DM was higher compared others. However, imported fillet (DI had the highest if redness index (a/b. Imported fillet (DI showed the lowest concentration of myoglobin compared other samples.

  6. MRS thermometry calibration at 3 T: effects of protein, ionic concentration and magnetic field strength.

    Science.gov (United States)

    Babourina-Brooks, Ben; Simpson, Robert; Arvanitis, Theodoros N; Machin, Graham; Peet, Andrew C; Davies, Nigel P

    2015-07-01

    MRS thermometry has been utilized to measure temperature changes in the brain, which may aid in the diagnosis of brain trauma and tumours. However, the temperature calibration of the technique has been shown to be sensitive to non-temperature-based factors, which may provide unique information on the tissue microenvironment if the mechanisms can be further understood. The focus of this study was to investigate the effects of varied protein content on the calibration of MRS thermometry at 3 T, which has not been thoroughly explored in the literature. The effects of ionic concentration and magnetic field strength were also considered. Temperature reference materials were controlled by water circulation and freezing organic fixed-point compounds (diphenyl ether and ethylene carbonate) stable to within 0.2 °C. The temperature was measured throughout the scan time with a fluoro-optic probe, with an uncertainty of 0.16 °C. The probe was calibrated at the National Physical Laboratory (NPL) with traceability to the International Temperature Scale 1990 (ITS-90). MRS thermometry measures were based on single-voxel spectroscopy chemical shift differences between water and N-acetylaspartate (NAA), Δ(H20-NAA), using a Philips Achieva 3 T scanner. Six different phantom solutions with varying protein or ionic concentration, simulating potential tissue differences, were investigated within a temperature range of 21-42 °C. Results were compared with a similar study performed at 1.5 T to observe the effect of field strengths. Temperature calibration curves were plotted to convert Δ(H20-NAA) to apparent temperature. The apparent temperature changed by -0.2 °C/% of bovine serum albumin (BSA) and a trend of 0.5 °C/50 mM ionic concentration was observed. Differences in the calibration coefficients for the 10% BSA solution were seen in this study at 3 T compared with a study at 1.5 T. MRS thermometry may be utilized to measure temperature and the tissue

  7. The effect of starter culture and annatto on the flavor and functionality of whey protein concentrate.

    Science.gov (United States)

    Campbell, R E; Miracle, R E; Drake, M A

    2011-03-01

    The flavor of whey protein can carry over into ingredient applications and negatively influence consumer acceptance. Understanding sources of flavors in whey protein is crucial to minimize flavor. The objective of this study was to evaluate the effect of annatto color and starter culture on the flavor and functionality of whey protein concentrate (WPC). Cheddar cheese whey with and without annatto (15 mL of annatto/454 kg of milk, annatto with 3% wt/vol norbixin content) was manufactured using a mesophilic lactic starter culture or by addition of lactic acid and rennet (rennet set). Pasteurized fat-separated whey was then ultrafiltered and spray dried into WPC. The experiment was replicated 4 times. Flavor of liquid wheys and WPC were evaluated by sensory and instrumental volatile analyses. In addition to flavor evaluations on WPC, color analysis (Hunter Lab and norbixin extraction) and functionality tests (solubility and heat stability) also were performed. Both main effects (annatto, starter) and interactions were investigated. No differences in sensory properties or functionality were observed among WPC. Lipid oxidation compounds were higher in WPC manufactured from whey with starter culture compared with WPC from rennet-set whey. The WPC with annatto had higher concentrations of p-xylene, diacetyl, pentanal, and decanal compared with WPC without annatto. Interactions were observed between starter and annatto for hexanal, suggesting that annatto may have an antioxidant effect when present in whey made with starter culture. Results suggest that annatto has a no effect on whey protein flavor, but that the starter culture has a large influence on the oxidative stability of whey. Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  8. Instrumental and Sensory Texture Attributes of High-Protein Nutrition Bars Formulated with Extruded Milk Protein Concentrate.

    Science.gov (United States)

    Banach, J C; Clark, S; Lamsal, B P

    2016-05-01

    Previous instrumental study of high-protein nutrition (HPN) bars formulated with extruded milk protein concentrate (MPC) indicated slower hardening compared to bars formulated with unmodified MPC. However, hardness, and its change during storage, insufficiently characterizes HPN bar texture. In this study, MPC80 was extruded at 2 different conditions and model HPN bars were prepared. A trained sensory panel and instrumental techniques were used to measure HPN bar firmness, crumbliness, fracturability, hardness, cohesiveness, and other attributes to characterize texture change during storage. Extrusion modification, storage temperature, and storage time significantly affected the instrumental and sensory panel measured texture attributes. The HPN bars became firmer and less cohesive during storage. When evaluated at the same storage conditions, the texture attributes of the HPN bars formulated with the different extrudates did not differ significantly from each other. However, textural differences were noted most of the time between the control and the HPN bars formulated with extruded MPC80. An adapted HPN bar crumbliness measurement technique produced results that were correlated with sensory panel measured crumbliness (r = 0.85) and cohesiveness (r = -0.84). Overall, the HPN bars formulated with extruded MPC80 were significantly softer, less crumbly, and more cohesive than the control during storage. © 2016 Institute of Food Technologists®

  9. Particle Size of Milk Protein Concentrate Powder Affects the Texture of High-Protein Nutrition Bars During Storage.

    Science.gov (United States)

    Banach, J C; Clark, S; Lamsal, B P

    2017-04-01

    Milk protein concentrate powder with 85% protein (MPC85) was jet-milled to give 2 particle size distributions (that is, JM-Coarse and JM-Fine) or freeze-dried (FD), in order to improve the functional properties of MPC85 for use in high-protein nutrition (HPN) bars. Volume-weighted mean diameter decreased from 86 μm to 49, 22, and 8 μm in FD, JM-Coarse, and JM-Fine, respectively (P powders modified by jet-milling and freeze-drying were significantly denser than the control MPC85 (P powders decreased (P < 0.05) by an order of magnitude, yet only FD possessed a lower volume of interstitial air (P < 0.05). Particle size reduction and freeze-drying MPC85 decreased its water holding capacity and improved its dispersibility by at least 20%. Contact angle measurements showed that these modifications increased initial hydrophobicity and did not improve wettability. HPN bars made from JM-Fine or FD were firmer by 40 or 17 N, respectively, than the control on day 0 (P < 0.05). HPN bar maximum compressive force increased by 38%, 33%, and 242% after 42 d at 32 °C when formulated with JM-Fine, FD, or control MPC85, respectively. HPN bars prepared with JM-Fine were less crumbly than those formulated with control or FD MPC85. Physically altering the particle structure of MPC85 improved its ability to plasticize within HPN bars and this improved their cohesiveness and textural stability. © 2017 Institute of Food Technologists®.

  10. 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

  11. 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 .

  12. Effect of protein and glycerol concentration on the mechanical, optical, and water vapor barrier properties of canola protein isolate-based edible films.

    Science.gov (United States)

    Chang, Chang; Nickerson, Michael T

    2015-01-01

    Biodegradable edible films prepared using proteins are both economically and environmentally important to the food packaging industry relative to traditional petroleum-derived synthetic materials. In the present study, the mechanical and water vapor barrier properties of casted canola protein isolate edible films were investigated as a function of protein (5.0% and 7.5%) and glycerol (30%, 35%, 40%, 45%, and 50%) content. Specifically, tensile strength and elongation, elastic modulus, puncture strength and deformation, opacity, and water vapor permeability were measured. Results indicated that tensile strength, puncture strength, and elastic modulus decreased, while tensile elongation and puncture deformation values increased as glycerol concentration increased for both 5.0% and 7.5% canola protein isolate films. Furthermore, tensile strength, puncture strength, and elastic modulus values were found to increase at higher protein concentrations within the canola protein isolate films, whereas puncture deformation values decreased. Tensile elongation was found to be similar for both canola protein isolate protein levels. Canola protein isolate films became more transparent with increasing of glycerol concentration and decreasing of canola protein isolate concentration. Water vapor permeability value was also found to increase with increasing glycerol and protein contents. Overall, results indicated that canola protein isolate films were less brittle, more malleable and transparent, and had greater water vapor permeability at higher glycerol levels. However, as protein level increased, canola protein isolate films were more brittle, less malleable and more opaque, and also had increased water vapor permeability. © The Author(s) 2013 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

  13. Plasma lipid concentrations in preruminant calves fed whole milk with whey protein isolate.

    Science.gov (United States)

    Kushibiki, S; Shingu, H; Komatsu, T; Itoh, F; Hayashi, T; Hodate, K

    2005-02-01

    The objective was to investigate the acute effects of retinol acetate added to whey protein isolate (WPI) on postprandial changes in plasma retinol (experiment 1) and the acute effects of milk fat added to WPI on triglyceride (TG), chylomicrons and very low density lipoprotein (VLDL), and fatty acid concentrations (experiment 2) in suckling calves at 1 and 6 wk of age. In experiment 1, 16 Holstein male calves were alloted to 2 equal groups. On the days of measurement, the calves were fed at 0900 h whole milk [4% of body weight (BW)] mixed with vitamin A acetate (500,000 IU) with or without WPI (0.04% of BW). At 1 wk of age, significantly higher postfeeding concentrations of plasma retinol were observed in the calves fed milk with WPI. At 6 wk of age, no differences in the plasma retinol concentrations were observed between 2 groups. On the days of measurement in experiment 2, 16 male calves were fed at 0900 h whole milk (4% of BW) with added milk fat prepared by centrifugation from whole milk (2% of BW) with or without WPI (0.04% of BW). The milk supplemented with fat was prepared on the day before the measurement. At 1 wk of age, significant higher postfeeding concentrations of plasma TG concentrations were obtained in the calves fed WPI than in the control calves, immediately after the meal or from 7 h later onward. Plasma chylomicrons and VLDL concentrations at 1 wk of age were significantly higher in the WPI-fed group than in the control group at 8 h postfeeding. In the calves with the WPI diet, plasma concentrations of myristic, palmitic, stearic, oleic, and linoleic acids at 1 wk of age were significantly higher than those in the control calves at 8 h after feeding. However, chylomicrons and VLDL, and fatty acid concentrations did not differ between the 2 groups after feeding at 6 wk of age. Results indicate that WPI increases plasma lipid concentration of preruminant calves only at 1 wk of age. These data are interpreted to indicate that WPI enhances mainly

  14. [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.

  15. The influence of whey protein concentrate on growth and survival of probiotic bacteria in whey

    Directory of Open Access Journals (Sweden)

    Ljubica Tratnik

    2008-08-01

    Full Text Available This research examines the influence of whey protein concentrate addition (WPC on growth and activity of probiotic species Lactobacilus acidophilus La-5 and Bifidobacterium animalis subsp. lactis BB-12 in sweet reconstituted whey and their survival during 28 days of fermented whey cold storage (4 °C. The fermentation of whey at 37º C with and without 1.5 and 3% of WPC addition has been observed. Fermentation of whey with Lactobacillus acidophilus La-5 to which 3% WPC was added, was about an hour shorter (∼ 14 hours compared to the fermentation of whey without WPC addition (∼ 15 hours. The viable cells count of Lactobacilus acidophilus La-5 was better in whey with 3% of WPC addition (Δlog CFU/mL = 2.1 compared to whey without WPC addition (Δlog CFU/mL = 1.7 . Addition of whey protein concentrate did not influence significantly on growth of Bifidobacterium animalis subsp. lactis BB-12 in whey, and also it did not influence the survival of both probiotic species during 28 days of cool storage (at 4 °C. Whey enriched with WPC had slightly higher titratable acidity during fermentation and storage compared to whey without WPC addition.

  16. 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

  17. 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

  18. 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

  19. Kinetics of enthalpy relaxation of milk protein concentrate powder upon ageing and its effect on solubility.

    Science.gov (United States)

    Haque, Enamul; Whittaker, Andrew K; Gidley, Michael J; Deeth, Hilton C; Fibrianto, Kiki; Bhandari, Bhesh R

    2012-10-01

    Kinetics of enthalpy relaxation of milk protein concentrate (MPC) powder upon short-term (up to 67 h) storage at 25 °C and aw 0.85, and long-term (up to 48 days) storage at 25 °C and a range of aw values (0-0.85) were studied by differential scanning calorimetry (DSC). The short-term study showed a rapid recovery of enthalpy for the first 48 h, followed by a slower steady increase with time. The non-exponential β parameter was calculated using the Kohlrausch-Williams-Watts function and found to be 0.39. Long-term storage showed that enthalpy relaxation depends on both storage period and water activity. The enthalpy value was much less for lower moisture content (mc) (aw ≤ 0.23, mc ≤ 5.5%) than for higher mc (aw ≥ 0.45, mc ≥ 8%) samples for a particular storage period. The results suggest that the presence of more water molecules, in close proximity to the protein surface facilitates kinetic unfreezing and subsequent motion of molecular segments of protein molecules towards thermodynamic equilibrium. Although de-ageing of stored samples did not reverse storage-induced solubility losses, the timescale of enthalpy relaxation was similar to that of solubility loss. It is suggested that enthalpy relaxation within stored samples allows structural rearrangements that are responsible for subsequent solubility decreases. Copyright © 2012 Elsevier Ltd. All rights reserved.

  20. Effects of enzymatic hydrolysis on the allergenicity of whey protein concentrates.

    Science.gov (United States)

    Duan, Cuicui; Yang, Lijie; Li, Aili; Zhao, Rui; Huo, Guicheng

    2014-08-01

    Cow's milk whey consists of many protein components and some of them are antigens to human and known to modulate immune responses. Enzymatic hydrolysis is a useful method to modify proteins with allergenicity. The objective of this study was to identify whether the in vitro enzymatic hydrolysis could reduce the allergenicity of whey protein concentrates (WPC). In this study, WPC were hydrolyzed by trypsin and twenty-four BALB/c mice were divided into three groups and fed with WPC formula and WPC hydrolysates formula, while the control mice received milk-free diet. The results revealed that there was no significant difference between the body weights among all groups. WPC-fed mice produced an elevated spleen lymphocyte proliferation level than WPC hydrolysates-fed mice and also produced higher levels of WPC-specific IgE in intestinal tract and serum in comparison to WPC hydrolysates-fed mice and control group. Significant up-regulation of plasma histamine levels were also observed and showed the same trend with IgE. The secretions of IL-4 and IL-5 were significantly enhanced by WPC. WPC significantly suppressed the secretion of IFN-γ while hydrolysates of WPC significantly increased the secretion of IFN-γ compared to control group. These results suggest that hydrolysis may play a role to reduce the allergenicity of WPC.

  1. Bacillus thuringiensis Cyt2Aa2 binding on lipid/cholesterol bilayer depends on protein concentration and time.

    Science.gov (United States)

    Tharad, Sudarat; Moreno-Cencerrado, Alberto; Üzülmez, Öykü; Promdonkoy, Boonhiang; Toca-Herrera, Jose L

    2017-10-14

    Bacillus thuringiensis produces cytolytic proteins (Cyt) that show toxicity against dipteran insect larvae acting directly on the cell membrane. Up to now, two different models have been proposed to explain the interaction mechanism of the cytolytic protein Cyt2Aa2 on lipid membranes: pore-forming and detergent-like action. Here we report on the interaction of Cyt2Aa2 with lipid/cholesterol bilayers at early stage (far from equilibrium) as a function of protein concentration. Quartz crystal microbalance with dissipation (QCM-D) measurements showed that the rate of protein adsorption increased with concentration, although the mass of the final protein-lipid was similar after two hours. In addition, the dissipation (compliance of the hybrid lipid/protein layer) increased with decreasing protein concentration. Furthermore, atomic force microscopy (AFM) revealed that the structure of the protein-lipid layer was concentration and time dependent. A rigid hybrid homogeneous layer was observed at protein concentrations of 50 μg/ml and 100 μg/ml after 30 min. At lower concentrations, 10 μg/ml and 17.5 μg/ml, protein adsorption on the lipid layer led to the formation of small aggregates. Interestingly, at 25 μg/ml a transition of a hole-like structure into a homogeneous layer was observed. This suggests that 25 μg/ml is a threshold concentration for the binding mechanism of Cyt2Aa2 on to lipid membranes. Copyright © 2017 Elsevier Inc. All rights reserved.

  2. Chemical composition, molecular weight distribution, secondary structure and effect of NaCl on functional properties of walnut (Juglans regia L) protein isolates and concentrates

    OpenAIRE

    Mao, Xiao-Ying; Hua, Yu-Fei

    2012-01-01

    Chemical composition, molecular weight distribution, secondary structure and effect of sodium chloride concentration on functional properties of walnut protein isolates, concentrates and defatted walnut flour were study. Compared with walnut protein concentrates (75.6%) and defatted walnut flour (52.5%), walnut protein isolates contain a relatively high amount of protein (90.5%). The yield of walnut protein isolates and concentrates was 43.2% and 76.6%, respectively. In molecular weight distr...

  3. 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.

  4. 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.

  5. MODELING OF THE SPRAY DRYING PROCESS OF GREEN PROTEIN SUSPENSION CONCENTRATE (PGC

    Directory of Open Access Journals (Sweden)

    A. A. Shevtsov

    2015-01-01

    Full Text Available Development and implementation of high-tech and energy-efficient methods of feed production is important and ap¬propriate due to the fact that enterprises are not able to provide the market of feed consumers with high quality products at affordable prices. To solve this problem, an alternative technology for the production of protein green concentrate (PGC from the cormophyte mass of high protein plants was developed. The most energy-intensive process of obtaining PGC is spray drying. At the same time the problems of energy saving, and the product quality are solved by modeling. The drying model developed in this study is based on the falling edge of evaporation, which is used in many studies of drops drying. The problem of obtaining the basic equations of heat and mass transfer during the periods of constant and decreasing drying rate was to be solved. It is also supposed that the drying takes place during the periods of constant and decreasing drying rate. Basic equations of heat and mass transfer for both periods of drying were obtained. Changing of thermophysical characteristics were determined by statistical methods in the range of PGC humidity of 10 ... 75% and a temperature of 20 ... 100%. The model is solved by finite difference method with an accuracy of modeling results of 12%. Method of finite differences is a numerical method for solving differential equations based on the replacement of derivative differences schemes and is the grid method. Identification of model parameters to experimental data obtained in the experimental spray dryer was carried out. The solution allows the mathematical model to determine the change in moisture content (DS concentration and drop radial temperature in the spray drying of the PGC concentrate that is necessary both to select the geometrical sizes of the dryer and the drying process parameters controlling.

  6. 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

  7. Review: Amino acid concentration of high protein food products and an overview of the current methods used to determine protein quality.

    Science.gov (United States)

    Huang, Shiqi; Wang, Lydia M; Sivendiran, Thakshi; Bohrer, Benjamin M

    2017-12-04

    Quality of the dietary protein in foods rather than amount of dietary protein may be of greater importance from a human health and wellness standpoint. Various systems are in place to determine the value of dietary protein. Protein digestibility-corrected amino acid score (PDCAAS) and digestible indispensable amino acid score (DIAAS) are the two major protein standards used to determine the completeness of proteins by their unique concentration and digestibility of indispensable amino acids. The purpose of this review was to provide a comprehensive comparison of the amino acid concentration of high protein foods and provide the current status of the use and practicality of the PDCAAS and DIAAS system. This review builds upon previous research analyzing the total nutrient density of protein-rich foods and expands scientific research investigating the quality of proteins. In summary, the average sum of indispensable amino acids for meat and fish products is much more consistent than that of non-meat and plant-based food products. However, some non-meat products have relatively similar amounts of indispensable amino acids on a similar serving size basis. The overwhelming aspect of determining protein quality is that greater research is needed to determine protein digestibility of food products.

  8. 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

  9. 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 ...

  10. 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...

  11. Quantitative proteomics of primary tumors with varying metastatic capabilities using stable isotope-labeled proteins of multiple histogenic origins

    DEFF Research Database (Denmark)

    Lund, Rikke Raaen; Terp, Mikkel Green; Laenkholm, Anne-Vibeke

    2012-01-01

    The development of metastasis is a complex, multistep process that remains poorly defined. To identify proteins involved in the colonization phase of the metastatic process, we compared the proteome of tumors derived from inoculation of a panel of isogenic human cancer cell lines with different...... metastatic capabilities into the mammary fat pad of immunodeficient mice. Using a protein standard generated by SILAC-labeling, a total of 675 proteins was identified and 30 were differentially expressed between at least two of the tumors. The protein standard contained the proteomes of seven cell lines from...

  12. Concentrations of thyroid hormones and iodothyronine binding proteins in serum of the koala (Phascolarctos cinereus).

    Science.gov (United States)

    Lawson, V J; Curtis, A J; Michelangeli, V P; Stockigt, J R; Carrick, F N; Barlow, J W

    1996-07-01

    To examine circulating total and free thyroid hormone (T3 and T4) concentrations, determine serum iodothyronine binding characteristics and estimate thyroid stimulating hormone (TSH) activity in sera of coastal and inland koalas. A prospective study. Koala serum T3 and T4 were measured by radioimmunoassay. T4 binding parameters were determined by radioligand binding and electrophoresis. Koala TSH values were determined by bioassay. Mean total T4 concentrations were 3.2 +/- 2.1 nM although values were significantly higher in inland-dwelling females in comparison to coastal-dwelling males. Free T4 was 3.3 +/- 2.1 pM. Total and free T3 were 0.4 +/- 0.2 nM and 1.4 +/- 0.9 pM respectively, although these values were at the lower end of the assay detection limit and should be viewed with reservation. Electrophoresis of [125I]-T4-labelled serum revealed only two proteins of electrophoretic mobility similar to human transthyretin (TTR) and albumin. Scatchard analysis of T4 binding to serum gave a curvilinear plot, which could be resolved into two binding sites with affinities identical to that of TTR and albumin but both of low concentration. The bioactivity of the TSH present in the sera was measured using a cell line (JP09) transfected with the human TSH receptor. The mean level of stimulation found in the sera corresponded to a bovine TSH activity of less than 10 mU/L. These results suggest that the serum concentrations of free and total thyroid hormones in koalas are low compared to other marsupials and very low compared to eutherian mammals. The mechanism of maintenance of euthyroidism in this species remains to be determined.

  13. Effect of a protein-rich meal on urinary and salivary free amino acid concentrations in human subjects

    NARCIS (Netherlands)

    Brand, H. S.; Jörning, G. G.; Chamuleau, R. A.; Abraham-Inpijn, L.

    1997-01-01

    The aim of the present study was to investigate whether in healthy volunteers acute changes in plasma free amino acid composition after a protein-rich test meal are reflected in the urinary and salivary concentrations of the corresponding amino acids. The ingestion of a protein-rich meal elicited a

  14. C-reactive protein concentration predicts change in body mass index during childhood.

    Directory of Open Access Journals (Sweden)

    Barbara H Lourenço

    Full Text Available OBJECTIVE: Inflammation may constitute an underlying mechanism for increased risk of developing chronic diseases in later years, but few prospective studies have assessed the influence of low-grade inflammation on body weight gain, particularly among children in low- to middle-income settings with lower prevalence of overweight and obesity. We aimed to investigate whether C-reactive protein (CRP, as a biomarker of low-grade inflammation, predicts changes in body mass index-for-age z scores (BAZ during childhood. METHODS: A population-based longitudinal study was conducted in the Brazilian Amazon among children aged ≤10 years in 2007, with follow-up visits in 2009 and 2012. Outcome was annual change in BAZ. As the main exposure of interest, CRP concentrations were divided into four categories, with values 5 years, children in the highest tertile of CRP 5 years.

  15. The effect of diet on protein concentration, hypopharyngeal gland development and virus load in worker honey bees (Apis mellifera L.).

    Science.gov (United States)

    DeGrandi-Hoffman, Gloria; Chen, Yanping; Huang, Eden; Huang, Ming Hua

    2010-09-01

    Elucidating the mechanisms by which honey bees process pollen vs. protein supplements are important in the generation of artificial diets needed to sustain managed honeybees. We measured the effects of diet on protein concentration, hypopharyngeal gland development and virus titers in worker honey bees fed either pollen, a protein supplement (MegaBee), or a protein-free diet of sugar syrup. Workers consumed more pollen than protein supplement, but protein amounts and size of hypopharyngeal gland acini did not differ between the two feeding treatments. Bees fed sugar syrup alone had lower protein concentrations and smaller hypopharyngeal glands compared with the other feeding treatments especially as the bees aged. Deformed wing virus was detected in workers at the start of a trial. The virus concentrations increased as bees aged and were highest in those fed sugar syrup and lowest in bees fed pollen. Overall results suggest a connection between diet, protein levels and immune response and indicate that colony losses might be reduced by alleviating protein stress through supplemental feeding.

  16. 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)

  17. 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.

  18. 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.

  19. 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.

  20. Conducting polymer nanowires for control of local protein concentration in solution

    Science.gov (United States)

    Morris, Joshua D.; Thourson, Scott B.; Panta, Krishna R.; Flanders, Bret N.; Payne, Christine K.

    2017-05-01

    Interfacing devices with cells and tissues requires new nanoscale tools that are both flexible and electrically active. We demonstrate the use of PEDOT:PSS conducting polymer nanowires for the local control of protein concentration in water and biological media. We use fluorescence microscopy to compare the localization of serum albumin in response to electric fields generated by narrow (760 nm) and wide (1.5 µm) nanowires. We show that proteins in deionized water can be manipulated over a surprisingly large micron length scale and that this distance is a function of nanowire diameter. In addition, white noise can be introduced during the electrochemical synthesis of the nanowire to induce branches into the nanowire allowing a single device to control multiple nanowires. An analysis of growth speed and current density suggests that branching is due to the Mullins-Sekerka instability, ultimately controlled by the roughness of the nanowire surface. These small, flexible, conductive, and biologically compatible PEDOT:PSS nanowires provide a new tool for the electrical control of biological systems.

  1. 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.

  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

    2017-12-02

    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. Supplementation with concentrated milk protein in patients undergoing hematopoietic stem cell transplantation.

    Science.gov (United States)

    Perrone, Ana Carolina Amaral de São José; Barbosa, Thaís Rodrigues; da Silva, Fernanda Lopes; Perrone, Ítalo Tuler; de Carvalho, Antônio Fernandes; Stephani, Rodrigo; Dos Santos, Kelli Borges; Atalla, Ângelo; Hallack Neto, Abrahão Elias

    2017-05-01

    The aim of this study was to analyze the influence of dietary supplementation with whey protein concentrate (WPC) in the incidence of oral mucositis (OM) in patients undergoing hematopoietic stem cell transplantation (HSCT). Patients were supplemented with a daily intake of WPC delivering 50% of the daily protein requirements (DPR) according to the Dietary Reference Intakes and classified later based on the amount of ingested supplement until OM median onset. We evaluated 73 patients. Forty-three were part of the historical control and 30 were supplemented with WPC. The OM had a mean duration of 5.3 d (SD 4.5), ranging from the day of the infusion of stem cells until the 17th day after infusion and a median of 5 d after infusion. OM duration was influenced by the conditioning protocol (P WPC (P = 0.01). Patients who consumed the WPC in an amount ≥40% of DPR had a 35% reduction in duration of OM, and the incidence of OM grades 3 and 4 was 11 times smaller. Body mass index, serum albumin, and adverse reactions, such as diarrhea, nausea and vomiting, dysphagia, dry mouth and drooling, showed no statistically significant differences. WPC intake ≥40% of DPR helped to reduce the severity and duration of OM. The use of WPC in patients undergoing HSCT was shown to be safe, encouraging new studies in this population to assess its action mechanism. Copyright © 2016 Elsevier Inc. All rights reserved.

  4. Increased plasma concentrations of vitamin D metabolites and vitamin D binding protein in women using hormonal contraceptives

    DEFF Research Database (Denmark)

    Møller, Ulla K; Streym, Susanna við; Jensen, Lars T

    2013-01-01

    UNLABELLED: Use of hormonal contraceptives (HC) may influence total plasma concentrations of vitamin D metabolites. A likely cause is an increased synthesis of vitamin D binding protein (VDBP). Discrepant results are reported on whether the use of HC affects free concentrations of vitamin D...

  5. Surfactant protein B and A concentrations are increased in neonatal pneumonia.

    Science.gov (United States)

    D'Aronco, Sara; Simonato, Manuela; Vedovelli, Luca; Baritussio, Aldo; Verlato, Giovanna; Nobile, Stefano; Giorgetti, Chiara; Nespeca, Matteo; Carnielli, Virgilio P; Cogo, Paola E

    2015-10-01

    Term newborns with pneumonia show a reduced pulmonary compliance due to multiple and ill-defined factors. Surfactant proteins' (SPs) changes could have a role in the reduced compliance but the matter is still unsettled. The aim of this study was to clarify the meaning of SPs changes during pneumonia in term newborns. In 28 term ventilated newborns, 13 with pneumonia and 15 with no lung disease, we measured SP-B, SP-A, disaturated-phosphatidylcholine (DSPC), and total phospholipids (PL) concentrations in tracheal aspirates at intubation and close to extubation. We also measured DSPC kinetics using (U-(13)C-PA)dipalmitoyl-phosphatidylcholine. At baseline, SP-B, expressed as % of PL, was significantly different between the groups, being 3.5-fold higher in pneumonia than controls. Conversely, SP-A did not vary between the groups. At extubation, SP-B and SP-A concentrations had decreased significantly in newborns with pneumonia, while there was no significant change in controls. DSPC t1/2 was significantly shorter in the pneumonia group (11.8 (5.5-19.8) h vs. 26.6 (19.3-63.6) h, P = 0.011). In term newborns with pneumonia, SP-B increases with respect to PL, and DSPC is turned over at a faster rate. Disease's resolution is associated with the restoration of the normal ratio between SP-B and PL.

  6. 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

  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. Comparative evaluation of postmortem serum concentrations of neopterin and C-reactive protein.

    Science.gov (United States)

    Ishikawa, Takaki; Hamel, Marianne; Zhu, Bao-Li; Li, Dong-Ri; Zhao, Dong; Michiue, Tomomi; Maeda, Hitoshi

    2008-08-06

    The cellular immune response is accompanied by the release of neopterin. The level of neopterin in serum is increased in patients suffering from viral infections, autoimmune diseases, systemic inflammation, allograft rejection and malignant diseases, while that of C-reactive protein (CRP) is known to rise during inflammatory diseases and traumas. To investigate postmortem neopterin and CRP concentrations with regard to the cause of death, we examined cardiac and peripheral blood samples in 474 autopsy cases without advanced decomposition (0-96 years of age, 343 males and 131 females), 2.8 h to 3 days (median, 18.0 h) after death. Survival time was 0.1 h to 5 months (median, 3.0 h) for traumatic death, and 0.1-1, 440 h (median, 2.5 h) for natural death. In autopsied subjects, neopterin concentrations were higher than the clinical reference, independent of the time after death, and depended on the survival time. In cases of acute and subacute death due to trauma, the neopterin level in right heart blood was mildly to moderately elevated (about 50-200 nmol/l) except for sharp instrument injury, whereas the CRP concentration usually remained low (in the CRP level (around 1-10 mg/dl) was observed in fatal cases of hypothermia (cold exposure). Markedly elevated serum CRP and neopterin levels (>10 mg/dl and >500 nmol/l, respectively) were detected in cases of delayed death due to trauma involving systemic inflammatory response syndrome (SIRS) and of fatal bacterial infections. For sepsis, the serum CRP level was markedly elevated but the neopterin level was low in some cases. Fatal viral infections usually resulted in a marked elevation in the serum neopterin level (>500 nmol/l) with a mild to moderate rise in the CRP level. Combined analyses of neopterin and CRP may be useful to investigate viral infections and delayed traumatic death involving SIRS to support pathological findings.

  9. 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...

  10. 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

  11. Influence of pH and salt concentration on functional properties of walnut protein from different extraction methods.

    Science.gov (United States)

    Hu, Haifang; Fan, Tao; Zhao, Xiaoyan; Zhang, Xiaowei; Sun, Yali; Liu, Hongkai

    2017-08-01

    The effect of pH and NaCl on solubility and functional properties of walnut proteins obtained through AOT reverse micelles, enzyme-assisted reverse micelles and aqueous phase extraction methods was investigated and compared. Extraction yield, foaming properties, water holding capacities of protein obtained through enzyme-assisted reverse micelles at pH 2-12 and NaCl concentration 0.1-1 M were significantly higher than those of the AOT reverse micelles and aqueous phase extracted two. The solubility of proteins by AOT reverse micelles and enzyme-assisted reverse micelles at certain pH and NaCl concentration had no significant difference, but was higher that of the aqueous buffer. Oil holding capacity of three proteins was 2.35, 3.96 and 1.08 cm 3 /g, respectively. At pH 6-12 and NaCl concentration 0.1-1 M, the emulsifying activity of protein from AOT reverse micelles was higher than those of other two methods, while the emulsifying stability of protein from enzyme-assisted reverse micelles was the highest. The functional properties of walnut proteins were affected by extraction methods. It indicated that the walnut protein might be potentially applied in food industry as a food ingredient.

  12. Fusion of the Dhfr/Mtx and IR/MAR gene amplification methods produces a rapid and efficient method for stable recombinant protein production.

    Science.gov (United States)

    Noguchi, Chiemi; Araki, Yoshio; Miki, Daisuke; Shimizu, Noriaki

    2012-01-01

    Amplification of the dihydrofolate reductase gene (Dhfr) by methotrexate (Mtx) exposure is commonly used for recombinant protein expression in Chinese hamster ovary (CHO) cells. However, this method is both time- and labor-intensive, and the high-producing cells that are generated are frequently unstable in culture. Another gene amplification method is based on using a plasmid bearing a mammalian replication initiation region (IR) and a matrix attachment region (MAR), which result in the spontaneous initiation of gene amplification in transfected cells. The IR/MAR and Dhfr/Mtx methods of gene amplification are based on entirely different principles. In this study, we combine these two methods to yield a novel method, termed the IR/MAR-Dhfr fusion method, which was used to express three proteins, the Fc receptor, GFP, and recombinant antibody. The fusion method resulted in a dramatic increase in expression of all three proteins in two CHO sub-lines, DXB-11, and DG44. The IR/MAR-Dhfr fusion amplified the genes rapidly and efficiently, and produced larger amounts of antibody than the Dhfr/Mtx or IR/MAR methods alone. While the amplified structure produced by the Dhfr/Mtx method was highly unstable, and the antibody production rate rapidly decreased with the culture time of the cells, the IR/MAR-Dhfr fusion method resulted in stable amplification and generated clonal cells that produced large amounts of antibody protein over a long period of time. In summary, the novel IR/MAR-Dhfr fusion method enables isolation of stable cells that produce larger amounts of a target recombinant protein more rapidly and easily than either the Dhfr/Mtx or IR/MAR methods alone.

  13. 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

    for these phenomena can be due to short range electrostatic and/or hydrophobic protein-protein interactions. Hydrogen/deuterium exchange mass spectrometry (HDX-MS) is a useful tool for investigating protein conformation, dynamics, and interactions. However, "traditional" continuous dilution labeling HDX......-MS experiments have limited utility for the direct analysis of solutions with high concentrations of protein. Here, we present a dialysis-based HDX-MS (di-HDX-MS) method as an alternative HDX-MS labeling format, which takes advantage of passive dialysis rather than the classic dilution workflow. We applied...

  14. 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

  15. Stable isotope labeling by amino acids in cell culture based proteomics reveals differences in protein abundances between spiral and coccoid forms of the gastric pathogen Helicobacter pylori.

    Science.gov (United States)

    Müller, Stephan A; Pernitzsch, Sandy R; Haange, Sven-Bastiaan; Uetz, Peter; von Bergen, Martin; Sharma, Cynthia M; Kalkhof, Stefan

    2015-08-03

    Helicobacter pylori (H. pylori) is a ε-proteobacterium that colonizes the stomach of about half of the world's population. Persistent infections have been associated with several gastric diseases. Mainly rod- or spiral shaped but also coccoid H. pylori forms have been isolated from mucus layer biopsies of patients. It is still being debated whether the coccoid form can be transformed back into the spiral form or whether this morphology is a result of bacterial cell death or persistence. We established stable isotope labeling by amino acids in cell culture (SILAC) for quantitative proteomics of H. pylori and applied it to investigate differences between the spiral and the coccoid morphology. We detected 72% and were able to relatively quantify 47% of the H. pylori proteome. Proteins involved in cell division and transcriptional and translational processes showed a lower abundance in coccoid cells. Additionally, proteins related to host colonization, including CagA, the arginase RocF, and the TNF-α inducing protein were down-regulated. The fact that outer membrane proteins were observed at higher abundances might represent a mechanism for immune evasion but also preserves adherence to host cells. The established protocol for relative protein quantification of H. pylori samples offers new possibilities for research on H. pylori. Our study shows that SILAC can be employed to study protein abundance changes in H. pylori. We have chosen to establish SILAC for H. pylori because it facilitates fractionation on both, protein and peptide level and thus enables deep proteome coverage. Furthermore, SILAC allows robust and highly accurate protein quantification. The manuscript includes a detailed description of the applied method, suggestions for further improvement as well as a practical application. The investigation of differences between the coccoid and infectious spiral morphology of H. pylori with SILAC revealed the regulation of proteins that are involved in host

  16. Conformational Analysis of Proteins in Highly Concentrated Solutions by Dialysis-Coupled Hydrogen/Deuterium Exchange Mass Spectrometry

    Science.gov (United States)

    Houde, Damian; Nazari, Zeinab E.; Bou-Assaf, George M.; Weiskopf, Andrew S.; Rand, Kasper D.

    2016-04-01

    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 for these phenomena can be due to short range electrostatic and/or hydrophobic protein-protein interactions. Hydrogen/deuterium exchange mass spectrometry (HDX-MS) is a useful tool for investigating protein conformation, dynamics, and interactions. However, "traditional" continuous dilution labeling HDX-MS experiments have limited utility for the direct analysis of solutions with high concentrations of protein. Here, we present a dialysis-based HDX-MS (di-HDX-MS) method as an alternative HDX-MS labeling format, which takes advantage of passive dialysis rather than the classic dilution workflow. We applied this approach to a highly concentrated antibody solution without dilution or significant sample manipulation, prior to analysis. Such a method could pave the way for a deeper understanding of the unusual behavior of proteins at high concentrations, which is highly relevant for development of biopharmaceuticals in industry.

  17. 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

  18. Application of a PEG precipitation method for solubility screening: A tool for developing high protein concentration formulations

    Science.gov (United States)

    Li, Li; Kantor, Angela; Warne, Nicholas

    2013-01-01

    Previous publications demonstrated that the extrapolated solubility by polyethylene glycol (PEG) precipitation method (Middaugh et al., J Biol Chem 1979; 254:367–370; Juckes, Biochim Biophys Acta 1971; 229:535–546; Foster et al., Biochim Biophys Acta 1973; 317:505; Mahadevan and Hall, AIChE J 1990; 36:1517–1528; Stevenson and Hageman, Pharm Res 1995; 12:1671–1676) has a strong correlation to experimentally measured solubility of proteins. Here, we explored the utility of extrapolated solubility as a method to compare multiple protein drug candidates when nonideality of a highly soluble protein prohibits accurate quantitative solubility prediction. To achieve high efficiency and reduce the amount of protein required, the method is miniaturized to microwell plate format for high-throughput screening application. In this simplified version of the method, comparative solubility of proteins can be obtained without the need of concentration measurement of the supernatant following the precipitation step in the conventional method. The monoclonal antibodies with the lowest apparent solubilities determined by this method are the most difficult to be concentrated, indicating a good correlation between the prediction and empirical observations. This study also shows that the PEG precipitation method gives results for opalescence prediction that favorably compares to experimentally determined opalescence levels at high concentration. This approach may be useful in detecting proteins with potential solubility and opalescence problems prior to the time-consuming and expensive development process of high concentration formulation. PMID:23740802

  19. Protein Immobilization Capabilities of Sucrose and Trehalose Glasses: The Effect of Protein/Sugar Concentration Unraveled by High-Field EPR.

    Science.gov (United States)

    Malferrari, Marco; Savitsky, Anton; Lubitz, Wolfgang; Möbius, Klaus; Venturoli, Giovanni

    2016-12-01

    Disaccharide glasses are increasingly used to immobilize proteins at room temperature for structural/functional studies and long-term preservation. To unravel the molecular basis of protein immobilization, we studied the effect of sugar/protein concentration ratios in trehalose or sucrose matrixes, in which the bacterial photosynthetic reaction center (RC) was embedded as a model protein. The structural, dynamical, and H-bonding characteristics of the sugar-protein systems were probed by high-field W-band EPR of a matrix-dissolved nitroxide radical. We discovered that RC immobilization and thermal stabilization, being independent of the protein concentration in trehalose, occur in sucrose only at sufficiently low sugar/protein ratios. EPR reveals that only under such conditions does sucrose form a microscopically homogeneous matrix that immobilizes, via H-bonds, the nitroxide probe. We conclude that the protein immobilization capability depends critically on the propensity of the glass-forming sugar to create intermolecular H-bond networks, thus establishing long-range, homogeneous connectivity within the matrix.

  20. 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.

  1. 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.

  2. The effect of bleaching agent on the flavor of liquid whey and whey protein concentrate.

    Science.gov (United States)

    Croissant, A E; Kang, E J; Campbell, R E; Bastian, E; Drake, M A

    2009-12-01

    The increasing use and demand for whey protein as an ingredient requires a bland-tasting, neutral-colored final product. The bleaching of colored Cheddar whey is necessary to achieve this goal. Currently, hydrogen peroxide (HP) and benzoyl peroxide (BPO) are utilized for bleaching liquid whey before spray drying. There is no current information on the effect of the bleaching process on the flavor of spray-dried whey protein concentrate (WPC). The objective of this study was to characterize the effect of bleaching on the flavor of liquid and spray-dried Cheddar whey. Cheddar cheeses colored with water-soluble annatto were manufactured in duplicate. Four bleaching treatments (HP, 250 and 500 mg/kg and BPO, 10 and 20 mg/kg) were applied to liquid whey for 1.5 h at 60 degrees C followed by cooling to 5 degrees C. A control whey with no bleach was also evaluated. Flavor of the liquid wheys was evaluated by sensory and instrumental volatile analysis. One HP treatment and one BPO treatment were subsequently selected and incorporated into liquid whey along with an unbleached control that was processed into spray-dried WPC. These trials were conducted in triplicate. The WPC were evaluated by sensory and instrumental analyses as well as color and proximate analyses. The HP-bleached liquid whey and WPC contained higher concentrations of oxidation reaction products, including the compounds heptanal, hexanal, octanal, and nonanal, compared with unbleached or BPO-bleached liquid whey or WPC. The HP products were higher in overall oxidation products compared with BPO samples. The HP liquid whey and WPC were higher in fatty and cardboard flavors compared with the control or BPO samples. Hunter CIE Lab color values (L*, a*, b*) of WPC powders were distinct on all 3 color scale parameters, with HP-bleached WPC having the highest L* values. Hydrogen peroxide resulted in a whiter WPC and higher off-flavor intensities; however, there was no difference in norbixin recovery between HP

  3. Isolation of recombinant proteins from culture broth by co-precipitation with an amino acid carrier to form stable dry powders.

    Science.gov (United States)

    Moore, Barry D; Deere, Joseph; Edrada-Ebel, RuAngelie; Ingram, Andrew; van der Walle, Christopher F

    2010-08-01

    Protein-coated microcrystals can be generated by co-precipitation of protein and a water-soluble crystalline carrier by addition to excess water miscible organic solvent. We have investigated this novel process for its utility in the concentration and partial purification of a recombinant protein exported into the culture broth during expression by Pichia pastoris. Co-precipitation with a L-glutamine carrier selectively isolated the protein content of the culture broth, with a minimal number of steps, and simultaneously removed contaminants including a novel yeast metabolite. This pigment co-elutes during aqueous chromatography but its elucidation as a benzoylated glycosamine suggested a simple route of removal by partition during the co-precipitation process. Scale-up of the process was readily achieved through in-line mixing and subsequent reconstitution of the dried protein-coated microcrystals yielded natively folded, bioactive protein. Additional washing of the crystals with saturated L-glutamine facilitated further purification of the recombinant protein immobilized on the L-glutamine carrier. Thus, we present a novel method for the harvesting of recombinant protein from culture broth as a dry powder, which may be of general applicability to bioprocessing. (c) 2010 Wiley Periodicals, Inc.

  4. Nitrate concentration-shift cultivation to enhance protein content of heterotrophic microalga Chlorella vulgaris: Over-compensation strategy.

    Science.gov (United States)

    Xie, Tonghui; Xia, Yun; Zeng, Yu; Li, Xingrui; Zhang, Yongkui

    2017-06-01

    Protein production from microalgae requires both high cell density during cultivation and high protein content in cells. Heterotrophic microalgae can achieve high cell density, and yet are confronted with the problem of low protein content. Based on over-compensation strategy, a new concentration-shift method was proposed to cultivate heterotrophic Chlorella vulgaris, aiming to increase protein content. With a prior starvation period, microalgae utilized more nitrate and accumulated more proteins compared to one-stage cultivation. Considering the convenience of operation, nitrate-added culture was adopted for producing heterotrophic microalgae, rather than sterile centrifugal culture. Operating parameters including nitrate concentration in N-deficient medium, N-starved time and nitrate concentration in N-rich medium were optimized, which were 0.18gl -1 , 38h and 2.45gl -1 , respectively. Under the optimized conditions, protein content in heterotrophic Chlorella reached 44.3%. Furthermore, the heterotrophic microalga was suggested to be a potential single-cell protein source according to the amino acid composition. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Association of Plasma Heat Shock Protein 70, Interleukin 6, and Creatine Kinase Concentrations in a Healthy, Young Adult Population

    Directory of Open Access Journals (Sweden)

    Carmen Contreras-Sesvold

    2015-01-01

    Full Text Available Variations of baseline plasma concentrations of creatine kinase (CK, heat shock protein 70 (HSP70, and interleukin 6 (IL-6 have been reported. We report categorical associations which may influence these protein levels. Methods. Blood was harvested for DNA and plasma protein analysis from 567 adults. Mean protein levels of CK, HSP70, and IL-6 were compared by sex, ethnicity, genetic variants—CKMM Nco1 (rs1803285, HSPA1B +A1538G (rs1061581, and IL6 G-174C (rs1800795—self-reported history of exercise, oral contraceptive use, and dietary supplement use. Results. SNP major allele frequencies for CKMM, HSPA1B, and IL6 were 70% A, 57% A, and 60%. Mean CK statistically differed by sex, ethnicity, oral contraceptives, and caffeine. Plasma HSP70 differed by caffeine and protein. Mean IL-6 concentration differed by sex, ethnicity, and genotype. Plasma IL-6 was significantly lower (29% in males (1.92 ± 0.08 pg/mL and higher (29% among African Americans (2.85 ± 0.50 pg/mL relative to the others. IL6 G-174C GG genotype (2.23 ± 0.14 pg/mL was 19% greater than CG or CC genotypes. Conclusion. Differences in baseline CK and IL-6 plasma protein concentrations are associated with genetics, sex, ethnicity, and the use of oral contraceptives, caffeine, and protein supplements in this young and athletic population.

  6. Sensory aroma characteristics of alcalase hydrolyzed rice bran protein concentrate as affected by spray drying and sugar addition.

    Science.gov (United States)

    Arsa, Supeeraya; Theerakulkait, Chockchai

    2015-08-01

    The sensory aroma characteristics of alcalase hydrolyzed rice bran protein concentrate as affected by spray drying and sugar addition were investigated. Rice bran protein concentrate (RBPC) was hydrolyzed by alcalase. Sucrose, glucose or fructose was added to the liquid rice bran protein hydrolysate (LRBPH) and subsequently spray dried. The sensory aroma intensities of the hydrolysates were evaluated. Results showed that after spray drying, the rice bran protein concentrate powder (RBPC-P) had higher sweet and cocoa-like aroma intensities than RBPC (p ≤ 0.05) and hydrolyzed rice bran protein powder (HRBPP) had higher milk powder-like aroma intensities than LRBPH (p ≤ 0.05). The sweet, cocoa-like and milk powder-like aroma intensities in hydrolyzed rice bran protein powder with fructose addition (HRBPP-F) were significantly higher (p ≤ 0.05) than those of hydrolyzed rice bran protein powder with sucrose or glucose addition (HRBPP-S or HRBPP-G). HRBPP-F had the highest overall aroma liking score. These results also indicate that spray drying and sugar addition could improve the sensory aroma characteristics of alcalase hydrolyzed RBPC.

  7. Serum protein and casein concentration: effect on pH and freezing point of milk with added CO2.

    Science.gov (United States)

    Ma, Y; Barbano, D M

    2003-05-01

    The objective of this study was to determine the effect of protein concentration and protein type [i.e., casein (CN) and serum protein (SP)] on pH (0 degree C) and freezing point (FP) of skim milk upon CO2 injection at 0 degree C. CN-free skim milks with increasing SP content (0, 3, and 6%) and skim milks with the same SP content (0.6%) but increasing CN content (2.4, 4.8, and 7.2%) were prepared using a combination of microfiltration and ultrafiltration processes. CO2 was injected into milks at 0 degree C using a continuous flow carbonation unit (230 ml/min). Increasing SP or CN increased milk buffering capacity and protein-bound mineral content. At the same CO2 concentration at 0 degree C, a milk with a higher SP or a higher CN concentration had more resistance to pH change and a greater extent of FP decrease. The buffering capacity provided by an increase of CN was contributed by both the CN itself and the colloidal salts solublized into the serum phase from CN upon carbonation. Skim milks with the same true protein content (3%), one with 2.4% CN plus 0.6% SP and one with 3% SP, were compared. At the same true protein content (3%), increasing the proportion of CN increased milk buffering capacity and protein-bound mineral content. Milk with a higher proportion of CN had more resistance to pH change and a greater extent of FP decrease at the same carbonation level at 0 degree C. Once CO2 was dissolved in the skim portion of a milk, the extent of pH reduction and FP depression depended on protein concentration and protein type (i.e., CN and SP).

  8. Stable expression of foot-and-mouth disease virus protein VP1 fused with cholera toxin B subunit in the potato (Solanum tuberosum).

    Science.gov (United States)

    He, Dong-Mei; Qian, Kai-Xian; Shen, Gui-Fang; Li, Yi-Nü; Zhang, Zhi-Fang; Su, Zhong-Liang; Shao, Hong-Bo

    2007-04-01

    The expression vector, pBI121CTBVP1, containing the fusion of the foot and mouth disease virus (FMDV) VP1 gene and the cholera toxin B subunit (CTB) gene was constructed by fused PCR and transferred into potato (Solanum tuberosum L.) by Agrobacterium-mediated transformation. Transformed plants were obtained by selecting on kanamycin-resistant medium strictly and regenerated. The transgenic plantlets were identified by PCR, Southern-blot and the production of fused protein was confirmed and quantified by Western-blot and ELISA assays. The results showed that the fused genes were expressed stablely under the control of specific-tuber patatin promoter. The expressed fused proteins have a certain degree of immunogenicity.

  9. Fluctuations in C-reactive protein concentration and neutrophil activation during normal human pregnancy.

    Science.gov (United States)

    Belo, Luís; Santos-Silva, Alice; Rocha, Susana; Caslake, Muriel; Cooney, Josephine; Pereira-Leite, Luís; Quintanilha, Alexandre; Rebelo, Irene

    2005-11-01

    To clarify the changes in serum C-reactive protein (CRP) levels and in the neutrophil activation state during normal human pregnancy. A longitudinal study (n=23) was performed during the three trimesters of pregnancy; a group of non-pregnant women (n=24) was used as control. Total and differential leukocyte count, serum concentration of CRP and plasma levels of granulocyte-macrophage colony stimulating factor (GM-CSF) and of lactoferrin and elastase (two indirect markers of neutrophil activation) were measured. Pregnancy imposed an inflammatory response in the mother, observed by the significant increment in total white blood cell (WBC) and neutrophil counts and in the circulating levels of CRP, GM-CSF and lactoferrin, in all trimesters of gestation compared with non-pregnant controls. Plasma elastase concentration was also significantly higher in pregnant women, but only in the first trimester of gestation. Regarding the ratios of lactoferrin and elastase per neutrophil, they were significantly lower in pregnant women (all trimesters). During gestation, WBC and neutrophil count increased significantly from the first to the second trimester and remained high in the third period. In contrast, the ratios of lactoferrin and elastase per neutrophil decreased significantly from the first to the second trimester, remaining low in the last trimester. Concerning CRP levels, no consistent changes were observed throughout gestation; 12 cases (52.2%) presented fluctuations, whereas 7 (30.4%) showed progressive reductions and 4 (17.4%) progressive increments throughout pregnancy. Changes in CRP levels vary in a wide manner between subjects along pregnancy, even though median values are consistently elevated throughout pregnancy. Moreover, circulating levels of neutrophil-activation products are higher in normal human gestation.

  10. Influence of alfalfa protein concentrate dietary supplementation on blood parameters of growing-finishing pigs

    Directory of Open Access Journals (Sweden)

    Pietrzak Krzysztof

    2015-09-01

    Full Text Available The aim of the study was to analyse the effect of the feed additive alfalfa protein concentrate (APC, on pig health. The trial involved 40 crossbred gilts and 40 crossbred castrates (Polish Landrace × Polish Large White × Duroc of 29.0 ± 0.5 kg initial body weight. Allocation of experimental animals was into four treatment groups: the control group (C was fed standard mixtures, without APC addition; group E-15 was fed a basal diet supplemented with 1.5% APC; and groups E-30 and E-30P were fed diets with 3.0% APC inclusion. There were two feeding systems. In the first system, animals of groups C, E-15, and E-30 were fed continuously with suitable mixtures. The second feeding system was used in group E-30P where animals received the experimental or control mixture alternating at two-week intervals. The addition of APC supplement to diets significantly increased (P ≤ 0.05 red blood cell indices, i.e. haematocrit (Ht, red blood cell count (RBC, and haemoglobin concentration (Hb in growing and finishing periods. The analysis of enzyme activity demonstrated a markedly higher activity of aspartate aminotransferase (AST, alanine aminotransferase (ALT, and especially alkaline phosphatase (ALP in the blood plasma of pigs fed APC supplement. This increase may indicate a negative impact of APC on the animal’s liver. A positive effect of dietary APC on blood lipid parameters was associated with a decreased level of total cholesterol and reduced low-density lipoprotein fraction. Analysis of the haematological and biochemical blood indices demonstrated that APC additive may affect animal health.

  11. 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

  12. 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.

  13. 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.

  14. Growth of ABT-5 bacteria culture in whey supplemented with whey protein concentrate

    Directory of Open Access Journals (Sweden)

    Bojan Matijević

    2010-09-01

    Full Text Available The objective of this research was to decrease the duration of whey fermentation and to analyze the impact of whey protein concentrates (WPC addition on growth and activity as well as on survival of mixed ABT-5 culture bacteria (Lactobacillus acidophilus, Bifidobacterium spp. and Streptococcus thermophilus during 28 days of fermented whey cold storage (5 °C. Whey fermentation was observed at 37 °C with and without 1.5 and 3 % WPC addition. Whey fermentation with mixed ABT-5 bacterial culture took about 6.5 hours, and WPC did not affect the fermentation dynamics, regardless of the added amount. The viable cell count of Lactobacillus acidophilus was higher in fermented whey with WPC (about 7.13 log CFU/mL compared to whey without WPC (about 6.63 log CFU/mL. WPC addition did not affect the growth of bacteria Bifidobacterium spp. and Streptococcus thermophilus nor the survival of ABT-5 culture bacteria in fermented whey during 28 days of cold storage (5 °C.

  15. Hydrophobicity, thermal and micro-structural properties of whey protein concentrate-pullulan-beeswax films.

    Science.gov (United States)

    Jafari, Seid Mahdi; Khanzadi, Mehrdad; Mirzaei, Habibollah; Dehnad, Danial; Chegini, Faramarz Khodaian; Maghsoudlou, Yayha

    2015-09-01

    In this research, effects of beeswax (BW) on functional properties of whey protein concentrates (WPC):pullulan (PUL) films were investigated. For this purpose, 0, 10, 20 and 30w/w(glycerol)% BW rates and 30:70, 50:50 and 70:30w/w% WPC:PUL ratios were applied. Films containing 70% WPC:30% PUL (WPC70) and 30% BW (BW30) justified the highest contact angle (92.4°) among all films; SEM micrographs indicated that BW could come toward the surface of films during drying stage and resulted in a higher hydrophobic behavior of bilayer films compared with blend films. WPC70 supplied the lowest T(g) values (36-48 °C) among different proportions of WPC-PUL; the highest melting points were just assured in the absence of BW regardless of combination ratio for WPI:PUL. BW30 films deserved lower roughness rates than BW20 (and even BW10) films, indicating more advantageous microstructure and higher hydrogen connections in BW30 films and justifying similar melting points attained for BW30 films to BW20 or 10 ones. Overall, application of WPC70 and BW30 was recommended to obtain optimum combination of final properties for WPC-PUL-BW bilayer films as SEM exhibited flexible and elastic structures of such films. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. 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.

  17. 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.

  18. Effect of protein concentrates, emulsifiers on textural and sensory characteristics of gluten free cookies and its immunochemical validation.

    Science.gov (United States)

    Sarabhai, Swati; Indrani, D; Vijaykrishnaraj, M; Milind; Arun Kumar, V; Prabhasankar, P

    2015-06-01

    The effect of 5, 7.5 and 10 % protein concentrates namely soya protein isolate (SPI), whey protein concentrate (WPC) and addition of 0.5 % emulsifiers such as glycerol monostearate (GMS), sodium stearoyl- 2- lactylate (SSL) and lecithin (LEC) on the rheological, sensory and textural characteristics of cookies with rice flour and its immunochemical validation was studied. The results showed that the use of 7.5 % SPI/WPC along with GMS significantly improved the quality characteristics of cookies with rice flour. Dot-Blot and Western-blot studies of cookies with 7.5 % of SPI or WPC confirmed that the anti-gliadin did not recognize these proteins. Carry- through process using ELISA kit confirmed that gluten was within the permissible limit in all the stages of processing and hence these cookies can be consumed by people suffering from celiac disease.

  19. 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)

  20. The Achilles' Heel of "Ultrastable" Hyperthermophile Proteins: Submillimolar Concentrations of SDS Stimulate Rapid Conformational Change, Aggregation, and Amyloid Formation in Proteins Carrying Overall Positive Charge.

    Science.gov (United States)

    Khan, Javed M; Sharma, Prerna; Arora, Kanika; Kishor, Nitin; Kaila, Pallavi; Guptasarma, Purnananda

    2016-07-19

    Low concentrations (SDS) have been shown to induce the formation of amyloid fibers in more than 20 different mesophile-derived proteins in the cationic state. It is not known whether SDS has similar effects on hyperthermophile-derived proteins, which are otherwise thought to be "ultrastable" and inordinately resistant to structural perturbations at room temperature. Here, we show that low (SDS rapidly induce the formation of aggregates and amyloid fibers in five different ultrastable Pyrococcus furiosus proteins in the cationic state. We also show that amyloid formation is accompanied by the development of a characteristic, negative circular dichroism band at ∼230 nm. These effects are not seen if the proteins have a net negative charge or when higher concentrations of SDS are used (which induce helix formation instead). Our results appear to reveal a potential weakness or "Achilles' heel" in ultrastable proteins from hyperthermophiles. They also provide very strong support for the view that SDS initially interacts with proteins through electrostatic interactions, and not hydrophobic interactions, eliciting similar effects entirely regardless of protein molecular weight, or structural features such as quaternary structure or tertiary structural stability.

  1. 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...... protein under future climate change conditions....

  2. The Prognostic Significance of the Serum p53 Protein Concentration in Chinese Patients with Non-Hodgkin Lymphoma

    Directory of Open Access Journals (Sweden)

    Min Zhou

    2012-12-01

    Full Text Available OBJECTIVE: To investigate the prognostic significance of cytogenetic abnormalities, staging, patient factors, and the serum p53 protein concentration in Chinese non-Hodgkin lymphoma (NHL patients. METHODS: The study included 43 patients with NHL that were identified between August 2003 and December 2008. Patient clinical characteristics patients were determined based on morphological, immunohistochemical, and cytogenetic analysis, and the serum p53 protein concentration was measured quantitatively. RESULTS: Following conventional chemotherapy, the complete/partial remission (CR/PR rate was significantly higher and overall survival (OS was significantly longer in the patients with early-stage (stage I-II lymphoma, normal karyotype, and a low serum p53 protein concentration than in those with advanced-stage (stage III-IV lymphoma, cytogenetic abnormalities, and a high serum p53 protein concentration (≥0.35 U/mL. Bone marrow infiltration was also a predictor of poor response and OS. There weren’t any significant differences in disease remission between the male and female patients, older and younger patients (aged <70 years vs. ≥70 years, or B-cell lymphoma and T-cell lymphoma patients. CONCLUSION: Staging is an effective means of assessing the severity of NHL. Cytogenetic examination can provide useful information for diagnosis, staging, and prognostication. The serum p53 protein level may be a potential prognostic marker in patients with NHL.

  3. 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

  4. Functional Aspects of the Solution Structure and Dynamics of PAF, a Highly Stable Antifungal Protein from Penicillium chrysogenum

    Science.gov (United States)

    Batta, Gyula; Barna, Teréz; Gáspári, Zoltán; Sándor, Szabolcs; Kövér, Katalin E.; Binder, Ulrike; Sarg, Bettina; Kaiserer, Lydia; Chhillar, Anil Kumar; Eigentler, Andrea; Leiter, Éva; Hegedüs, Nikoletta; Pócsi, István; Lindner, Herbert; Marx, Florentine

    2015-01-01

    Summary PAF is a promising antimycotic without toxic effects on mammalian cells and therefore may be a drug candidate against the often lethal Aspergillus infections in human. The pathogenesis of PAF on sensitive fungi involves G-protein coupled signaling followed by apoptosis. Here, the solution structure of this small, cationic, antifungal protein from Penicillium chrysogenum is determined by solution NMR. We proved that PAF belongs to the SCOP fold class of its closest homologue AFP from Aspergillus giganteus. PAF comprises five β—strands forming two orthogonally packed β—sheets sharing a common interface. The ambiguity in the assignment of two disulfide bonds of three was investigated by NMR dynamics combined with restrained molecular dynamics calculations. The clue could not be resolved: two ensembles with different disulfide patterns and the one with no S-S bond exhibit essentially the same fold. 15N relaxation dispersion and interference experiments did not reveal disulfide bond rearrangements via slow exchange. The measured order parameters and the 3.0 ns correlation time is appropriate for a compact monomeric protein of this size. We demonstrated by site-directed mutagenesis that the highly conserved and positively charged lysine-rich domain region on the surface enhances the toxicity of PAF. However, the efficacy of the OB fold is reduced in PAF compared to AFP, due to less solvent exposed aromatic regions explaining the absence of chitobiose binding. The present work lends further support to the understanding of the documented substantial differences between the mode of action of two highly homologous antifungal proteins. PMID:19459942

  5. 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)

  6. 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

  7. Effects of moisture and temperature during grain storage on the functional properties and isoflavone profile of soy protein concentrate.

    Science.gov (United States)

    Ziegler, Valmor; Ferreira, Cristiano Dietrich; Hoffmann, Jéssica Fernanda; de Oliveira, Maurício; Elias, Moacir Cardoso

    2018-03-01

    The objective of this study was evaluate the effects of moisture and temperature during grain storage on the functional properties and isoflavone profile of soy protein concentrate. The protein concentrate was extracted from soybeans on the first day of storage and after 12months of storage, at 12 and 15% moisture and at temperatures of 11, 18, 25, and 32°C. The protein concentrate obtained from grains stored at 32°C showed decreased extraction yield (51.6% with 15% of moisture) and protein solubility (24.8% with 15% of moisture) with increased emulsifying capacity (52.2% with 12% of moisture) and foam formation, compared with sample obtained at the start of the storage period. The isoflavone profile showed a decrease in the concentrations of glycosylated, malonyl, and acetyl isoflavones and a corresponding increase in the concentrations of aglycone isoflavones, with the magnitude of these changes increasing with increasing storage temperature and moisture content. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Raster image correlation spectroscopy (RICS) for measuring fast protein dynamics and concentrations with a commercial laser scanning confocal microscope.

    Science.gov (United States)

    Brown, C M; Dalal, R B; Hebert, B; Digman, M A; Horwitz, A R; Gratton, E

    2008-01-01

    Raster image correlation spectroscopy (RICS) is a new and novel technique for measuring molecular dynamics and concentrations from fluorescence confocal images. The RICS technique extracts information about molecular dynamics and concentrations from images of living cells taken on commercial confocal systems. Here we develop guidelines for performing the RICS analysis on an analogue commercial laser scanning confocal microscope. Guidelines for typical instrument settings, image acquisition settings and analogue detector characterization are presented. Using appropriate instrument/acquisition parameters, diffusion coefficients and concentrations can be determined, even for highly dynamic dye molecules in solution. Standard curves presented herein demonstrate the ability to detect protein concentrations as low as approximately 2 nM. Additionally, cellular measurements give accurate values for the diffusion of paxillin-enhanced-green fluorescent protein (EGFP), an adhesion adaptor molecule, in the cytosol of the cell and also show slower paxillin dynamics near adhesions where paxillin interacts with immobile adhesion components. Methods are presented to account for bright immobile structures within the cell that dominate spatial correlation functions; allowing the extraction of fast protein dynamics within and near these structures. A running average algorithm is also presented to address slow cellular movement or movement of cellular features such as adhesions. Finally, methods to determine protein concentration in the presence of immobile structures within the cell are presented. A table is presented giving guidelines for instrument and imaging setting when performing RICS on the Olympus FV300 confocal and these guidelines are a starting point for performing the analysis on other commercial confocal systems.

  9. Cadmium Induces the Activation, Concentration, and Expression of MTF-1 Protein As a Biomolecular Response of Sea Urchins Deadema setosum

    Directory of Open Access Journals (Sweden)

    Dominggus Rumahlatu

    2017-12-01

    Full Text Available Heavy metal cadmium has toxic effects in the bodies of living organisms.However, the bodies of living organisms have the ability to regulate a wide range of toxic entering the body by using biomolecular response that aims at maintaining cell homeostasis. In this research an induction of cadmium in sea urchins Deadema setosum was carried out. Cadmium was induced in 6 levels of concentration in six treatment groups of D. setosum. Each group consisted of 7 individuals.The examination of MTF-1 protein activation was carried out by immunohistochemical method, the measurement of MTF-1 protein concentration was carried out by the indirect ELISA method (Enzyme Linked Imunoasssay and the examination of MTF-1 protein expression was carried out by western blotting method. The data were analyzed using descriptive analysis and One Way ANOVA followed by Duncan test 0.05 to determine the mean difference of MTF-1 protein concentration of D. setosum. The research results showed that cadmium induction activated MTF-1 characterized by the condition of liver cells of D. setosum which was brown.The higher the exposure of cadmium concentration on D. setosum, the higher the concentration of MTF-1 expressed

  10. May salivary gland secretory proteins from hematophagous leeches (Hirudo verbana reach pharmacologically relevant concentrations in the vertebrate host?

    Directory of Open Access Journals (Sweden)

    Sarah Lemke

    Full Text Available Saliva of hematophagous leeches (Hirudo sp. contains bioactive proteins which allow the leech proper feeding and storage of ingested blood, but may also exert effects in the host. Leech therapy is used to treat many different ailments in humans, although only a small fraction of salivary proteins are characterized yet. Moreover, we do not know whether complete transfer of salivary proteins stored in the unicellular salivary glands in a leech to the host during feeding may generate concentrations that are sufficiently high to affect physiological processes in the host. Our 3D reconstruction of a portion of internal leech tissue from histological sections revealed that one leech contains approx. 37,000 salivary gland cells. Using tissue slices from pig liver and mouse skeletal muscle for reference, we obtained data for protein densities in leech salivary gland cells. As individual salivary cells are voluminous (67,000 µm(3 and the stored proteins are densely packed (approx. 500 µg/mm(3, we extrapolated that a single leech may contain up to 1.2 mg of salivary proteins. Analyzing protein extracts of unfed or fed leeches by 2D electrophoresis, we calculated the relative molar amounts of individual salivary proteins in the mass range of 17-60 kDa which may be released from a single leech during feeding. Distribution of these salivary proteins in the host (assumed plasma volume of 5 l may result in concentrations of individual compounds between 3 and 236 pmol/l. Such concentrations seem sufficiently high to exert biochemical interactions with target molecules in the host.

  11. 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....

  12. Effects ofEIiergy Intake and, Dietary Protein Concentration on ~r ...

    African Journals Online (AJOL)

    ature of rats giYen 'a low protein dieL However, low MEm vaiue 'tha(coinpute'd, by "mUltiple re- given that animals fed on .high protein diet depos- gression equatiOl)S' for anirhals fed on high pro- ited protein at a relatively higher rate and protein tein diet in the present study, , . . energy accounted :ror 0.71 of the total energy ...

  13. 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...

  14. 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.......7%, respectively (considered moderately accurate). Conclusions and Clinical Relevance-Although serum CRP concentration was associated with outcome in puppies with canine parvovirus enteritis, it did not prove to be a good predictor of outcome when used alone....

  15. 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.

  16. Heat Shock Protein member A2 forms a stable complex with angiotensin converting enzyme and protein disulfide isomerase A6 in human spermatozoa.

    Science.gov (United States)

    Bromfield, Elizabeth G; McLaughlin, Eileen A; Aitken, Robert John; Nixon, Brett

    2016-02-01

    Given the importance of the chaperone Heat Shock Protein A2 (HSPA2) in the regulation of male fertility, this study aimed to identify and characterize additional proteins that may rely on the activity of this chaperone in human spermatozoa. In view of the findings in this study we propose that angiotensin converting enzyme (ACE) and protein disulfide isomerase A6 (PDIA6) are novel interacting proteins of HSPA2 and that this multimeric complex may participate in key elements of the fertilization cascade. The molecular chaperone HSPA2 plays a pivotal role in the remodelling of the sperm surface during capacitation. Indeed, human spermatozoa that are deficient in HSPA2 protein expression lack the ability to recognize human oocytes, resulting in repeated IVF failure in a clinical setting. Moreover, our recent work has shown that defective HSPA2 function induced by oxidative stress leads to the aberrant surface expression of one of its interacting proteins, arylsulfatase A, and thus contributes to a loss of sperm-zona pellucida adhesion. Human spermatozoa were collected from fertile donors, capacitated and prepared for Blue Native Polyacrylamide Gel Electrophoresis (BN-PAGE) analysis. Protein complexes resolved via BN-PAGE were excised and their constituents were identified using mass spectrometry. The interactions between ACE, PDIA6 and HSPA2 were then confirmed using immunoprecipitation and proximity ligation assays and the localization of these proteins was assessed in isolated spermatozoa and commercially available human testis tissue sections. Finally, pharmacological inhibition of ACE was performed to assess the role of ACE in human sperm capacitation. Herein we have identified ACE and PDIA6 as potential HSPA2-interacting proteins and shown that this assemblage resides in membrane raft microdomains located in the peri-acrosomal region of the sperm head. Additionally, the surface expression of PDIA6, but not ACE, was shown to be dynamically regulated during sperm

  17. 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)

  18. Chemical composition, molecular weight distribution, secondary structure and effect of NaCl on functional properties of walnut (Juglans regia L) protein isolates and concentrates.

    Science.gov (United States)

    Mao, Xiao-Ying; Hua, Yu-Fei

    2014-08-01

    Chemical composition, molecular weight distribution, secondary structure and effect of sodium chloride concentration on functional properties of walnut protein isolates, concentrates and defatted walnut flour were study. Compared with walnut protein concentrates (75.6%) and defatted walnut flour (52.5%), walnut protein isolates contain a relatively high amount of protein (90.5%). The yield of walnut protein isolates and concentrates was 43.2% and 76.6%, respectively. In molecular weight distribution study, Walnut protein isolates showed one peak with molecular weight of 106.33 KDa (100%) and walnut protein concentrates showed four peaks with molecular weight of 16,725 KDa (0.8%),104.943 KDa(63.9%), 7.3 KDa (11.4%), 2.6 KDa (23.9%). The secondary structure of walnut protein isolates was similar to that of walnut protein concentrates, but was differ from that of defatted walnut flour. The addition of sodium chloride (0 ~ 1 M) could improve the functionality of walnut protein concentrates, isolates and defatted walnut flour. The maximum solubility, water absorption capacity, emulsifying properties and foaming properties of walnut protein isolates, concentrates and defatted walnut flour were at sodium chloride solutions of 1.0 M, 0.6 M, 0.4 M, 0.6 M, respectively. The solubility of walnut protein concentrates (32.5%) in distilled water with 0 M sodium chloride was lower than that of walnut protein isolates (35.2%). The maximum solubility of walnut protein isolates, concentrates and defatted walnut flour in solution were 36.8%, 33.7% and 9.6% at 1.0 M sodium chloride solutions, respectively. As compared with other vegetable proteins, walnut protein isolates and concentrates exhibited better emulsifying properties and foam stability.

  19. Protein requirements for females of Nellore, Nellore × Angus and Nellore × Simmental fed on two forage: concentrate ratios

    Directory of Open Access Journals (Sweden)

    Evaristo Jorge Oliveira de Souza

    2012-03-01

    Full Text Available This study aimed to determine the protein requirements for females of Nellore, F1 Nellore × Angus and F1 Nellore × Simmental fed on two concentrate levels (30 and 50%. Sixty heifers from three genetic groups with 18 months of age were used: 20 Nellore, 20 Nellore × Angus and 20 Nellore × Simmental. Twelve heifers of the reference group (four of each genetic group were slaughtered at the beginning of the experiment. Another 12 heifers (four of each genetic group were fed on the level of maintenance and 36 heifers (12 animals of each genetic group were kept in power system ad libitum with 30% (six of each group or 50% (six of each group dietary dry matter in concentrate. Heifers were randomly assigned to six treatments in a 3 × 2 factorial arrangement (three genetic groups and two diets with six replicates per treatment. Nine more heifers (three from each genetic group were used to estimate the apparent digestibility coefficients of food in a parallel experiment. A model was fitted according to the protein retained as function of the gain of empty body weight (EBW and retained energy (RE to calculate the protein net requirements. To estimate the metabolizable protein requirements for maintenance the consumption of metabolizable protein was contrasted with EBW. The joint use of the equation net protein gain (NPG = 197.40 × EBWg - 11.14 × RE is recommended to predict the protein net requirements for weight gain. Protein and metabolizable protein net requirements for maintenance are 1.07 and 3.88 g/EBW0.75/day, respectively. The use efficiency of metabolizable protein for gain of all genetic groups is 37.04%.

  20. Stable transfection of protein kinase C alpha cDNA in hormone-dependent breast cancer cell lines

    OpenAIRE

    Tonetti, D A; Chisamore, M J; Grdina, W; Schurz, H; Jordan, V C

    2000-01-01

    An inverse relationship between protein kinase C (PKC) activity and oestrogen receptor (ER) expression in human breast cell lines and tumours has been firmly established over the past 10 years. To determine whether specific alterations in PKC expression accompany hormone-independence, we examined the expression of PKC isozymes in the hormone-independent human breast cancer cell clones MCF-7 5C and T47D:C42 compared with their hormone-dependent counterparts, MCF-7 A4, MCF-7 WS8 and T47D:A18 re...

  1. A repeat protein links Rubisco to form the eukaryotic carbon-concentrating organelle.

    Science.gov (United States)

    Mackinder, Luke C M; Meyer, Moritz T; Mettler-Altmann, Tabea; Chen, Vivian K; Mitchell, Madeline C; Caspari, Oliver; Freeman Rosenzweig, Elizabeth S; Pallesen, Leif; Reeves, Gregory; Itakura, Alan; Roth, Robyn; Sommer, Frederik; Geimer, Stefan; Mühlhaus, Timo; Schroda, Michael; Goodenough, Ursula; Stitt, Mark; Griffiths, Howard; Jonikas, Martin C

    2016-05-24

    Biological carbon fixation is a key step in the global carbon cycle that regulates the atmosphere's composition while producing the food we eat and the fuels we burn. Approximately one-third of global carbon fixation occurs in an overlooked algal organelle called the pyrenoid. The pyrenoid contains the CO2-fixing enzyme Rubisco and enhances carbon fixation by supplying Rubisco with a high concentration of CO2 Since the discovery of the pyrenoid more that 130 y ago, the molecular structure and biogenesis of this ecologically fundamental organelle have remained enigmatic. Here we use the model green alga Chlamydomonas reinhardtii to discover that a low-complexity repeat protein, Essential Pyrenoid Component 1 (EPYC1), links Rubisco to form the pyrenoid. We find that EPYC1 is of comparable abundance to Rubisco and colocalizes with Rubisco throughout the pyrenoid. We show that EPYC1 is essential for normal pyrenoid size, number, morphology, Rubisco content, and efficient carbon fixation at low CO2 We explain the central role of EPYC1 in pyrenoid biogenesis by the finding that EPYC1 binds Rubisco to form the pyrenoid matrix. We propose two models in which EPYC1's four repeats could produce the observed lattice arrangement of Rubisco in the Chlamydomonas pyrenoid. Our results suggest a surprisingly simple molecular mechanism for how Rubisco can be packaged to form the pyrenoid matrix, potentially explaining how Rubisco packaging into a pyrenoid could have evolved across a broad range of photosynthetic eukaryotes through convergent evolution. In addition, our findings represent a key step toward engineering a pyrenoid into crops to enhance their carbon fixation efficiency.

  2. 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.

  3. Consequences of protein supplementation for anorexia, expression of immunity and plasma leptin concentrations in parasitized ewes of two breeds.

    Science.gov (United States)

    Zaralis, Konstantinos; Tolkamp, Bert J; Houdijk, Jos G M; Wylie, Alastair R G; Kyriazakis, Ilias

    2009-02-01

    The periparturient relaxation of immunity (PPRI) against parasites in ewes has a nutritional basis. We investigated whether ewes experience a reduction in food intake (anorexia) during PPRI and if the magnitude of anorexia is affected by host production potential and dietary protein supplementation. We also investigated whether nematode infection is linked to plasma leptin concentrations in periparturient ewes. The experiment was a 2 x 2 x 2 factorial design. Two breeds of twin-bearing/lactating ewes (Greyface cross, G (n 32) and Scottish Blackface, B (n 32)) were used. Half of the ewes were trickle infected with 30,000 larvae of the abomasal parasite Teladorsagia circumcincta per week and the other half were not. During the experiment, all ewes had ad libitum access to a low-protein diet that provided less protein than the recommended allowance. In addition, half of the ewes received a protein supplement that resulted in protein intakes that exceeded recommendations. Nematode infection resulted in a breakdown of immunity to parasites and a reduction in food intake in both breeds. The breeds differed in the extent of PPRI (G ewes having higher faecal egg counts than B ewes), but not in the magnitude of anorexia. Protein supplementation resulted in a reduction in faecal egg counts, but had no effect on the magnitude of anorexia. Plasma leptin concentrations changed significantly over time, but were not affected by protein supplementation or infection. It is concluded that infection with T. circumcincta in periparturient ewes results in anorexia that is not alleviated by protein supplementation and seems unrelated to plasma leptin concentrations.

  4. Cell age dependent concentration of Escherichia coli divisome proteins analyzed with ImageJ and ObjectJ

    Directory of Open Access Journals (Sweden)

    Norbert O.E. Vischer

    2015-06-01

    Full Text Available The rod-shaped Gram-negative bacterium Escherichia coli multiplies by elongation followed by binary fission. Longitudinal growth of the cell envelope and synthesis of the new poles are organized by two protein complexes called elongasome and divisome, respectively. We have analyzed the spatio-temporal localization patterns of many of these morphogenetic proteins by immunolabeling the wild type strain MC4100 grown to steady state in minimal glucose medium at 28°C. This allowed the direct comparison of morphogenetic protein localization patterns as a function of cell age as imaged by phase contrast and fluorescence wide field microscopy. Under steady state conditions the age distribution of the cells is constant and is directly correlated to cell length. To quantify cell size and protein localization parameters in thousands of labeled cells, we developed 'Coli-Inspector', which is a project running under ImageJ with the plugin 'ObjectJ'. ObjectJ organizes image-analysis tasks using an integrated approach with the flexibility to produce different output formats from existing markers such as intensity data and geometrical parameters. ObjectJ supports the combination of automatic and interactive methods giving the user complete control over the method of image analysis and data collection, with visual inspection tools for quick elimination of artifacts. Coli-inspector was used to sort the cells according to division cycle cell age and to analyze the spatio-temporal localization pattern of each protein. A unique dataset has been created on the concentration and position of the proteins during the cell cycle. We show for the first time that a subset of morphogenetic proteins have a constant cellular concentration during the cell division cycle whereas another set exhibits a cell division cycle dependent concentration variation. Using the number of proteins present at midcell, the stoichiometry of the divisome is discussed.

  5. The Concentration and Yield of Hordein and some Lysine-Rich Proteins as Influenced by the lys gene of Hiproly Barley

    DEFF Research Database (Denmark)

    Balasaraswathi, R.; Køie, B; Doll, Hans

    1984-01-01

    the corresponding normal gene. The lys lines had 16% lower grain yield and 13% lower single seed weight than the corresponding normal lines. The concentration of hordein in the protein and the yield of this storage protein were strongly reduced in the lys lines. On the contrary, the concentration of protein Z, β...

  6. Variation of protein MWD parameters and their associations with free asparagine concentration and quality characteristics in hard red spring wheat

    Science.gov (United States)

    This research was performed to determine variation of protein molecular weight distribution (MWD) parameters using size exclusion HPLC and their associations with quality characteristics and free asparagine concentration (FAC) using eleven hard red spring (HRS) wheat genotypes grown at three locatio...

  7. Atomic detail brownian dynamics simulations of concentrated protein solutions with a mean field treatment of hydrodynamic interactions.

    Energy Technology Data Exchange (ETDEWEB)

    Mereghetti, Paolo; Wade, Rebecca C.

    2012-07-26

    High macromolecular concentrations are a distinguishing feature of living organisms. Understanding how the high concentration of solutes affects the dynamic properties of biological macromolecules is fundamental for the comprehension of biological processes in living systems. In this paper, we describe the implementation of mean field models of translational and rotational hydrodynamic interactions into an atomically detailed many-protein brownian dynamics simulation method. Concentrated solutions (30-40% volume fraction) of myoglobin, hemoglobin A, and sickle cell hemoglobin S were simulated, and static structure factors, oligomer formation, and translational and rotational self-diffusion coefficients were computed. Good agreement of computed properties with available experimental data was obtained. The results show the importance of both solvent mediated interactions and weak protein-protein interactions for accurately describing the dynamics and the association properties of concentrated protein solutions. Specifically, they show a qualitative difference in the translational and rotational dynamics of the systems studied. Although the translational diffusion coefficient is controlled by macromolecular shape and hydrodynamic interactions, the rotational diffusion coefficient is affected by macromolecular shape, direct intermolecular interactions, and both translational and rotational hydrodynamic interactions.

  8. Effects of tylosin administration on C-reactive protein concentration and carriage of Salmonella enterica in pigs.

    Science.gov (United States)

    Kim, Hyeun Bum; Singer, Randall S; Borewicz, Klaudyna; White, Bryan A; Sreevatsan, Srinand; Johnson, Timothy J; Espejo, Luis A; Isaacson, Richard E

    2014-05-01

    To evaluate the effects of tylosin on C-reactive protein concentration, carriage of Salmonella enterica, and antimicrobial resistance genes in commercial pigs. 120 pigs on 2 commercial farms. A cohort of sixty 10-week-old pigs in 4 pens/farm (15 pigs/pen) was randomly selected. Equal numbers of pigs were given feed containing tylosin (40 μg/g of feed) for 0, 6, or 12 weeks. C-reactive protein concentrations were measured, microbial culture for S enterica in feces was performed, and antimicrobial resistance genes in feces were quantified. No significant associations were detected between C-reactive protein concentration or S enterica status and tylosin treatment. During the 12 weeks of tylosin administration, increased levels of 6 antimicrobial resistance genes did not occur. Treatment of pigs with tylosin did not affect C-reactive protein concentration or reduce carriage or load of S enterica. There was no evidence that pigs receiving tylosin had increased carriage of the 6 antimicrobial resistance genes measured. S enterica is a public health concern. Use of the antimicrobial growth promoter tylosin did not pose a public health risk by means of increased carriage of S enterica.

  9. 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...

  10. 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

  11. Effect of neutrase, alcalase, and papain hydrolysis of whey protein concentrates on iron uptake by Caco-2 cells

    NARCIS (Netherlands)

    Ou, K.Q.; Liu, Y.Z.; Zhang, L.B.; Yang, X.G.; Huang, Z.W.; Nout, M.J.R.; Liang, J.

    2010-01-01

    Effects of enzymatic hydrolysates of whey protein concentrates (WPC) on iron absorption were studied using in vitro digestion combined with Caco-2 cell models for improved iron absorption. Neutrase- and papain-treated WPC could improve iron absorption; especially hydrolysates by Neutrase could

  12. 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

  13. 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

  14. MaXIC-Q Web: a fully automated web service using statistical and computational methods for protein quantitation based on stable isotope labeling and LC-MS.

    Science.gov (United States)

    Tsou, Chih-Chiang; Tsui, Yin-Hao; Yian, Yi-Hwa; Chen, Yi-Ju; Yang, Han-Yin; Yu, Chuan-Yih; Lynn, Ke-Shiuan; Chen, Yu-Ju; Sung, Ting-Yi; Hsu, Wen-Lian

    2009-07-01

    Isotope labeling combined with liquid chromatography-mass spectrometry (LC-MS) provides a robust platform for analyzing differential protein expression in proteomics research. We present a web service, called MaXIC-Q Web (http://ms.iis.sinica.edu.tw/MaXIC-Q_Web/), for quantitation analysis of large-scale datasets generated from proteomics experiments using various stable isotope-labeling techniques, e.g. SILAC, ICAT and user-developed labeling methods. It accepts spectral files in the standard mzXML format and search results from SEQUEST, Mascot and ProteinProphet as input. Furthermore, MaXIC-Q Web uses statistical and computational methods to construct two kinds of elution profiles for each ion, namely, PIMS (projected ion mass spectrum) and XIC (extracted ion chromatogram) from MS data. Toward accurate quantitation, a stringent validation procedure is performed on PIMSs to filter out peptide ions interfered with co-eluting peptides or noise. The areas of XICs determine ion abundances, which are used to calculate peptide and protein ratios. Since MaXIC-Q Web adopts stringent validation on spectral data, it achieves high accuracy so that manual validation effort can be substantially reduced. Furthermore, it provides various visualization diagrams and comprehensive quantitation reports so that users can conveniently inspect quantitation results. In summary, MaXIC-Q Web is a user-friendly, interactive, robust, generic web service for quantitation based on ICAT and SILAC labeling techniques.

  15. 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

  16. Characterization of the nutritional quality of amaranth leaf protein concentrates and suitability of fish meal replacement in Nile tilapia feeds

    Directory of Open Access Journals (Sweden)

    Charles C. Ngugi

    2017-02-01

    Full Text Available A number of leafy vegetables, their protein concentrates and hydrolasates are under evaluation as alternative protein ingredients to fish meal (FM in aquafeeds. This study evaluated the nutritional characteristics and suitability of replacing FM with the amaranth (Amaranthus hybridus leaf protein concentrates (ALPC as a protein ingredient in the diet of Nile tilapia (Oreochromis niloticus. Experimental diets were formulated, where 100%, 75%, 50%, 40%, 20% and 0% FM protein was substituted by protein from ALPC. The six dietary treatments were tested in triplicate in static flow-through tanks. The substitution effects were compared in terms of fish growth performance, nutrient utilization, whole body composition and apparent nutrient digestibility. After 160 days of feeding, the growth, nutrient utilization and Feed Conversion Ratio (FCR in fish fed diets containing 100%, 75%, 50%, 40% and 20% FM were better (P < 0.05 than those fed diet with 0% FM. The Apparent nutrient digestibility was high for protein, lipid and energy and differed significantly among the dietary treatments (P < 0.05. Protein digestibility in fish was highest in feed formulated with 100%, 75%, 50% and 40% FM, which were significantly (P < 0.05 higher than at 25% and 0% FM. Lipid digestibility was comparable for all the diets except fish fed 0% FM. Digestible carbohydrates and dry matter were similar for all dietary treatments (P < 0.05. We demonstrate that it is possible to replace up to 80% of fish meal with ALPC without compromising the performance O. niloticus. These results demonstrate that although it is possible to replace large part of fish meal with ALPC, it is not possible to eliminate it in Nile tilapia diet as alternative protein ingredient.

  17. Short communication: Annatto in Cheddar cheese-derived whey protein concentrate is primarily associated with milk fat globule membrane.

    Science.gov (United States)

    Zhu, D; Damodaran, S

    2012-02-01

    The yellow color of Cheddar cheese whey arises from a residual amount of annatto that partitions into the whey during Cheddar cheese manufacture. Bleaching of the color using hydrogen peroxide or benzoyl peroxide is often a prerequisite to produce an acceptable neutral-colored whey protein concentrate and isolate. However, the use of these strong oxidizing agents often generates off-flavors as a result of lipid oxidation and results in loss of nutritive value due to protein oxidation. The objective of this study was to determine the extent of partitioning of annatto between protein, milk fat globule membrane (MFGM), and aqueous (serum) phases of cheese whey so that a simple method can be developed to remove annatto from cheese whey. The MFGM was separated from Cheddar cheese whey using a recently developed novel method. Quantitative analysis of the distribution of annatto in the fat-free whey protein isolate (WPI), the MFGM fractions, and the serum phase revealed that annatto was not bound to the protein fraction but was mostly distributed between the serum phase and the MFGM fraction. The results showed that a colorless WPI or whey protein concentrate could be produced from Cheddar cheese whey by separation of MFGM from the whey, followed by diafiltration. This approach will negate the need for using bleaching agents. Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  18. Protein profile and alpha-lactalbumin concentration in the milk of standard and transgenic goats expressing recombinant human butyrylcholinesterase.

    Science.gov (United States)

    Baldassarre, H; Schirm, M; Deslauriers, J; Turcotte, C; Bordignon, V

    2009-08-01

    The expression of recombinant proteins of pharmaceutical interest in the milk of transgenic farm animals can result in phenotypes exhibiting compromised lactation performance, as a result of the extraordinary demand placed on the mammary gland. In this study, we investigated differences in the protein composition of milk from control and transgenic goats expressing recombinant human butyrylcholinesterase. In Experiment 1, the milk was characterized by gel electrophoresis and liquid chromatography/mass spectrometry in order to identify protein bands that were uniquely visible in the transgenic milk and/or at differing band densities compared with controls. Differences in protein content were additionally evaluated by computer assisted band densitometry. Proteins identified in the transgenic milk only included serum proteins (i.e. complement component 3b, ceruloplasmin), a cytoskeleton protein (i.e. actin) and a stress-induced protein (94 kDA glucose-regulated protein). Proteins exhibiting evident differences in band density between the transgenic and control groups included immunoglobulins, serum albumin, beta-lactoglobulin and alpha-lactalbumin. These results were found to be indicative of compromised epithelial tight junctions, premature mammary cell death, and protein synthesis stress resulting from transgene expression. In Experiment 2, the concentration of alpha-lactalbumin was determined using the IDRing assay and was found to be significantly reduced on day 1 of lactation in transgenic goats (4.33 +/- 0.97 vs. 2.24 +/- 0.25 mg/ml, P 0.05). We concluded that a decreased/delayed expression of the alpha-lactalbumin gene may be the cause for the delayed start of milk production observed in this herd of transgenic goats.

  19. Electrophysiological Features of Single Store-Operated Calcium Channels in HEK S4 Cell Line with Stable STIM1 Protein Knockdown.

    Science.gov (United States)

    Shalygin, A V; Vigont, V A; Glushankova, L N; Zimina, O A; Kolesnikov, D O; Skopin, A Yu; Kaznacheeva, E V

    2017-07-01

    An important role in intracellular calcium signaling is played by store-operated channels activated by STIM proteins, calcium sensors of the endoplasmic reticulum. In stable STIM1 knockdown HEK S4 cells, single channels activated by depletion of intracellular calcium stores were detected by cell-attached patch-clamp technique and their electrophysiological parameters were described. Comparison of the properties of single channels in HEK293 and HEK S4 cells revealed no significant differences in their current-voltage curves, while regulation of store-operated calcium channels in these cell lines depended on the level of STIM1 expression. We can conclude that electrophysiological peculiarities of store-regulated calcium entry observed in different cells can be explained by differences in STIM1 expression.

  20. Analysis of the thermal energy requirements for the extraction of leaf protein concentrate from some green plants

    Energy Technology Data Exchange (ETDEWEB)

    Tangka, Julius K. [Dschang Univ., Dept. of Agricultural Engineering, Dschang (Cameroon)

    2003-12-01

    Extraction of protein from the leaves of green plants is very important because of the high cost of conventional forms of protein such as meat, milk and fish. In order to design machinery for this extraction, and also to embark on leaf protein concentrate extraction, it is necessary to measure and analyse the energy requirements to carry out each process involved in the extraction, using different plant species. Experiments were carried out to determine the amount of crude protein, and the thermal energy required to extract leaf protein concentrate, from juices obtained from the leaves of some plant species. Leaves from the following plants were selected: cassava (Manihot esculanta), Siam weed (Chromolaena odorata), bitter leaf (Vernonia amygdalina), gliricidia (Gliricidia maculata) and thorny tree (Hura crepetans). The leaves from the plant species were macerated in a laboratory pulper. Juice was obtained from the samples using perforated cylinders and a hydraulic press. The specific heat capacity of the juices was determined using the cooling curve method. The values of the heat capacities were used to calculate the amount of thermal energy required to raise the temperature of each juice from its normal temperature of about 25 deg C to a total protein coagulation temperature of about 80 deg C. The crude protein content of the extract was determined using the Kjeldal method. Results indicate that the green coagulum extracted from all the juices all have a protein content of at least 37%. The thermal energy required to coagulate protein from the juices ranges from 1.59 kJ kg{sup -1} for Hura crepetans to 2.7 kJ kg{sup -1} for Vernonia amygdalina. The energy requirement to obtain crude protein (CP) ranges from 8 kJ kg{sup -1} [CP] with Bura crepetans to 182 kJ kg{sup -1} [CP] with Vernonia amygdalina. Both results are statistically significant at the 0.01 confidence interval. It is concluded that the choice of plant species can significantly lower the thermal energy

  1. 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.

  2. 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-...

  3. 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.

  4. 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

  5. Can serum concentration of C-reactive protein, albumin and body ...

    African Journals Online (AJOL)

    The C-reactive protein was estimated using semi-quantitative method, albumin level was estimated using bromocresol green method, and biuret method was used for total protein. The CD4+T-cell count of the subjects was determined using CyFlow Analyzer while their weights were measured using high precision weighing ...

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

    African Journals Online (AJOL)

    However, the linear and quadratic curves and coefficients revealed substantial increase in total protein content in all the brain regions from day-old to between 12 and 20 months, thereafter declining rapidly and, in certain regions, gradually to much lower levels. In the pons, a rise in total protein of 1,091% was observed from ...

  7. 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

  8. Ileal digestibility of amino acids of unheated and autoclaved pea protein concentrate in broilers.

    Science.gov (United States)

    Frikha, M; Valencia, D G; de Coca-Sinova, A; Lázaro, R; Mateos, G G

    2013-07-01

    The effects of autoclaving 2 varieties of micronized (fine grinding) pea protein concentrate (PPC) on the ileal digestibility (ID) of CP and amino acids (AA) were studied in broilers. There was a control diet based on fermented soybean meal (FSBM) and 4 extra diets in which the FSBM was substituted on a CP basis by PPC from 2 different pea cultivars (PPC-1 and PPC-2), either unheated or autoclaved. Chicks were fed a common diet from 1 to 17 d of age and, then, their respective experimental diets from 18 to 21 d of age. Each treatment was replicated 6 times. Autoclaving reduced trypsin inhibitor activity (TIA) but had little effect on the saponin content of the PPC. The apparent ID (AID) of CP was similar for the FSBM and the unheated PPC and lower for both than for the autoclaved PPC. Autoclaving improved (P < 0.001) the AID of CP (87.6 vs. 82.2%) and most indispensable AA (e.g., 92.1 vs. 88.8% for Lys and 83.6 vs. 76.5% for Thr) of the PPC. The improvement in CP and AA digestibility with autoclaving varied with the PPC used and was consistent with the reduction in TIA observed (9.4 to 2.8 mg/g for PPC-1 vs. 9.1 to 5.3 mg/g for PPC-2). The standardized ID (SID) of most indispensable AA was similar for the FSBM and the PPC-2 and higher for both than for the PPC-1 (P < 0.05). For Lys, the lowest SID value was observed for the FSBM and the highest for the PPC-2 either unheated or autoclaved. It is concluded that the ID of the AA of the PPC improved with heating and was in general higher for the autoclaved PPC than for the FSBM. Consequently, heat processed PPC is a good alternative to FSBM and unheated PPC in starter diets for broilers.

  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. QTL identification of grain protein concentration and its genetic correlation with starch concentration and grain weight using two populations in maize (Zea mays L.).

    Science.gov (United States)

    Li, Yuling; Wang, Yanzhao; Wei, Mengguan; Li, Xuehui; Fu, Jiafeng

    2009-04-01

    Protein is one of the three main storage chemical components in maize grains, and is negatively correlated with starch concentration (SC). Our objective was to analyse the influence of genetic backgrounds on QTL detection for protein concentration (PC) and to reveal the molecular genetic associations between PC and both SC and grain weight (GWP). Two hundred and eighty-four (Pop1) and 265 (Pop2) F(2:3) families were developed from two crosses between one high-oil maize inbred GY220 and two normal maize inbreds 8984 and 8622 respectively, and were genotyped with 185 and 173 pairs of SSR markers. PC, SC and GWP were evaluated under two environments. Composite interval mapping (CIM) and multiple interval mapping (MIM) methods were used to detect single-trait QTL for PC, and multiple-trait QTL for PC with both SC and GWP. No common QTL were shared between the two populations for their four and one PC QTL. Common QTL with opposite signs of effects for PC and SCGWP were detected on three marker intervals at bins 6.07-6.08, 8.03 and 8.03-8.04. Multiple-traits QTL mapping showed that tightly-linked QTL, pleiotropic QTL and QTL having effects with opposite directions for PC and SCGWP were all observed in Pop1, while all QTL reflected opposite effects in Pop2.

  12. 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

  13. 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.

  14. Maternal Low Quality Protein Diet Alters Plasma Amino Acid Concentrations of Weaning Rats

    Directory of Open Access Journals (Sweden)

    Arzu Kabasakal Cetin

    2015-12-01

    Full Text Available Several studies have indicated the influence of a maternal low protein diet on the fetus. However, the effect of a maternal low quality protein diet on fetal growth and development is largely unknown. Wistar rats (11 weeks old were mated and maintained on either a chow diet with 20% casein (n = 6 as the control group (C, or a low quality protein diet with 20% wheat gluten (n = 7 as the experimental group (WG through gestation and lactation. Maternal body weights were similar in both groups throughout the study. Birth weights were not influenced by maternal diet and offspring body weights during lactation were similar between the groups. Offspring’s plasma amino acid profiles showed that plasma methionine, glutamine and lysine were significantly lower and aspartic acid, ornithine and glycine-proline were significantly higher in the WG. Plant based protein comprises an important part of protein intake in developing countries. It is well-known that these diets can be inadequate in terms of essential amino acids. The current study shows differential effects of a maternal low quality protein diet on the offspring’s plasma amino acids. Future studies will examine further aspects of the influence of maternal low quality protein diets on fetal growth and development.

  15. 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.

  16. 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).

  17. 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.

  18. 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

  19. 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.

  20. Testicular superoxide dismutase activity, heat shock protein 70 concentration and blood plasma inhibin-alpha concentration of dogs with a Sertoli cell tumor in a unilateral cryptorchid testis.

    Science.gov (United States)

    Kawakami, Eiichi; Hirano, Taichi; Hori, Tatsuya; Tsutsui, Toshihiko

    2007-12-01

    The proportions of Sertoli cell tumor (SCT), seminoma and Leydig cell tumor in 50 dogs with unilateral testicular tumors were 52%, 36% and 12%, respectively. The rate of occurrence of SCT in the cryptorchid testis was very high (71%). The testicular superoxide dismutase (SOD) activity, testicular heat shock protein (HSP) 70 concentration and peripheral blood plasma inhibin (INH)-alpha concentration of 10 dogs with a unilateral cryptorchid testis and no testicular tumors, 10 dogs with SCT in a unilateral cryptorchid testis and 10 normal dogs, all aged 5-15 years, were measured in order to identify high risk factors for the occurrence of SCT in the canine cryptorchid testis. The mean SOD activity in cryptorchid testes and SCTs was significantly lower and higher, respectively, than in normal testes (both Pdogs was significantly lower and higher, respectively, than in normal dogs (Pdogs and high HSP 70 concentration in the SCTs may be related to the occurrence of SCT and tumor cell proliferation in canine cryptorchid testes.

  1. 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...... in PAPP-A concentration to repeated injections of unfractionated heparin. DESIGN AND METHODS: Twenty-five healthy, non-pregnant volunteers divided into five groups (determined by gender and age) received 5000 IU unfractionated heparin intravenously. Five young men received an additional 5000 IU after 90...... and 180 min. Blood samples to determine PAPP-A concentration and APTT were drawn at different time points. RESULTS: Injection of heparin elicited increase in and rapid normalization of PAPP-A concentrations in all subjects. The group of 20-30-year-old never-pregnant women had lower responses than...

  2. 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...

  3. 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

  4. 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

  5. Orthokinetic flocculation of caseinate-stabilized emulsions : influence of calcium concentration, shear rate and protein content

    NARCIS (Netherlands)

    Schokker, E.P.; Dalgleish, D.G.

    2000-01-01

    Calcium-induced flocculation of caseinate-stabilized soybean oil-in- water emulsions in conditions of Couette flow was studied. A concentrated emulsion (20% oil, 0.5-2.0% sodium caseinate in 20 mM imidazole, pH 7) was diluted 20 times in buffer containing concentrations of CaCl2 between 9 and 17 mM

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

    DEFF Research Database (Denmark)

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

    2017-01-01

    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...... 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....

  7. The use of stable and unstable green fluorescent proteins for studies in two bacterial models: Agrobacterium tumefaciens and Xanthomonas campestris pv. campestris.

    Science.gov (United States)

    Sabuquillo, Pilar; Gea, Adela; Matas, Isabel M; Ramos, Cayo; Cubero, Jaime

    2017-05-01

    Fluorescent proteins have been used to track plant pathogens to understand their host interactions. To be useful, the transgenic pathogens must present similar behaviour than the wild-type isolates. Herein, a GFP marker was used to transform two plant pathogenic bacteria, Agrobacterium and Xanthomonas, to localize and track the bacteria during infection. The transgenic bacteria were evaluated to determine whether they showed the same fitness than the wild-type strains or whether the expression of the GFP protein interfered in the bacterial activity. In Agrobacterium, the plasmid used for transformation was stable in the bacteria and the strain kept the virulence, while Xanthomonas was not able to conserve the plasmid and transformed strains showed virulence variations compared to wild-type strains. Although marking bacteria with GFP to track infection in plants is a common issue, works to validate the transgenic strains and corroborate their fitness are not usual. Results, presented here, confirm the importance of proper fitness tests on the marked strains before performing localization assays, to avoid underestimation of the microbe population or possible artificial effects in its interaction with the plant.

  8. The effect of spray-drying parameters on the flavor of nonfat dry milk and milk protein concentrate 70.

    Science.gov (United States)

    Park, Curtis W; Stout, Mark A; Drake, MaryAnne

    2016-12-01

    Unit operations during production influence the sensory properties of nonfat dry milk (NFDM) and milk protein concentrate (MPC). Off-flavors in dried dairy ingredients decrease consumer acceptance of ingredient applications. Previous work has shown that spray-drying parameters affect physical and sensory properties of whole milk powder and whey protein concentrate. The objective of this study was to determine the effect of inlet temperature and feed solids concentration on the flavor of NFDM and MPC 70% (MPC70). Condensed skim milk (50% solids) and condensed liquid MPC70 (32% solids) were produced using pilot-scale dairy processing equipment. The condensed products were then spray dried at either 160, 210, or 260°C inlet temperature and 30, 40, or 50% total solids for NFDM and 12, 22, or 32% for MPC70 in a randomized order. The entire experiment was replicated 3 times. Flavor of the NFDM and MPC70 was evaluated by sensory and instrumental volatile compound analyses. Surface free fat, particle size, and furosine were also analyzed. Both main effects (30, 40, and 50% solids and 160, 210, and 260°C inlet temperature) and interactions between solids concentration and inlet temperature were investigated. Interactions were not significant. In general, results were consistent for NFDM and MPC70. Increasing inlet temperature and feed solids concentration increased sweet aromatic flavor and decreased cardboard flavor and associated lipid oxidation products. Increases in furosine with increased inlet temperature and solids concentration indicated increased Maillard reactions during drying. Particle size increased and surface free fat decreased with increasing inlet temperature and solids concentration. These results demonstrate that increasing inlet temperatures and solids concentration during spray drying decrease off-flavor intensities in NFDM and MPC70 even though the heat treatment is greater compared with low temperature and low solids. Copyright © 2016 American

  9. 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

  10. 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.

  11. 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.

  12. 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 daily gain (ADG). Concentrate level did not affect (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  0.05) the carcass characteristics and carcass gain composition. Heifers fed 80% concentrate diets had larger (P gain of bones (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.

  13. 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

    -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......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...... 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

  14. 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®

  15. Effects of dietary protein concentration and specific amino acids on body weight, body composition and feather growth in young turkeys.

    Science.gov (United States)

    Wylie, L M; Robertson, G W; Hocking, P M

    2003-03-01

    1. Two randomised block factorial experiments were conducted to investigate the relationships between the effects of dietary crude protein and specific amino acid concentrations on the relative growth of the body and feathers of young turkeys. 2. Decreasing dietary crude protein concentration from 300 to 180 g/kg in experiment 1 reduced the body and breast muscle weights of a large male line of turkeys proportionally by 0.44 and 0.52 compared with 0.19 and 0.24 in a small traditional line. 3. Decreasing dietary crude protein concentration was associated with a maximum reduction in feather weight of 0.18 and 0.24 respectively in male line and traditional turkeys. The length of the feathers in the cranial region of the breast decreased from 26 to 19mm in the traditional line compared with an increase from 14 to 25 mm in male line turkeys. 4. Decreasing dietary crude protein concentration was associated with an increase in the fat content of the feather-free carcase. Male line turkeys had a higher carcase fat and lower feather dry matter content than the traditional turkeys. 5. It was concluded that dietary crude protein was preferentially partitioned to feather rather than muscle growth in the male line in contrast to a traditional line of turkeys in which the growth of feathers and muscle were affected equally. 6. In experiment 2, the amino acids arginine, valine, methionine and tyrosine were added separately to a common basal ration (180g CP/kg) to raise their concentration to that of the control ration (260 g CP/kg). Each ration was fed ad libitum to male line turkeys from 2 to 6 weeks of age. 7. Amino acid supplementation increased body and breast muscle weights. 8. Compared with the basal ration, tyrosine was associated with a reduction in feather weight whereas valine had no effect. Supplementation with arginine and methionine resulted in increased feather weights that were similar to that of the controls. 9. It was concluded that arginine and methionine were

  16. 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.

  17. Relationship of serum bilirubin concentration to kidney function and 24-hour urine protein in Korean adults

    OpenAIRE

    Shin, Ho Sik; Jung, Yeon Soon; Rim, Hark

    2011-01-01

    Abstract Background The relationships among serum bilirubin concentration, kidney function and proteinuria have yet to be fully elucidated, nor have these relationships been investigated in Korean adults. Method We retrospectively reviewed the medical records of Korean adults who were evaluated at Kosin University Gospel Hospital (Busan, Republic of Korea) during a five-year period from January 2005 to December 2009. We evaluated the relationships among serum bilirubin concentration, estimate...

  18. A specific protein-enriched enteral formula decreases cortisolemia and improves plasma albumin and amino acid concentrations in elderly patients

    Directory of Open Access Journals (Sweden)

    Pérez de la Cruz Antonio

    2010-07-01

    Full Text Available Abstract Background Old age is associated with an involuntary and progressive but physiological loss of muscle mass. The aim of this study was to evaluate the effects of exclusive consumption for 6 months of a protein-enriched enteral diet with a relatively high content of branched-chain amino acids on albuminemia, cortisolemia, plasma amino acids, insulin resistance, and inflammation biomarkers in elderly patients. Methods Thirty-two patients from the Clinical Nutrition Outpatient Unit at our hospital exclusively consumed a protein-enriched enteral diet for 6 months. Data were collected at baseline and at 3 and 6 months on anthropometric and biochemical parameters and on plasma concentrations of amino acids, cortisol, adrenocorticotropic hormone, urea, creatinine, insulin resistance, and inflammation biomarkers. Results The percentage of patients with albumin concentration below normal cut-off values decreased from 18% to 0% by the end of the study. At 6 months, concentrations of total plasma (p = 0.008 and essential amino acids (p = 0.011, especially branched-chain amino acids (p = 0.031, were higher versus baseline values, whereas 3-methylhistidine (p = 0.001, cortisol (p = 0.001 and adrenocorticotropic hormone (p = 0.004 levels were lower. Conclusions Regular intake of specific protein-enriched enteral formula increases plasma essential amino acids, especially branched-chain amino acids, and decreases cortisol and 3-methylhistidine, while plasma urea and creatinine remain unchanged.

  19. Comparison of two automated quantitative immunoassays for the determination of C reactive protein concentrations.

    Science.gov (United States)

    Barclay, E A; Coia, J E; Kale, P C; Masterton, R G

    1994-01-01

    Two quantitative, automated methods for the determination of C reactive protein (CRP) were compared: turbidimetry (Cobas Fara II, Roche, Welwyn Garden City, UK) and fluorescence polarisation TDx, Abbott, Wokingham, UK). One hundred and twenty routine serum samples submitted for measurement of CRP were tested using both procedures. The results were compared using regression line analysis and showed a high degree of correlation (r2 = 0.99, X coefficient = 1.01, constant = 0.11). C reactive protein can be accurately measured using the automated turbidimetric method which can be recommended as an alternative to fluorescence polarisation. Images PMID:7876389

  20. A Common Polymorphism in the SFTPD Gene Influences Assembly, Function, and Concentration of Surfactant Protein D

    DEFF Research Database (Denmark)

    Leth-Larsen, Rikke; Garred, Peter; Jensenius, Henriette

    2005-01-01

    Surfactant protein D (SP-D) plays important roles in the host defense against infectious microorganisms and in regulating the innate immune response to a variety of pathogen-associated molecular pattern. SP-D is mainly expressed by type II cells of the lung, but SP-D is generally found on epithel......Surfactant protein D (SP-D) plays important roles in the host defense against infectious microorganisms and in regulating the innate immune response to a variety of pathogen-associated molecular pattern. SP-D is mainly expressed by type II cells of the lung, but SP-D is generally found...