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Sample records for acidic antigenic fractions

  1. ANTIGENICITY OF POLYPEPTIDES (POLY ALPHA AMINO ACIDS)

    PubMed Central

    Maurer, Paul H.; Gerulat, Bernard F.; Pinchuck, Paul

    1964-01-01

    A new group of synthetic random polymers of α-L-amino acids has been studied for immunogenicity. With the glutamic acid and alanine copolymers, those consisting of almost equimolar amounts of the two (G60A40 and G40A60) were effective antigens in rabbits whereas those with higher glutamic acid contents (G75A25, G90A10) were poor antigens. The substitution of alanine by valine or leucine (G75V25 and G80Leu20) produced copolymers which were poor antigens in rabbits but effective in guinea pigs. L70A30, although capable of "non-specifically" precipitating serum proteins, was shown not to be antigenic in either rabbits or guinea pigs. The introduction of alanine into glutamic acid and lysine polymers (GLA series) enhanced the immunogenicity of the terpolymers, i.e., GLA30 > GLA20 > GLA10 > GL. The mechanism by which this may be accomplished is discussed as possibly being related to the reduction of the interactions between glutamyl and lysyl residues which allows the carboxyl groups to act as strong immunogenic determinants. PMID:14176288

  2. Rapid detection of Yersinia pestis recombinant fraction 1 capsular antigen.

    PubMed

    Tsui, Pei-Yi; Tsai, Hui-Ping; Chiao, Der-Jiang; Liu, Cheng-Che; Shyu, Rong-Hwa

    2015-09-01

    Yersinia pestis, an infectious bacterium that is a causative agent of plague, a disease which has been shown to be one of the most feared in history and which has caused millions of deaths. The capsule-like fraction 1 (F1) antigen expressed by Y. pestis is a known specific marker for the identification of the bacteria; therefore, the detection of F1 is important for Y. pestis recognition. In this study, a rapid, sensitive, and specific technique, the lateral flow assay (LFA), was successfully developed to detect Y. pestis by the recombinant F1 antigen. The assay that utilized an anti-F1 polyclonal antibody (Pab) to identify the bacteria was based on a double-antibody sandwich format on a nitrocellulose membrane. With the LFA method, 50 ng/ml of recombinant F1 protein and 10(5) CFU/mL of Y. pestis could be detected in less than 10 min. This assay also showed no cross-reaction with other Yersinia spp. or with some selected capsule-producing Enterobacteriaceae strains. Furthermore, detection of Y. pestis in simulated samples has been evaluated. The detection sensitivity of Y. pestis in various matrices was 10(5) CFU/mL, which was identical to that in PBS buffer. The results obtained suggest that LFA is an excellent tool for detection of Y. pestis contamination in an environment and hence can be used to monitor plague diseases when they emerge.

  3. Vaccination of mice against Taenia taeniaeformis using antigen fractions partitioned with Triton X-114.

    PubMed

    Schnieder, T; Bøgh, H O; Lightowlers, M W; Rickard, M D

    1996-01-01

    Taenia taeniaeformis oncosphere and metacestode antigens were fractioned using Triton X-114 into insoluble, aqueous and detergent rich fractions. These fractions were analysed in SDS-PAGE and immunoblots and used in vaccination trials against infection with T. taeniaeformis in mice. Qualitative differences were apparent in the spectrum of antigens partitioning into the different detergent phases but host-prospective antigens were present in all three fractions. The presence of individual antigenic components in the phases did not correlate with the degree of protection afforded by these fractions in the vaccination trials. Host protective immunogenicity of T. taeniaeformis oncosphere and metacestode extracts may be due to multiple protective antigens which partition into the different Triton X-114 fractions.

  4. Oxidative bioactivation of abacavir in subcellular fractions of human antigen presenting cells.

    PubMed

    Bell, Catherine C; Santoyo Castelazo, Anahi; Yang, Emma L; Maggs, James L; Jenkins, Rosalind E; Tugwood, Jonathan; O'Neill, Paul M; Naisbitt, Dean J; Park, B Kevin

    2013-07-15

    Human exposure to abacavir, a primary alcohol antiretroviral, is associated with the development of immunological drug reactions in individuals carrying the HLA risk allele B*57:01. Interaction of abacavir with antigen presenting cells results in cell activation through an Hsp70-mediated Toll-like receptor pathway and the provision of T-cell antigenic determinants. Abacavir's electrophilic aldehyde metabolites are potential precursors of neoantigens. Herein, we have used mass spectrometry to study the oxidative metabolism of abacavir in EBV-transformed human B-cells. RNA and protein were isolated from the cells and subjected to transcriptomic and mass spectrometric analyses to identify the redox enzymes expressed. Low levels of isomeric abacavir carboxylic acids were detected in subcellular fractions of EBV-transformed human B-cells incubated with abacavir. Metabolite formation was time-dependent but was not reduced by an inhibitor of Class I alcohol dehydrogenases. Relatively high levels of mRNA were detected for several redox enzymes, including alcohol dehydrogenase 5 (Class III), aldehyde dehydrogenases (ALDH3A2, ALDH6A1, and ALDH9A1), CYP1B1, CYP2R1, CYP7B1, and hydroxysteroid dehydrogenase 10. Over 2600 proteins were identified by mass spectrometry. More than 1000 of these proteins exhibited catalytic activity, and 80 were oxido-reductases. This is the first proteomic inventory of enzymes in antigen presenting cells. However, neither of the hepatic alcohol dehydrogenases of Class I which metabolize abacavir in vitro was expressed at the protein level. Nevertheless the metabolic production of abacavir carboxylic acids by B-cell fractions implies abacavir-treated immune cells might be exposed to the drug's protein-reactive aldehyde metabolites in vivo.

  5. Wheat gliadin fractions and other cereal antigens reactive with antibodies in the sera of coeliac patients.

    PubMed Central

    Kieffer, M; Frazier, P J; Daniels, N W; Coombs, R R

    1982-01-01

    The mixed reverse solid phase passive antiglobulin haemadsorption test (MRSPAH) and the enzyme linked immunosorbant assay (ELISA) were found equally sensitive and fitted for the measurement of serum IgG antibodies against alcohol soluble gliadins. Using the ELISA method, three coeliac sera with elevated antibody titres against gliadins and two control sera with low titres were tested for IgG antibodies against the main groups of wheat proteins (acetic acid soluble glutenins, salt soluble albumins and globulins and alcohol soluble gliadins), eight fractions of gliadin and the alcohol soluble proteins of barley, rye, oat, maize and rice. As rice contained little alcohol soluble protein, a test against acid soluble rice proteins was included. In all three patient sera, titres higher than or equal to that for crude gliadin were found for wheat glutenin and for gliadin fractions seven and eight, both containing alpha gliadins. Similar high titres were found when these coeliac sera were tested against rye, barley and oat prolamines. Maize prolamines gave only low titres and no antibodies could be detected against rice proteins, in line with the tolerance of these latter two cereals by patients with coeliac disease. It would appear that sera from coeliac patients react with more than one antigenic fraction of protein in wheat and other cereals. Also sera from two normal persons appeared to have the same spectrum of reactivity against these cereal proteins as did the three sera from coeliac patients. The titres in normal sera were however much lower. PMID:7166001

  6. Immune response to different fractions of Taenia solium cyst fluid antigens in patients with neurocysticercosis.

    PubMed

    Amit, Prasad; Prasad, Kashi Nath; Kumar, Gupta Rakesh; Shweta, Tripathi; Sanjeev, Jha; Kumar, Paliwal Vimal; Mukesh, Tripathi

    2011-03-01

    The immunopathogenesis of neurocysticercosis (NCC) largely remains unknown. We analyzed the immune response to different fractions of Taenia solium cyst fluid antigens in patients with NCC. Lymphocytes were separated from 48 patients with NCC-related active epilepsy and 30 healthy controls. T. solium (isolated from pig muscles) antigens (crude lysate, CL; cyst wall, CW and cyst fluid, CF) at 20 μg/well concentrations were used to stimulate the cells in a lymphocyte transformation test (LTT). Only CF antigen stimulated cell proliferation significantly greater than control (p<0.001), hence cyst fluid antigens were further studied. The CF antigens were electro-blotted on nitrocellulose membrane (NC), cut at 0.5 cm distance and particulate antigens were prepared. A total of 12 fractions, designated F1 to F12 according to molecular weight were tested in-vitro for LTT. After 72 h of stimulation by the different fractions, Th1 (IL-1β, TNF-α, IL-2) and Th2 (IL-4, IL-10) cytokine responses were determined in culture supernatants by ELISA. Low molecular weight fractions F1 through F4 (Mol. wt.<25 kDa) were found to be potent inducers of cytokines. Fractions F1, F3 and F4 induced the production of Th1 (IL-1β, TNF-α, IL-2), whereas F2 induced the production of Th2 (IL-4 and IL-10) cytokine. The study shows that the low molecular weight fractions of CF antigens are immuno-dominant. Most of these fractions (F1, F3, F4) induce strong Th1 immune response except F2 which induces Th2 response. Further studies are needed to identify the different antigens present in these fractions to determine the molecules responsible for the immune response.

  7. Acid test: lipid antigens get into the groove.

    PubMed

    Kronenberg, Mitchell; Sullivan, Barbara A

    2008-06-01

    How do CD1 molecules load lipid antigens? In this issue of Immunity, Relloso et al. (2008) uncover how lysosomal pH targets amino acids in CD1b, causing it to open and attain a conformation more receptive to lipid antigens.

  8. Continuous flow magnetic cell fractionation based on antigen expression level.

    PubMed

    Schneider, Thomas; Moore, Lee R; Jing, Ying; Haam, Seungjoo; Williams, P Stephen; Fleischman, Aaron J; Roy, Shuvo; Chalmers, Jeffrey J; Zborowski, Maciej

    2006-07-31

    Cell separation is important in medical and biological research and plays an increasingly important role in clinical therapy and diagnostics, such as rare cancer cell detection in blood. The immunomagnetic labeling of cells with antibodies conjugated to magnetic nanospheres gives rise to a proportional relationship between the number of magnetic nanospheres attached to the cell and the cell surface marker number. This enables the potential fractionation of cell populations by magnetophoretic mobility (MM). We exploit this feature with our apparatus, the Dipole Magnet Flow Fractionator (DMFF), which consists of an isodynamic magnetic field, an orthogonally-oriented thin ribbon of cell suspension in continuous sheath flow, and ten outlet flows. From a sample containing a 1:1 mixture of immunomagnetically labeled (label+) and unlabeled (label-) cells, we achieved an increase in enrichment of the label+ cell fraction with increasing outlet numbers in the direction of the magnetic field gradient (up to 10-fold). The total recovery of the ten outlet fractions was 90.0+/-7.7%. The mean MM of label+ cells increased with increasing outlet number by up to a factor of 2.3. The postulated proportionality between the number of attached magnetic beads and the number of cell surface markers was validated by comparison of MM measured by cell tracking velocimetry (CTV) with cell florescence intensity measured by flow cytometry.

  9. Polarity based fractionation of fulvic acids.

    PubMed

    Li, Aimin; Hu, Jundong; Li, Wenhui; Zhang, Wei; Wang, Xuejun

    2009-11-01

    Fulvic acids from the soil of Peking University (PF) and a Nordic river (NF) were separated into well defined sub-fractions using sequential elution techniques based on eluent polarity. The chemical properties of the fractions including: PF1 and NF1 (eluted by 0.01 M HCl), PF2 and NF2 (eluted by 0.01 M HCl+20% methanol), PF3 and NF3 (eluted by 0.01 M HCl+40% methanol), and PF4 and NF4 (eluted by 100% methanol), were characterized using UV-Visible spectroscopy, elemental analysis and (13)C NMR. The results showed that the UV absorptions of the elution peaks at 280 nm (A280) increased from PF2 to PF4 and NF2 to NF4. No elution peaks were observed for PF1 and NF1. The carbon contents increased from 43.34% to 51.90% and 43.06% to 53.26% while the oxygen contents decreased from 46.39% to 36.76% and 49.76% to 40.03% for PF1-PF4 and NF1-NF4, respectively. As a polarity indicator, the (O+N)/C ratio for PF1-PF4 and NF1-NF4 decreased from 0.88 to 0.62 and 0.89 to 0.58, respectively. The aromatic carbon content increased from PF1 to PF4 and NF1 to NF4, suggesting an increase of the hydrophobicity of these fractions. The polarity was positively related to the ratio of UV absorption at 250 nm and 365 nm (E2/E3), and negatively related to the aromaticity. A high positive relationship between the aromaticity and E2/E3 of fulvic acid fractions was also obtained. The use of an eluent with a decreasing polarity allowed to providing simpler fractions of soil and aquatic fulvic acids.

  10. Serodiagnosis of fasciolosis by fast protein liquid chromatography-fractionated excretory/secretory antigens.

    PubMed

    Mokhtarian, Kobra; Akhlaghi, Lame; Meamar, Ahmad Reza; Razmjou, Elham; Manouchehri Naeini, Kourosh; Gholami, Samaneh; Najafi Samei, Masoomeh; Falak, Reza

    2016-08-01

    In several studies, different antigenic preparations and diverse immunological tests were applied for serodiagnosis of Fasciola hepatica infections. Most of these preparations showed cross-reactivity with proteins of other parasites. Application of purified antigens might reduce these cross-reactivities. Here, we used fast protein liquid chromatography (FPLC)-fractionated extracts of F. hepatica excretory/secretory antigens (E/S Ags) for serodiagnosis of human and sheep fasciolosis. To develop an improved diagnostic method, we fractionated F. hepatica E/S Ags by anion exchange chromatography on a Sepharose CL-6B column and then tested the serodiagnostic values of the fractions. We used sera from F. hepatica-infected human and sheep as positive controls. Sera from patients with hydatidosis and strongyloidiasis were used for cross-reactivity studies. Enzyme-linked immunosorbent assays (ELISA) of the second FPLC peak, containing 20, 25, and 70 kDa proteins, discriminated between F. hepatica-infected and uninfected human and sheep samples. Fractionation of F. hepatica E/S Ags by FPLC is a fast and reproducible way of obtaining antigens useful for serodiagnosis of human and sheep fasciolosis with acceptable sensitivity and specificity. Graphical abstract ᅟ.

  11. Copper isotope fractionation in acid mine drainage

    USGS Publications Warehouse

    Kimball, B.E.; Mathur, R.; Dohnalkova, A.C.; Wall, A.J.; Runkel, R.L.; Brantley, S.L.

    2009-01-01

    We measured the Cu isotopic composition of primary minerals and stream water affected by acid mine drainage in a mineralized watershed (Colorado, USA). The ??65Cu values (based on 65Cu/63Cu) of enargite (??65Cu = -0.01 ?? 0.10???; 2??) and chalcopyrite (??65Cu = 0.16 ?? 0.10???) are within the range of reported values for terrestrial primary Cu sulfides (-1??? < ??65Cu < 1???). These mineral samples show lower ??65Cu values than stream waters (1.38??? ??? ??65Cu ??? 1.69???). The average isotopic fractionation (??aq-min = ??65Cuaq - ??65Cumin, where the latter is measured on mineral samples from the field system), equals 1.43 ?? 0.14??? and 1.60 ?? 0.14??? for chalcopyrite and enargite, respectively. To interpret this field survey, we leached chalcopyrite and enargite in batch experiments and found that, as in the field, the leachate is enriched in 65Cu relative to chalcopyrite (1.37 ?? 0.14???) and enargite (0.98 ?? 0.14???) when microorganisms are absent. Leaching of minerals in the presence of Acidithiobacillus ferrooxidans results in smaller average fractionation in the opposite direction for chalcopyrite (??aq-mino = - 0.57 ?? 0.14 ???, where mino refers to the starting mineral) and no apparent fractionation for enargite (??aq-mino = 0.14 ?? 0.14 ???). Abiotic fractionation is attributed to preferential oxidation of 65Cu+ at the interface of the isotopically homogeneous mineral and the surface oxidized layer, followed by solubilization. When microorganisms are present, the abiotic fractionation is most likely not seen due to preferential association of 65Cuaq with A. ferrooxidans cells and related precipitates. In the biotic experiments, Cu was observed under TEM to occur in precipitates around bacteria and in intracellular polyphosphate granules. Thus, the values of ??65Cu in the field and laboratory systems are presumably determined by the balance of Cu released abiotically and Cu that interacts with cells and related precipitates. Such isotopic signatures

  12. Plasma lipid-bound sialic acid and carcinoembryonic antigen in cancer patients.

    PubMed

    Dnistrian, A M; Schwartz, M K

    1981-10-01

    We evaluated lipid-bound sialic acid as a "marker" in cancer patients and assessed the individual and combined value of lipid-bound sialic acid and carcinoembryonic antigen determinations in these patients. Plasma was sampled from 62 normal subjects and 125 cancer patients. Lipid-bound sialic acid was determined by the resorcinol method after total lipid extraction and isolation of the sialolipid fraction from plasma. Neither marker was increased in many breast cancer patients. Carcinoembryonic antigen was increased more commonly and to a greater degree in colon cancer patients and seems to be the preferred marker. Both markers were increased in lung cancer patients and their combined evaluation improved the rate of detection. Lipid-bound sialic acid was increased in more patients with leukemias, lymphomas, Hodgkin's disease, and melanomas, suggesting that it may be a useful biochemical marker in these types of cancer.

  13. Use of fractionated urinary filarial antigen in the diagnosis of human filariasis.

    PubMed

    Ramaprasad, P; Harinath, B C

    1995-02-01

    Fractionated urinary filarial antigen UFA C2 has shown high antigenic activity after absorption of urinary albumin present in the fraction. As little as 500 ag (10(-18) g) of albumin absorbed UFA C2, labelled as UFA C2-A, was found to be sufficient to detect filarial antibody. Stick enzyme immunoassay to assess the immunodiagnostic potential of UFA C2-A indicated filarial IgG antibody in 89% of microfilaraemic (mf) cases, 84% of clinical filariasis and 7% of endemic normals. UFA C2-A was found to be present in circulation in active as well as clinical infections as observed by inhibition assay using UFA C2-A penicillinase conjugate. Eighty-six per cent of mf, 50% of clinical cases and 6% of endemic normal subjects revealed parasite antigen to UFA C2-A on further serological analysis. None of the non-endemic normal sera showed the presence of filarial antibody/antigen to UFA C2-A. Furthermore, the test to determine phosphorylcholine (PC) bearing epitopes in UFA C2-A indicated no immunological reaction with anti-PC monoclonal antibody by avidin-biotin enzyme linked immunosorbent assay (ELISA). The highly sensitive and more easily obtainable non-PC urinary filarial antigen, UFA C2-A, is of great immunodiagnostic interest for lymphatic filariasis.

  14. Immunoaffinity fractionation of Schistosoma mansoni worm antigens using human antibodies and its application for serodiagnosis.

    PubMed Central

    Boctor, F N; Shaheen, H I

    1986-01-01

    A crude Schistosoma mansoni soluble worm antigen preparation (SWAP) was fractionated using an immunoaffinity column consisting of specific human anti-SWAP antibodies obtained from chronic S. mansoni-infected human sera and bound to CNBr-activated Sepharose 4B. The chromatographic separation resulted in three fractions: the unbound material (FW), and the eluted antigens with glycine-HCl (F1) and glycine-HCl-NaCl (F2). Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) showed that the purified antigens F1 and F2 consisted of several bands when stained with Coomassie blue and silver stain, with molecular weights between 20 X 10(3) and 200 X 10(3). The F1 and F2 fractions in addition to FW and SWAP were used in an enzyme-linked immunosorbent assay (ELISA) to measure antibody levels in sera from schistosomiasis patients. Each individual serum assessed with the purified F2 antigen gave 100% positivity and three to four times higher optical density in comparison to SWAP with only 88% positivity. No detectable cross-reactive antibodies against F2 were found when a limited number of sera from filariasis, fascioliasis and trichinellosis patients were screened. Furthermore, F2 was also used and found to be more sensitive generally in detecting anti-adult worm antibodies than SWAP in recently schistosomiasis-infected persons. Thus, F2 appears to be a highly sensitive and specific reagent for the serodiagnosis of schistosomiasis infection. Images Figure 3 Figure 7 PMID:3082749

  15. ANTIGENICITY OF POLYPEPTIDES (POLY ALPHA AMINO ACIDS)

    PubMed Central

    Pinchuck, Paul; Maurer, Paul H.

    1965-01-01

    The response of mice to synthetic linear polypeptides of known composition but random sequence has been studied. Neither Swiss mice nor a number of inbred strains could respond to copolymers of only 2 amino acids (G60L40, G60A40, G90T10). Upon introduction of as little as 4 mole per cent of a third amino acid, good immune responses were obtained, regardless of the nature of the third amino acid. The level of the immune response to a series of glu-lys-ala polymers increased with increasing alanine content of the polymer. PMID:5849232

  16. IMMUNODIAGNOSIS OF HUMAN STRONGYLOIDIASIS: USE OF SIX DIFFERENT ANTIGENIC FRACTIONS FROM Strongyloides venezuelensis PARASITIC FEMALES

    PubMed Central

    CORRAL, Marcelo Andreetta; de PAULA, Fabiana Martins; GOTTARDI, Maiara; MEISEL, Dirce Mary Correia Lima; CASTILHO, Vera Lucia Pagliusi; GONÇALVES, Elenice Messias do Nascimento; CHIEFFI, Pedro Paulo; GRYSCHEK, Ronaldo Cesar Borges

    2015-01-01

    SUMMARY The aim of this study was to evaluate six different antigenic fractions from Strongyloides venezuelensis parasitic females for the immunodiagnosis of human strongyloidiasis. Soluble and membrane fractions from S. venezuelensis parasitic females were prepared in phosphate-buffered saline (SSF and SMF, respectively), Tris-HCl (TSF and TMF, respectively), and an alkaline buffer (ASF and AMF, respectively). Serum samples obtained from patients with strongyloidiasis or, other parasitic diseases, and healthy individuals were analyzed by enzyme-linked immunosorbent assay (ELISA). Soluble fractions SSF, TSF, and ASF showed 85.0%, 75.0%, and 80.0% sensitivity and 93.1%, 93.1%, and 87.5% specificity, respectively. Membrane fractions SMF, TMF, and AMF showed 80.0%, 75.0%, and 85.0% sensitivity, and 95.8%, 90.3%, and 91.7% specificity, respectively. In conclusion, the present results suggest that the fractions obtained from parasitic females, especially the SSF and SMF, could be used as alternative antigen sources in the serodiagnosis of human strongyloidiasis. PMID:26603231

  17. Antigen-binding site protection during radiolabeling leads to a higher immunoreactive fraction

    SciTech Connect

    Van den Abbeele, A.D.; Aaronson, R.A.; Daher, S.; Taube, R.A.; Adelstein, S.J.; Kassis, A.I. )

    1991-01-01

    It is generally accepted that the immunointegrity of an antibody (Ab) depends on the preservation of its antigen-binding sites. Our goal was to radiolabel an antibody at several iodine:antibody molar ratios under conditions protecting its combining site and to compare its immunoreactive fraction (IRF) and electrophoretic mobility with those of the same antibody radiolabeled without protection. The data indicate that an antibody radiolabeled while its antigen-binding site is occupied by its antigen had the same IRF, regardless of the number of iodine atoms per antibody molecule. On the other hand, even at an I:Ab ratio of 1:1, the IRF of the same antibody radiolabeled without protection was lower than that of a protected one and decreased with increasing I:Ab ratios. In addition, the iodination of these Ab changes their electrophoretic mobility; however, when the Ab is labeled in the protected state, the degree of change is less. The binding of an antibody to its antigen prior to radiolabeling, therefore, enhances its immuno-integrity and prevents major conformational changes as reflected by electrophoresis.

  18. Enzymatic hydrolysis of heated whey: iron-binding ability of peptides and antigenic protein fractions.

    PubMed

    Kim, S B; Seo, I S; Khan, M A; Ki, K S; Lee, W S; Lee, H J; Shin, H S; Kim, H S

    2007-09-01

    This study evaluated the influence of various enzymes on the hydrolysis of whey protein concentrate (WPC) to reduce its antigenic fractions and to quantify the peptides having iron-binding ability in its hydrolysates. Heated (for 10 min at 100 degrees C) WPC (2% protein solution) was incubated with 2% each of Alcalase, Flavourzyme, papain, and trypsin for 30, 60, 90, 120, 150, 180, and 240 min at 50 degrees C. The highest hydrolysis of WPC was observed after 240 min of incubation with Alcalase (12.4%), followed by Flavourzyme (12.0%), trypsin (10.4%), and papain (8.53%). The nonprotein nitrogen contents of WPC hydrolysate followed the hydrolytic pattern of whey. The major antigenic fractions (beta-lactoglobulin) in WPC were degraded within 60 min of its incubation with Alcalase, Flavourzyme, or papain. Chromatograms of enzymatic hydrolysates of heated WPC also indicated complete degradation of beta-lactoglobulin, alpha-lactalbumin, and BSA. The highest iron solubility was noticed in hydrolysates derived with Alcalase (95%), followed by those produced with trypsin (90%), papain (87%), and Flavourzyme (81%). Eluted fraction 1 (F-1) and fraction 2 (F-2) were the respective peaks for the 0.25 and 0.5 M NaCl chromatographic step gradient for analysis of hydrolysates. Iron-binding ability was noticeably higher in F-1 than in F-2 of all hydrolysates of WPC. The highest iron contents in F-1 were observed in WPC hydrolysates derived with Alcalase (0.2 mg/kg), followed by hydrolysates derived with Flavourzyme (0.14 mg/kg), trypsin (0.14 mg/kg), and papain (0.08 mg/kg). Iron concentrations in the F-2 fraction of all enzymatic hydrolysates of WPC were low and ranged from 0.03 to 0.05 mg/kg. Fraction 1 may describe a new class of iron chelates based on the reaction of FeSO4 x 7 H2O with a mixture of peptides obtained by the enzymatic hydrolysis of WPC. The chromatogram of Alcalase F-1 indicated numerous small peaks of shorter wavelengths, which probably indicated a variety of

  19. Definition of glomerular antigens by monoclonal antibodies produced against a human glomerular membrane fraction.

    PubMed

    Neale, T J; Callus, M S; Donovan, L C; Baird, H

    1990-10-01

    Experimental animal models of glomerulonephritis (GN) produced by direct antibody binding to non-basement membrane glomerular capillary wall antigens do not to date have human parallels. To examine the potential for this form of humoral glomerular injury in man, we sought to define discrete human non-GBM glomerular antigenic targets using hybridoma technology. Mice were immunised intraperitoneally with 20-100 micrograms of a human glomerular membrane fraction (HGMF). Six fusions have yielded 12 stable reagents defined by positive glomerular indirect immunofluorescence (IF) and microELISA using HGMF as the screening antigen. Subclass analysis of ascitic McAbs indicated several IgG1, one IgG2b, and three IgM reagents. Distinctive IF patterns of reactivity with epithelial, endothelial or mesangial structures have been observed, with or without peritubular capillary, tubular basement membrane and vessel wall reactivity. Seven normal non-renal human organs and the kidneys of rat, rabbit and sheep have shown patterns characteristic of each individual McAb, restricted to human or with species cross reactivity. To partially characterise McAb-reactive antigens, detergent-solubilised renal cortex and collagenase-solubilised GBM (CS-GBM) extracts have been probed by immunoblot. A unique McAb 7-5Q, reactive with glomerular and tubular epithelial structures, binds major bands of approximately 107 KD and 93 KD in detergent solubilised cortex and a single band of similar size by immunoprecipitation (110 KD). 5-3A (a human-restricted linear-reacting McAb) binds bands of 20-200 KD (major band 58 KD) in CS-GBM. In conclusion, distinct species-restricted and more broadly disposed glomerular epitopes are definable in man by McAbs and are potential targets for humoral injury. Purification of these antigens will allow assay for circulating putative nephritogenic auto-antibody and potentially, McAbs may be useful in screening urine for evidence of occult structural renal disease.

  20. Adsorption characterizations of fulvic acid fractions onto kaolinite.

    PubMed

    Li, Aimin; Xu, Minjuan; Li, Wenhui; Wang, Xuejun; Dai, Jingyu

    2008-01-01

    Fulvic acids extracted from a typical rice-production region near Taihu Lake of China were fractionated into three fractions including F4.8, F7.0 and F11.0 (eluted by pH 4.8 buffer, pH 7.0 buffer and pH 11.0 buffer, respectively). Sorption of fulvic acid (FA) fractions onto kaolinite was studied by batch adsorption experiments, and characterizations of kaolinite before and after adsorption were investigated using scanning electron microscopy (SEM). Adsorption isotherms of kaolinite for three FA fractions fit well with the Langmuir adsorption model. The adsorption density of the three fractions was positively correlated with the ratio of the amount of the alkyl carbon to that of carboxyl and carbonyl carbon in FA fractions and followed an order of F11.0 > F7.0 > F4.8. Hydrophobic interaction was one of the control mechanisms for the sorption of FA fraction onto kaolinite. SEM images confirmed that compared to blank kaolinite samples, kaolinite samples coated by a FA fraction displayed an opener and more dispersed conformation resulting from the disruption of the floc structure in complex. Dispersion of kaolinite after adsorption was due to the repulsion between negatively charged FA-coated particles, which is closely related to the amount of FA fractions absorbed on kaolinite.

  1. Functional characterization of lymphocyte response to fractionated house dust mite antigens (Dermatophagoides pteronyssinus) in atopic and non-atopic individuals.

    PubMed Central

    Kimura, J Y; Ohta, N; Ishii, A; Nagano, T; Usui, M

    1990-01-01

    This study investigated the cellular response of human lymphocytes to Dermatophagoides pteronyssinus crude antigen and 14 molecular weight (MW) fractions. The cells were derived from atopic patients and healthy individuals who were skin test-positive or skin test-negative to mite antigen. When stimulated with crude antigen, the group of patients showed elevated proliferation and production of lymphokine in comparison with the healthy skin test-negative individuals (P less than 0.01). By stimulation with fractions, there was a remarkable variety in the responding patterns to each fraction. However, when expressed as a mean value, the patient group exhibited a sharp and high peak at 95,000 MW, which is different from IgE responses. In the other two groups, no apparent peak response was observed. Lymphokine production by fraction stimulation was studied in six distinct individuals. The most important fractions for inducing lymphokine production differed in each individual tested. Moreover, fractions which induced active lymphokine production were not necessarily the main targets of proliferative response in atopic patients. PMID:2379943

  2. Amino acid sequence and comparative antigenicity of chicken metallothionein.

    PubMed Central

    McCormick, C C; Fullmer, C S; Garvey, J S

    1988-01-01

    The complete amino acid sequence of metallothionein (MT) from chicken liver is reported. The primary structure was determined by automated sequence analysis of peptides produced by limited acid hydrolysis and by trypsin digestion. The comparative antigenicity of chicken MT was determined by radioimmunoassay using rabbit anti-rat MT polyclonal antibody. Chicken MT consists of 63 amino acids as compared to 61 found in MTs from mammals. One insertion (and two substitutions) occurs in the amino-terminal region, a region considered invariant among mammalian MTs. Eighteen of the 20 cysteines in chicken MT were aligned with cysteines from other mammalian sequences. Two cysteines near the carboxyl terminus are shifted by one residue due to the insertion of proline in that region. Overall, the chicken protein showed approximately equal to 68% sequence identity in a comparison with various mammalian MTs. The affinity of the polyclonal antibody for chicken MT was decreased by 2 orders of magnitude in comparison to that of a mammalian MT (rat MT isoforms). This reduced affinity is attributed to major substitutions in chicken MT in the regions of the principal determinants of mammalian MTs. Theoretical analysis of the primary structure predicted the secondary structure to consist of reverse turns and random coils with no stable beta or helix conformations. There is no evidence that chicken MT differs functionally from mammalian MTs. PMID:2448773

  3. MDR-TB Antibody Response (Western Blot) to Fractions of Isoniazid and Rifampicin Resistant Antigens of Mycobacterium tuberculosis.

    PubMed

    Hadizadeh Tasbiti, Alireza; Yari, Shamsi; Ghanei, Mostafa; Shokrgozar, Mohammad Ali; Bahrmand, Ahmadreza

    2015-12-01

    Drug-resistant TB poses a major threat to control of TB worldwide. Despite progress in the detection of Multidrug-resistant TB (MDR-TB) cases, a major diagnostic gap remains: 55% of reported TB patients estimated to have MDR-TB were not detected in 2013. MDR-TB antigens were conjugated to CNBr-activated Sepharose 4B. Specific polyclonal antibodies against MDR-TB Ags were prepared in rabbits using two boosted injections of the MDR-TB antigen. The antibodies were purified and treated with susceptible TB to remove any non-specific and cross-reactive antibodies. In the present study, comparative analysis of electrophoretic pattern of different antigens of INH/RIF-resistant TB were studied for identifying protein profiles. A RIF-resistant TB antigen was shown here to have different protein profiles from INH-resistant TB isolate. The results of Western blotting analysis showed that in the RIF- and INH-resistant antigenic fractions some bands of 14.4 and 45 kDa as immunogenic were common. Moreover, four bands of RIF-resistant TB antigen fractions (16, 19, 21, and 45 KDa) and one band of INH-resistant TB (about 26 KDa) were detected as diagnostic antigens. This study suggests that the Western blot is an accurate test to survey INH- and RIF-resistant TB antigens of M. tuberculosis infection. These findings indicate that MDR-TB diagnosis (based on Ag detection) could be useful in the identification of disease stages that precede symptomatic and microbiologically positive TB, such as subclinical and incipient TB.

  4. A polysaccharide fraction from medicinal herb Prunella vulgaris downregulates the expression of herpes simplex virus antigen in Vero cells.

    PubMed

    Chiu, Lawrence Chi-Ming; Zhu, Wen; Ooi, Vincent Eng-Choon

    2004-07-01

    Herpes simplex viruses (HSV) are pathogenic. With the emergence of drug-resistant strains of HSV, new antiviral agents, especially those with different modes of action, are urgently needed. Prunella vulgaris L. (Labiatae), a perennial plant commonly found in China and Europe, has long been used as a folk medicine to cure ailments. In this study, a polysaccharide fraction was prepared from Prunella vulgaris (PPV), and its effects on the expressions of HSV-1 and HSV-2 antigens in their host Vero cells were investigated with flow cytometry. The HSV antigen increased time-dependently in the infected cells, and PPV reduced its expression. The effective concentrations of PPV with 50% reductions of the HSV-1 and HSV-2 antigens were 20.6 and 20.1 microg/ml, respectively. The novelty of PPV is that it also reduces the antigen expression of acyclovir-resistant strain of HSV-1. After incubations with 25-100 microg/ml of PPV the HSV antigen-positive cells were reduced by 24.8-92.6%, respectively, showing that this polysaccharide fraction has a different mode of anti-HSV action from acyclovir. Results from this study show that PPV is effective against both the HSV-1 and HSV-2 infections, and flow cytometry offers a quantitative and highly reproducible anti-HSV drug-susceptibility assay.

  5. Rapid T cell–based identification of human tumor tissue antigens by automated two-dimensional protein fractionation

    PubMed Central

    Beckhove, Philipp; Warta, Rolf; Lemke, Britt; Stoycheva, Diana; Momburg, Frank; Schnölzer, Martina; Warnken, Uwe; Schmitz-Winnenthal, Hubertus; Ahmadi, Rezvan; Dyckhoff, Gerhard; Bucur, Mariana; Jünger, Simone; Schueler, Thomas; Lennerz, Volker; Woelfel, Thomas; Unterberg, Andreas; Herold-Mende, Christel

    2010-01-01

    Identifying the antigens that have the potential to trigger endogenous antitumor responses in an individual cancer patient is likely to enhance the efficacy of cancer immunotherapy, but current methodologies do not efficiently identify such antigens. This study describes what we believe to be a new method of comprehensively identifying candidate tissue antigens that spontaneously cause T cell responses in disease situations. We used the newly developed automated, two-dimensional chromatography system PF2D to fractionate the proteome of human tumor tissues and tested protein fractions for recognition by preexisting tumor-specific CD4+ Th cells and CTLs. Applying this method using mice transgenic for a TCR that recognizes an OVA peptide presented by MHC class I, we demonstrated efficient separation, processing, and cross-presentation to CD8+ T cells by DCs of OVA expressed by the OVA-transfected mouse lymphoma RMA-OVA. Applying this method to human tumor tissues, we identified MUC1 and EGFR as tumor-associated antigens selectively recognized by T cells in patients with head and neck cancer. Finally, in an exemplary patient with a malignant brain tumor, we detected CD4+ and CD8+ T cell responses against two novel antigens, transthyretin and calgranulin B/S100A9, which were expressed in tumor and endothelial cells. The immunogenicity of these antigens was confirmed in 4 of 10 other brain tumor patients. This fast and inexpensive method therefore appears suitable for identifying candidate T cell antigens in various disease situations, such as autoimmune and malignant diseases, without being restricted to expression by a certain cell type or HLA allele. PMID:20458140

  6. Serodiagnosis of human neurocysticercosis using antigenic components of Taenia solium metacestodes derived from the unbound fraction from jacalin affinity chromatography.

    PubMed

    Machado, Gleyce Alves; Oliveira, Heliana Batista de; Gennari-Cardoso, Margareth Leitão; Mineo, José Roberto; Costa-Cruz, Julia Maria

    2013-05-01

    The aim of the present study was to analyse Taenia solium metacestode antigens that were derived from the unbound fraction of jacalin affinity chromatography and subsequent tert-octylphenoxy poly (oxyethylene) ethanol Triton X-114 (TX-114) partitioning in the diagnosis of human neurocysticercosis (NCC). Immunoassays were designed to detect T. solium-specific IgG antibodies by ELISA and immunoblot. Serum samples were collected from 132 individuals who were categorised as follows: 40 had NCC, 62 presented Taenia spp or other parasitic diseases and 30 were healthy individuals. The jacalin-unbound (J unbound ) fraction presented higher sensitivity and specificity rates than the jacalin-bound fraction and only this fraction was subjected to subsequent TX-114 partitioning, resulting in detergent (DJ unbound ) and aqueous (AJ unbound ) fractions. The ELISA sensitivity and specificity were 85% and 84.8% for J unbound , 92.5% and 93.5% for DJ unbound and 82.5% and 82.6% for AJ unbound . By immunoblot, the DJ unbound fraction showed 100% sensitivity and specificity and only serum samples from patients with NCC recognised the 50-70 kDa T. solium-specific components. We conclude that the DJ unbound fraction can serve as a useful tool for the differential immunodiagnosis of NCC by immunoblot.

  7. Determining the Δ47 acid fractionation in dolomites

    NASA Astrophysics Data System (ADS)

    Murray, Sean T.; Arienzo, Monica M.; Swart, Peter K.

    2016-02-01

    This paper reports the fractionation of Δ47 during the digestion of dolomite in phosphoric acid between 25 °C and 90 °C using five different samples, including three of Pliocene age from the Bahamas, one from the Jurassic in the Middle East, and one obtained from the National Institute of Standards (NIST 88b). The composition of the dolomites analyzed varied from Ca0.56Mg0.44CO3 to Ca0.50Mg0.50CO3. Fractionation values were also compared between the common acid bath and sealed vessel techniques at various temperatures. No statistically significant differences were observed either between these two methods or as a function of the dolomite's stoichiometry. These data produce a difference in fractionation of 0.153 ± 0.011‰ for dolomite samples digested at 90 °C compared to those reacted at 25 °C, a value higher and statistically different to previously published values. Utilization of this value in a study of dolomites from a core drilled on the island of San Salvador in the Bahamas yielded temperatures and δ18Ofluid values which agree with previous interpretations on the formation of these dolomites. Application of this value to other published studies produces lower estimates of temperature and δ18Ofluid values for the dolomitization process that are more consistent with the geologic models suggested for these studies.

  8. Homogeneous electrochemical detection of hippuric acid in urine based on the osmium-antigen conjugate.

    PubMed

    Jeon, Won-Yong; Choi, Young-Bong; Kim, Hyug-Han

    2013-07-22

    A homogeneous electrochemical immunoassay is based on the interaction of osmium-antigen conjugate with its antibody. The novelty presented herein is the direct conjugation of the osmium complex to a small antigen and the application of the quantitative analysis of the antigen and its antibody as the electrical signal for homogeneous immunoassay. The small antigen chosen is hippuric acid (HA), a major urinary metabolite in toluene-exposed humans. As a redox mediator, [Os(4,4'-dimethoxy-2,2'-bipyridine)2(4-aminomethylpyridine-HA)Cl](+/2+) (Os-HA antigen) has been synthesized and characterized on screen-printed carbon electrodes. The synthesized Os-HA antigen shows reversible redox peaks at E(½)=0.056 V versus Ag/AgCl. The homogeneous competitive immunoassay relies on the interaction between Os-HA antigen conjugate and free antigen to its antibody, which can generate electrical signals linearly proportional to the free antigen monitored by cyclic voltammetry and differential pulse voltammetry in the range of 10 μg mL(-1) to 5.12 mg mL(-1). The cutoff concentration of HA in urine samples is 2.0 mg mL(-1), so the method can be used to develop a HA immunosensor. Moreover, the proposed homogeneous electrochemical immunoassay method can be applied to detect low concentrations of small antigens found in the healthcare area.

  9. [Synthesis and identification of penicillic-acid antigens from Penicillium cyclopium].

    PubMed

    Lei, Hongyu; Yuan, Hui; Wu, Jing; Yuan, Liyun; Wen, Lixin; Ni, Hengjia

    2008-05-01

    To establish a new immune assay for Penicillic Acid (PA) from Penicillium cyclopium, we studied the synthesis of conjugated complete antigens for penicillic acid. PA was conjugated to bovine serum album (BSA) and ovalbumin (OVA) by 1-ethyl-3-(3-dimethyl-aminopropyl) carbodiimide hydrochloride (EDC). The artificial antigens PA-BSA and PA-OVA were identified by ultraviolet spectrometric scanning, SDS-PAGE and immunization. Results showed that the absorption peak of conjugation were different from that of the carrier protein alone and of the PA. The conjugated ratio of PA and BSA was 23.2:1 and that of PA and OVA was 10.4:1. Balb/c mice were immunized by the artificial antigen of PA-BSA, with PA-OVA as coating antigen. The average titer of antiserums was more than 12 800 by indirect ELISA. The obtained antigens offered a basis for developing immunoassay method.

  10. Partial purification of fatty-acid binding protein by ammonium sulphate fractionation.

    PubMed

    Avanzati, B; Catalá, A

    1983-07-01

    By fractionation of rat liver cytosol with 70% saturation ammonium sulphate, a soluble fraction showing high affinity for oleic acid was obtained. The binding of oleic acid to this fraction was inhibited by flavaspidic acid. The molecular weight of the main protein present in this fraction was 12 000 as determined by SDS-poly-acrylamide-gel electrophoresis. This soluble fraction stimulated the transfer of oleic acid from microsomes to phosphatidylcholine liposomes as demonstrated by a transfer assay in vitro. The behaviour of this fraction is similar to that described for fatty-acid binding protein.

  11. Serodiagnosis of human neurocysticercosis using antigenic components of Taenia solium metacestodes derived from the unbound fraction from jacalin affinity chromatography

    PubMed Central

    Machado, Gleyce Alves; de Oliveira, Heliana Batista; Gennari-Cardoso, Margareth Leitão; Mineo, José Roberto; Costa-Cruz, Julia Maria

    2013-01-01

    The aim of the present study was to analyse Taenia solium metacestode antigens that were derived from the unbound fraction of jacalin affinity chromatography and subsequent tert-octylphenoxy poly (oxyethylene) ethanol Triton X-114 (TX-114) partitioning in the diagnosis of human neurocysticercosis (NCC). Immunoassays were designed to detect T. solium-specific IgG antibodies by ELISA and immunoblot. Serum samples were collected from 132 individuals who were categorised as follows: 40 had NCC, 62 presented Taenia spp or other parasitic diseases and 30 were healthy individuals. The jacalin-unbound (Junbound) fraction presented higher sensitivity and specificity rates than the jacalin-bound fraction and only this fraction was subjected to subsequent TX-114 partitioning, resulting in detergent (DJunbound) and aqueous (AJunbound) fractions. The ELISA sensitivity and specificity were 85% and 84.8% for Junbound, 92.5% and 93.5% for DJunboundand 82.5% and 82.6% for AJunbound. By immunoblot, the DJunboundfraction showed 100% sensitivity and specificity and only serum samples from patients with NCC recognised the 50-70 kDa T. solium-specific components. We conclude that the DJunboundfraction can serve as a useful tool for the differential immunodiagnosis of NCC by immunoblot. PMID:23778661

  12. Detection of Goodpasture antigen in fractions prepared from collagenase digests of human glomerular basement membrane.

    PubMed Central

    Fish, A J; Lockwood, M C; Wong, M; Price, R G

    1984-01-01

    Preparations of human glomerular basement membrane (GBM) were digested with collagenase, and a Goodpasture (GP) antigen rich pool from gel filtration column runs was identified by antibody inhibition radioimmunoassay. The components of the GP antigen pool were separated on polyacrylamide gels, and transferred to nitrocellulose sheets by the 'western' blotting technique. The blots were separately reacted with thirteen GP sera as primary antibody, followed by peroxidase labelled goat anti-human IgG and revealed 45-50K (two bands) and 25-28K (one-three bands) components. No corresponding reactivity was observed using convalescent GP sera or other control sera (normal human serum, rapidly progressive glomerulonephritis with or without pulmonary haemorrhage, and lupus erythematosus) as primary antibody. Images Fig. 3 PMID:6319059

  13. Role of polyadenylic acid in a deoxyribonucleic acid-membrane fraction extracted from pneumococci.

    PubMed Central

    Firshein, W; Meyer, B; Epner, E; Viggiani, J

    1976-01-01

    After the addition of radioactive polyadenylic acid to cell suspensions of pneumocci, part of the radioactivity becomes associated with a deoxyribonucleic acid (DNA)-membrane fraction extracted from the cells. A variety of techniques show that a portion of this associated radioactivity may represent oligoadenylates complexed to DNA, probaby as part of a ribonucleic acid (RNA) component. Polyadenylic acid, which had previously been shown to enhance DNA synthesis in cell suspensions (Firshein and Benson, 1968), also enhances the extent of DNA synthesis by the DNA-membrane fraction in vitro under specific conditions of concentration and conformation. The mechanism of action of this enhancement may be related to the ability of oligoadenylates to increase the number of initiation sites for DNA replication by stimulating the production of an RNA primer, thus providing additional 3'-OH groups with which DNA polymerase can react. PMID:6428

  14. Distribution and Characterization of Antigens Found in Subcellular Fractions of African Trypanosomes.

    DTIC Science & Technology

    1985-09-01

    affinity Ca-ATPase which we have described (2). Less specific acid phosphatases were also considered as potential surface membrane 3 markers since... acid phosphatase (0-glycerophosphate), arylamidase; promito- chondria -- DCPIP linked c-glycerophosphate dehydrogenase; lysosome -- proteinase...glucose-6- phosphatase , that were present in a I. brucei surface membrane preparation (22) are almost certainly due to the tartrate resistant acid

  15. Prophylactic efficacy of high-molecular-weight antigenic fractions of a recent clinical isolate of Leishmania donovani against visceral leishmaniasis.

    PubMed

    Tripathi, P; Gupta, S K; Sinha, S; Sundar, S; Dube, A; Naik, S

    2008-11-01

    T-cell mediated immune responses are key determinants to the natural course of infection caused by intracellular parasites such as Leishmania. Thus, T-cell activating proteins of these microbes continue to generate active interest particularly in view of their possible role in the design and development of newer and more effective vaccines. We have recently reported the presence of T-cell immunostimulatory antigens with the high-molecular-weight (MW) fractions (134-64.2 kDa) of whole Leishmania donovani antigen (strain 2001), which stimulated variable amounts of IFN-gamma, IL-12 and IL-10 in exposed immune individuals. The present study was undertaken to further evaluate these high-MW antigenic fractions (MW range >100-60 kDa) for potential protective efficacy. The high-MW region of the parasite was resolved into five antigenic fractions (Prep A-E) using continuous elution gel electrophoresis. Prior to in vivo protection studies in hamsters, these fractions were used to evaluate in vitro cellular responses in eight Leishmania-exposed individuals and treated cured hamsters. The protective efficacy of prep (A + B), C, D and E in combination with BCG was evaluated in inbred hamsters using standard immunization protocol. Proliferative responses were seen in all eight of eight exposed individuals to prep D [median stimulation index (SI): 5.2 (range 3.9-7.1)] and E [median SI: 5.6 (range 4.4-8.2)], five of eight individuals to prep B and prep C and three of eight to prep A [median SI: 0.2 (range 0.1-7.2)]. The median proliferative responses to prep D and prep E were significantly higher than to fraction prep A; (P < 0.05) but not to prep B and prep C. However, prep A-E induced equivalent levels of IFN-gamma, IL-10 and IL-12 cytokines. Fractions D and E also exhibited marked parasite inhibition in spleen (52.5% and 73.7%) and liver (65% and 80.2%) as compared with prep (A + B) (23% in spleen and 24% in liver) and prep C (38% in spleen and 24% in liver). Prep D and prep E

  16. Molecular size and amino acid composition of H-2d antigen solubilized in Nonidet P-40.

    PubMed

    Rossowski, W; Kloczewiak, M; Radzikowski, C; Strzadala, L

    1976-01-01

    H-2d antigenic material solubilized by the detergent Nonidet P-40 from L-1210 mouse leukemia cells was isolated by gel filtration on Bio-Gel P-100. A single peak eluted in the void volume consisted of about 90% protein, 8% hexose and traces of sialic acids. In sedimentation velocity runs, the antigen sedimented as a single peak of 3-1 S. Molecular weight determined by sedimentation equilibrium as well as calculated from amino acid composition was found to be in the range of 53,000 daltons and approx. 45,000-51,000 when calculated from sodium dodecyl sulfate polyacrylamide gel electrophoresis. Secondary structure of H-2d glycoprotein was predicted from the amino acid composition. For NP-40-solubilized H-2d antigen, about 34% of helix, 13% beta sheet and 41% turns was found.

  17. Distribution and Characterization of Antigens Found in Subcellular Fractions of African Trypanosomes.

    DTIC Science & Technology

    1982-08-01

    measured for relative fluorescence. These details are ndicated in Fig. A. ii. Lecting affinity chromatography. Either lentil lectin seph- arose 4B or...Zwaal, R.F.A. 1976. The use of phospholipases in the determination of asymmetric phospholipid distribution in membranes. In Methods in Membrane Biology 7...research. In Methods in Membrane Biology , Vol. 8 (Korn, E.D., ed.), pp. 1-50. 26 Enzyme Membrane Fraction Phosphatase SM FPM -glycerophosphate 10 15

  18. Synthetic oligonucleotide antigens modified with locked nucleic acids detect disease specific antibodies

    NASA Astrophysics Data System (ADS)

    Samuelsen, Simone V.; Solov’Yov, Ilia A.; Balboni, Imelda M.; Mellins, Elizabeth; Nielsen, Christoffer Tandrup; Heegaard, Niels H. H.; Astakhova, Kira

    2016-10-01

    New techniques to detect and quantify antibodies to nucleic acids would provide a significant advance over current methods, which often lack specificity. We investigate the potential of novel antigens containing locked nucleic acids (LNAs) as targets for antibodies. Particularly, employing molecular dynamics we predict optimal nucleotide composition for targeting DNA-binding antibodies. As a proof of concept, we address a problem of detecting anti-DNA antibodies that are characteristic of systemic lupus erythematosus, a chronic autoimmune disease with multiple manifestations. We test the best oligonucleotide binders in surface plasmon resonance studies to analyze binding and kinetic aspects of interactions between antigens and target DNA. These DNA and LNA/DNA sequences showed improved binding in enzyme-linked immunosorbent assay using human samples of pediatric lupus patients. Our results suggest that the novel method is a promising tool to create antigens for research and point-of-care monitoring of anti-DNA antibodies.

  19. Synthetic oligonucleotide antigens modified with locked nucleic acids detect disease specific antibodies

    PubMed Central

    Samuelsen, Simone V.; Solov’yov, Ilia A.; Balboni, Imelda M.; Mellins, Elizabeth; Nielsen, Christoffer Tandrup; Heegaard, Niels H. H.; Astakhova, Kira

    2016-01-01

    New techniques to detect and quantify antibodies to nucleic acids would provide a significant advance over current methods, which often lack specificity. We investigate the potential of novel antigens containing locked nucleic acids (LNAs) as targets for antibodies. Particularly, employing molecular dynamics we predict optimal nucleotide composition for targeting DNA-binding antibodies. As a proof of concept, we address a problem of detecting anti-DNA antibodies that are characteristic of systemic lupus erythematosus, a chronic autoimmune disease with multiple manifestations. We test the best oligonucleotide binders in surface plasmon resonance studies to analyze binding and kinetic aspects of interactions between antigens and target DNA. These DNA and LNA/DNA sequences showed improved binding in enzyme-linked immunosorbent assay using human samples of pediatric lupus patients. Our results suggest that the novel method is a promising tool to create antigens for research and point-of-care monitoring of anti-DNA antibodies. PMID:27775006

  20. Chemical composition of phase I Coxiella burnetii soluble antigen prepared by trichloroacetic acid extraction.

    PubMed

    Lukácová, M; Brezina, R; Schramek, S; Pastorek, J

    1989-01-01

    Optimal conditions of extraction (time and temperature) by trichloroacetic acid of soluble antigen from phase I Coxiella burnetii (TCAE), possessing protective properties and used as a chemovaccine against Q fever in men, were studied. Extracts prepared under various conditions were analysed for their polysaccharide, protein and phosphorus contents. Forty-five min of extraction at 0 degrees C were sufficient to obtain a soluble antigen reacting in immunodiffusion with hyperimmune rabbit antiserum. The polysaccharide contents decreased with prolonged extraction at 0 degrees C. At higher extraction temperatures (37 and 100 degrees C), the polysaccharide contents increased while that of proteins decreased. TCAE prepared at 100 degrees C gave no positive immunodiffusion reaction.

  1. Fractionation of humic acids upon adsorption on montmorillonite and palygorskite

    NASA Astrophysics Data System (ADS)

    Alekseeva, T. V.; Zolotareva, B. N.

    2013-06-01

    The adsorption of three humic acid (HA) preparations by clays—montmorillonite (Wyoming, USA) and palygorskite (Kolomenskoe district, Moscow oblast)—has been studied. The HA preparations were isolated from samples of the humus-accumulative horizons of a leached chernozem (Voronezh) and a chestnut soil (Volgograd), and a commercial preparation of sodium humate (Aldrich) was also used. The solid-state 13C NMR spectroscopy and IR spectroscopy revealed the selective adsorption of structural HA fragments (alkyls, O-alkyls (carbohydrates), and acetal groups) on these minerals. As a result, the aromaticity of the organic matter (OM) in the organic-mineral complexes (OMCs) and the degree of its humification have been found to be lower compared to the original HA preparations. The fractionation of HAs is controlled by the properties of the mineral surfaces. The predominant enrichment of OMCs with alkyls has been observed for montmorillonite, as well as an enrichment with O-alkyls (carbohydrates) for palygorskite. A decrease in the C : N ratio has been noted in the elemental composition of the OM in complexes, which reflected its more aromatic nature and (or) predominant sorption of N-containing structural components of HA molecules. The adsorption of HA preparations by montmorillonite predominantly occurs on the external surface of mineral particles, and the interaction of nonpolar alkyl groups of HAs with this mineral belongs to weak (van der Waals, hydrophobic) interactions. The adsorption of HA preparations by palygorskite is at least partly of chemical nature: Si-OH groups of minerals are involved in the adsorption process. The formation of strong bonds between the OM and palygorskite explains the long-term (over 300 million years) retention of fossil fulvate-type OM in its complex with palygorskite, which we revealed previously.

  2. Analysis of Known Bacterial Protein Vaccine Antigens Reveals Biased Physical Properties and Amino Acid Composition

    PubMed Central

    Mayers, Carl; Rowe, Sonya; Miller, Julie; Lingard, Bryan; Hayward, Sarah; Titball, Richard W.

    2003-01-01

    Many vaccines have been developed from live attenuated forms of bacterial pathogens or from killed bacterial cells. However, an increased awareness of the potential for transient side-effects following vaccination has prompted an increased emphasis on the use of sub-unit vaccines, rather than those based on whole bacterial cells. The identification of vaccine sub-units is often a lengthy process and bioinformatics approaches have recently been used to identify candidate protein vaccine antigens. Such methods ultimately offer the promise of a more rapid advance towards preclinical studies with vaccines. We have compared the properties of known bacterial vaccine antigens against randomly selected proteins and identified differences in the make-up of these two groups. A computer algorithm that exploits these differences allows the identification of potential vaccine antigen candidates from pathogenic bacteria on the basis of their amino acid composition, a property inherently associated with sub-cellular location. PMID:18629010

  3. Role of deoxyribonucleic acid ligase in a doxyribonucleic acid membrane fraction extracted from pneumococci.

    PubMed Central

    Greene, M; Firshein, W

    1976-01-01

    Deoxyribonucleic acid (DNA) ligase has been detected in a DNA membrane fraction extracted from Pneumococcus. The specific activity of the enzyme in this fraction is 10-fold greater than in the remaining cell extract. It remains firmly bound (with other enzymes) to the complex after a purification procedure in which a considerable percentage of the macromolecules are dissociated. The ligase acts in two ways in the DNA membrane fraction in vitro. One, it catalyzes the linkage of small-molecular-weight pieces of newly synthesized DNA into heavier-molecular-weight DNA strands as shown by others (M Gellert, 1976; R. Okazaki, A. Sugino, S. Hirose, T. Okazaki, Y. Imae, R. Kainuma-Kuroda, T. Ogawa, M. Arisawa, and Y. Kurosowa, 1973; B. Olivera and I. Lehman, 14; and A. Sugino, S. Hirose, and R. Okazaki, 1972) and, two, it protects DNA from degradation by deoxyribonucleases. This latter effect is due to a competition between the ability of the nucleases to degrade DNA and the ability of DNA ligase to seal the nicks produced by these degradative enzymes. The ligase acts cooperatively with other enzymes in the DNA membrane fraction to synthesize DNA. PMID:4433

  4. Antigenic fractions from Taenia crassiceps metacestodes obtained by hydrophobicity for the immunodiagnosis of active and inactive forms of neurocysticercosis in human cerebrospinal fluid samples.

    PubMed

    da Silva, Gabriela B; Nunes, Daniela S; de Sousa, José Eduardo N; Gonçalves-Pires, Maria do R F; Levenhagen, Marcelo A; Costa-Cruz, Julia M

    2017-04-01

    This study aimed to evaluate the total extract of Taenia crassiceps metacestodes (TC) and its antigenic fractions obtained by Triton X-114 fractionation techniques, such as detergent (DC) and aqueous (AC), in the immunodiagnosis of human neurocysticercosis (NCC). Cerebrospinal fluid samples were divided into two groups: Group 1 (n=40), which was further divided into active (n=20) and inactive (n=20) NCC, and Group 2 (control group), which comprised 39 CSF samples from patients who had another neurological disorder, were suffering from other infectious diseases of the brain or had other parasitic infections. The total extracts and antigenic fractions were tested by enzyme-linked immunosorbent assay (ELISA) to detect human IgG anti-Taenia solium. T. crassiceps fractions (DC and AC) showed the same value of sensitivity (Se), 100%, for active and inactive NCC and a specificity (Sp) of 97.4%. The DS fraction obtained from T. solium showed 100% Se for active NCC, 95% Se for inactive NCC and a 92.3% Sp. The AS fraction obtained from T. solium showed 100% Se for both active and inactive NCC and a 94.9% Sp. There was a positive correlation between the total saline extract of T. crassiceps (TC) and T. solium (TS) and their fractions (DC, AC, DS and AS). Positive predictive value, negative predictive value, diagnostic efficiency and Youden index were calculated. In conclusion, these results demonstrated that detergent and aqueous fractions obtained from T. crassiceps metacestodes are important sources of specific antigens and are efficient for immunodiagnosis of active and inactive NCC.

  5. Expression patterns of proliferating cell nuclear antigen in trichloroacetic acid peeled skin.

    PubMed

    Yamamoto, Yuki; Uede, Koji; Yonei, Nozomi; Furukawa, Fukumi

    2007-02-01

    This study was designed to investigate whether trichloroacetic acid (TCA) peeling induces cellular proliferation in human skin using an immunohistochemical method. A 40% TCA peel resulted in a greater number of proliferating cell nuclear antigen (PCNA)-immunopositive cells in the whole epidermis as compared with 60% TCA or phenol peels. This finding suggests that long-term and frequent TCA peelings of low concentration would require special attention for unexpected cutaneous lesions such as skin tumors.

  6. Concentration and fractionation of hydrophobic organic acid constituents from natural waters by liquid chromatography

    USGS Publications Warehouse

    Thurman, E.M.; Malcolm, R.L.

    1979-01-01

    A scheme is presented which used adsorption chromatography with pH gradient elution and size-exclusion chromatography to concentrate and separate hydrophobic organic acids from water. A review of chromatographic processes involved in the flow scheme is also presented. Organic analytes which appear in each aqueous fraction are quantified by dissolved organic carbon analysis. Hydrophobic organic acids in a water sample are concentrated on a porous acrylic resin. These acids usually constitute approximately 30-50 percent of the dissolved organic carbon in an unpolluted water sample and are eluted with an aqueous eluent (dilute base). The concentrate is then passed through a column of polyacryloylmorpholine gel, which separates the acids into high- and low-molecular-weight fractions. The high- and low-molecular-weight eluates are reconcentrated by adsorption chromatography, then are eluted with a pH gradient into strong acids (predominately carboxylic acids) and weak acids (predominately phenolic compounds). For standard compounds and samples of unpolluted waters, the scheme fractionates humic substances into strong and weak acid fractions that are separated from the low molecular weight acids. A new method utilizing conductivity is also presented to estimate the acidic components in the methanol fraction.

  7. Amino acid composition of Lagenaria siceraria seed flour and protein fractions.

    PubMed

    Ogunbusola, Moriyike Esther; Fagbemi, Tayo Nathaniel; Osundahunsi, Oluwatooyin Faramade

    2010-12-01

    Defatted seed flours of Lagenaria siceraria (calabash and bottle gourd) were fractionated into their major protein fractions. The amino acid composition of seed flours and their protein fractions were determined and the protein quality was evaluated. Glutamic acid (139-168 mg/g protein) was the most abundant amino acid followed by aspartic acid (89.0-116 mg/g protein) in both the seed flours and their protein fractions. The total essential amino acid ranged from 45.8 to 51.5%. The predicted protein efficiency ratio and the predicted biological value ranged from 2.4 to 2.9 and 8.7 to 44.0, respectively. Lysine and sulphur amino acids were mostly concentrated in the globulin fractions. The first and second limiting amino acids in seed flours and protein fractions were methionine and valine or threonine. The seed flours contained adequate essential amino acids required by growing school children and adults. The seed has potential as protein supplement in cereal based complementary diets or in the replacement of animal proteins in conventional foods.

  8. Comparison of aqueous ammonia and dilute acid pretreatment of bamboo fractions: Structure properties and enzymatic hydrolysis.

    PubMed

    Xin, Donglin; Yang, Zhong; Liu, Feng; Xu, Xueru; Zhang, Junhua

    2015-01-01

    The effect of two pretreatments methods, aqueous ammonia (SAA) and dilute acid (DA), on the chemical compositions, cellulose crystallinity, morphologic change, and enzymatic hydrolysis of bamboo fractions (bamboo yellow, timber, green, and knot) was compared. Bamboo fractions with SAA pretreatment had better hydrolysability than those with DA pretreatment. High crystallinity index resulted in low hydrolysis yield in the conversion of SAA pretreated bamboo fractions, not DA pretreated fractions. The increase of cellulase loading had modestly positive effect in the hydrolysis of both SAA and DA pretreated bamboo fractions, while supplement of xylanase significantly increased the hydrolysis of the pretreated bamboo fractions, especially after SAA pretreatment. The results indicated that SAA pretreatment was more effective than DA pretreatment in conversion of bamboo fractions, and supplementation of xylanase was necessary in effective conversion of the SAA pretreated fractions into fermentable sugars.

  9. Closed-system behaviour of the intra-crystalline fraction of amino acids in mollusc shells

    PubMed Central

    Penkman, K.E.H.; Kaufman, D.S.; Maddy, D.; Collins, M.J.

    2008-01-01

    When mollusc shells are analysed conventionally for amino acid geochronology, the entire population of amino acids is included, both inter- and intra-crystalline. This study investigates the utility of removing the amino acids that are most susceptible to environmental effects by isolating the fraction of amino acids encapsulated within mineral crystals of mollusc shells (intra-crystalline fraction). Bleaching, heating and leaching (diffusive loss) experiments were undertaken on modern and fossil Corbicula fluminalis, Margaritifera falcata, Bithynia tentaculata and Valvata piscinalis shells. Exposure of powdered mollusc shells to concentrated NaOCl for 48 h effectively reduced the amino acid content of the four taxa to a residual level, assumed to represent the intra-crystalline fraction. When heated in water at 140 °C for 24 h, only 1% of amino acids were leached from the intra-crystalline fraction of modern shells compared with 40% from whole shell. Free amino acids were more effectively retained in the intra-crystalline fraction, comprising 55% (compared with 18%) of the whole shell after 24 h at 140 °C. For fossil gastropods, the inter-shell variability in D/L values for the intra-crystalline fraction of a single-age population was reduced by 50% compared with conventionally analysed shells. In contrast, analysis of the intra-crystalline fraction of C. fluminalis does not appear to improve the results for this taxon, possibly due to variability in shell ultrastructure. Nonetheless, the intra-crystalline fraction in gastropods approximates a closed system of amino acids and appears to provide a superior subset of amino acids for geochronological applications. PMID:19684879

  10. Fractionation of Fe isotopes by soil microbes and organic acids

    USGS Publications Warehouse

    Brantley, Susan L.; Liermann, Laura; Bullen, Thomas D.

    2001-01-01

    Small natural variations in Fe isotopes have been attributed to biological cycling. However, without understanding the mechanism of fractionation, it is impossible to interpret such variations. Here we show that the δ56Fe of Fe dissolved from a silicate soil mineral by siderophore-producing bacteria is as much as 0.8% lighter than bulk Fe in the mineral. A smaller isotopic shift is observed for Fe released abiotically by two chelates, and the magnitude of the shift increases with affinity of the ligand for Fe, consistent with a kinetic isotope effect during hydrolysis of Fe at the mineral surface. Fe dissolved abiotically without chelates shows no isotopic shift. The δ56Fe of the exchange fraction on soil grains is also lighter by ~0.6%-1% than Fe from both hornblende and iron oxyhydroxides. The kinetic isotope effect is therefore preserved in open systems such as soils. when recorded in the rock record, Fe isotopic fractionation could document Fe transport by organic molecules or by microbes where such entities were present in the geologic past.

  11. Fractional radiofrequency combined with sonophoresis to facilitate skin penetration of 5-aminolevulinic acid.

    PubMed

    Park, Jong Min; Jeong, Ki-Heon; Bae, Myong Il; Lee, Sang-Jun; Kim, Nack-In; Shin, Min Kyung

    2016-01-01

    Ablative fractional technology has been used to improve transdermal drug delivery. However, there have been few previous in vivo investigations of the relative potency and methodology of fractional radiofrequency (RF) combined with sonophoresis. The purpose of this study was to investigate the effects of fractional RF combined with sonophoresis on 5-aminolevulinic acid (ALA) penetration of the skin. Three male domestic swine were used. The skin of the pigs was exposed to fractional RF and/or sonophoresis, followed by topical ALA application. Fluorescence intensity (FI) of porphyrin fluorescence was measured. In both the epidermis and the dermis, FI increased after fractional RF and increased additionally with the addition of sonophoresis. Fractional RF with sonophoresis effectively enhanced ALA skin penetration. Pre-fractional RF followed by posttreatment with sonophoresis can be used for ALA-photodynamic therapy to achieve higher ALA uptake.

  12. Spectroscopic study of humic acids fractionated by means of tangential ultrafiltration

    NASA Astrophysics Data System (ADS)

    Francioso, O.; Sánchez-Cortés, S.; Casarini, D.; Garcia-Ramos, J. V.; Ciavatta, C.; Gessa, C.

    2002-05-01

    Different chemical and spectroscopic techniques—diffuse reflectance infrared Fourier transform (DRIFT), surface-enhanced Raman spectroscopy (SERS), and 1H, 13C nuclear magnetic resonance (NMR) have been applied to investigate a peat humic acid (HA) separated by tangential ultrafiltration into different nominal molecular weight (NMW) fractions. Each fraction analyzed showed a characteristic DRIFT and NMR pattern. High nominal molecular weight fractions were mainly characterized by long chains of methyl and methylene groups and poorly substituted aromatic rings, while in low nominal molecular weight fractions (L-NMW), phenolic and oxygen-containing groups were predominant. A comparative study on fractions before and after treatment with 0.5 M HCl was carried out. Purified fractions showed either an increase in the carboxylate and phenolic OH groups or an improvement in signal-to-noise ratio of their NMR spectra. The SERS study of NMW fractions allowed significative information on structure and conformation of these fractions. In particular, L-NMW fractions showed a great structural modification, when different alkaline extractants or treatment with HCl were used. Humic-like substances obtained by catechol and gallic acid polymerization on metal surface were investigated using SERS. The SERS spectra of these polymers were compared and discussed with those of NMW HA fractions.

  13. Characterization of humic acid fractions by C-13 nuclear magnetic resonance spectroscopy

    USGS Publications Warehouse

    Wershaw, R. L.; Thorn, K.A.; Pinckney, D.J.

    1988-01-01

    Soil humic acids from different environments were fractionated by adsorption chromatography on Sephadex and characterized by C-13 nuclear magnetic resonance (NMR) spectroscopy. The C-13 NMR spectra of the fractions consist of some sharp, well-resolved lines and some broad bands in contrast to the spectra of the unfractionated humic acids, where the bands are broader and less well-resolved. The marked increase in resolution is apparently due to increased homogeneity of the fractions. These spectra are compared to the spectra of model compounds.

  14. Hydroxylamine hydrochloride-acetic acid-soluble and -insoluble fractions of pelagic sediment: Readsorption revisited

    USGS Publications Warehouse

    Piper, D.Z.; Wandless, G.A.

    1992-01-01

    The extraction of the rare earth elements (REE) from deep-ocean pelagic sediment, using hydroxylamine hydrochloride-acetic acid, leads to the separation of approximately 70% of the bulk REE content into the soluble fraction and 30% into the insoluble fraction. The REE pattern of the soluble fraction, i.e., the content of REE normalized to average shale on an element-by-element basis and plotted against atomic number, resembles the pattern for seawater, whereas the pattern, as well as the absolute concentrations, in the insoluble fraction resembles the North American shale composite. These results preclude significant readsorption of the REE by the insoluble phases during the leaching procedure.

  15. Inhibition of Staphylococcus aureus by crude and fractionated extract from lactic acid bacteria.

    PubMed

    Wong, C-B; Khoo, B-Y; Sasidharan, S; Piyawattanametha, W; Kim, S H; Khemthongcharoen, N; Ang, M-Y; Chuah, L-O; Liong, M-T

    2015-03-01

    Increasing levels of antibiotic resistance by Staphylococcus aureus have posed a need to search for non-antibiotic alternatives. This study aimed to assess the inhibitory effects of crude and fractionated cell-free supernatants (CFS) of locally isolated lactic acid bacteria (LAB) against a clinical strain of S. aureus. A total of 42 LAB strains were isolated and identified from fresh vegetables, fresh fruits and fermented products prior to evaluation of inhibitory activities. CFS of LAB strains exhibiting a stronger inhibitive effect against S. aureus were fractionated into crude protein, polysaccharide and lipid fractions. Crude protein fractions showed greater inhibition against S. aureus compared to polysaccharide and lipid fractions, with a more prevalent effect from Lactobacillus plantarum 8513 and L. plantarum BT8513. Crude protein, polysaccharide and lipid fractions were also characterised with glycine, mannose and oleic acid being detected as the major component of each fraction, respectively. Scanning electron microscopy revealed roughed and wrinkled membrane morphology of S. aureus upon treatment with crude protein fractions of LAB, suggesting an inhibitory effect via the destruction of cellular membrane. This research illustrated the potential application of fractionated extracts from LAB to inhibit S. aureus for use in the food and health industry.

  16. Organic amendment effects on the transformation and fractionation of aluminum in acidic sandy soil

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study was attempted to evaluate the transformation of aluminum (Al) in an acidic sandy soil amended with composts (yard waste, yard + municipal waste, GreenEdge®, and synthetic humic acid), based on soil Al fractionation by single and sequential extraction. The compost amendment significantly i...

  17. CHARACTERIZATION OF HUMIC ACID SIZE FRACTIONS BY SEC AND MALS (R822832)

    EPA Science Inventory

    Latahco silt-loam humic acid was separated on a preparatory scale by size exclusion chromatography (SEC) on a gravity-fed Sepharose column. Four fractions from this separation were collected and further analyzed, along with whole humic acid, by high-performance SEC coupled with a...

  18. A novel solid-state fractionation of naphthenic acid fraction components from oil sands process-affected water.

    PubMed

    Mohamed, Mohamed H; Wilson, Lee D; Shah, Jaimin R; Bailey, Jon; Peru, Kerry M; Headley, John V

    2015-10-01

    Various sorbent materials were evaluated for the fractionation of naphthenic acid fraction components (NAFCs) from oil sand process-affected water (OSPW). The solid phase materials include activated carbon (AC), cellulose, iron oxides (magnetite and goethite), polyaniline (PANI) and three types of biochar derived from biomass (BC-1; rice husks, BC-2; acacia low temperature and BC-3; acacia high temperature). NAFCs were semi-quantified using electrospray ionization high resolution Orbitrap mass spectrometry (ESI-MS) and the metals were assessed by inductively coupled plasma optical emission spectrometry (ICP-OES). The average removal efficacy of NAFCs by AC was 95%. The removal efficacy decreased in the following order: AC, BC-1>BC-2, BC-3, goethite>PANI>cellulose, magnetite. The removal of metals did not follow a clear trend; however, there was notable leaching of potassium by AC and biochar samples. The bound NAFCs by AC were desorbed efficiently with methanol. Methanol regeneration and recycling of AC revealed 88% removal on the fourth cycle; a 4.4% decrease from the first cycle. This fractionation method represents a rapid, cost-effective, efficient, and green strategy for NAFCs from OSPW, as compared with conventional solvent extraction.

  19. Sequential fractionation with concurrent chemical and toxicological characterization of the combustion products of chlorogenic acid.

    PubMed

    Kaur, Navneet; Lacasse, Martine; Fürtös, Alexandra; Waldron, Karen C; Morin, André

    2009-06-05

    Chlorogenic acid is the most abundant polyphenol found in the tobacco plant. The biological effects of its combustion products remain largely unknown. In this study, chlorogenic acid was burned at 640 degrees C for 2 min and the particulate matter of the smoke was collected onto Cambridge filter pads followed by selective extraction in five different solvents. Various fractions of the chlorogenic acid combustion products were tested for induction of micronuclei in V79 Chinese hamster fibroblast cells. Over 40 compounds were identified in the dimethyl sulfoxide (DMSO) extract by high-performance liquid chromatography coupled to electrospray time-of-flight mass spectrometry (HPLC/TOF-MS). The DMSO extract was then fractionated into three major fractions by preparative LC. The fraction inducing the highest degree of toxicity was further separated into four sub-fractions. The sub-fraction responsible for the most toxic response was determined to contain catechol as its major component. The overall reproducibility of the combustion, the extraction procedure and the chemical characterization of the compounds responsible for the toxicity in the chlorogenic acid smoke were evaluated by LC/TOF-MS.

  20. Acid-catalyzed hydrothermal severity on the fractionation of agricultural residues for xylose-rich hydrolyzates.

    PubMed

    Lee, Ji Ye; Ryu, Hyun Jin; Oh, Kyeong Keun

    2013-03-01

    The objective of this work was to investigate the feasibility of acid-catalyzed hydrothermal fractionation for maximum solubilization of the hemicellulosic portion of three agricultural residues. The fractionation conditions converted into combined severity factor (CS) in the range of 1.2-2.9. The highest hemicellulose yield of 87.88% was achieved when barley straw was fractionated at a CS of 2.19. However, the maximum glucose release of 15.29% was achieved for the case of rice straw. The maximum productions of various by-products were observed with the fractionation of rape straw: 0.88 g/L of 5-hydroxymethylfurfural (5-HMF), 2.16 g/L of furfural, 0.44 g/L of levulinic acid, 1.59 g/L of formic acid, and 3.06 g/L of acetic acid. The highest selectivities, a criterion for evaluating the fractionation of 21.55 for fractionated solid and 7.48 for liquid hydrolyzate were obtained from barley straw.

  1. Characterization of pH-fractionated humic acids derived from Chinese weathered coal.

    PubMed

    Zhang, Shuiqin; Yuan, Liang; Li, Wei; Lin, Zhian; Li, Yanting; Hu, Shuwen; Zhao, Bingqiang

    2017-01-01

    To reduce the compositional and structural heterogeneity of humic acids (HAs) and achieve better use of HA resources, in this study, we report a new sequential dissolution method for HAs derived from Chinese weathered coal. This method was used to separate HAs into seven fractions by adjusting the pH (3-10) of the extraction solution. The results showed that the HA fractions derived from Chinese weathered coal were concentrated up to 90.31% in the lower pH solutions (3-7). The compositional and structural characteristics of the HA fractions were determined by elemental analysis; ultraviolet-visible (UV-Vis), Fourier transform infrared (FTIR), and solid-state (13)C-nuclear magnetic resonance (NMR) spectroscopies; and other techniques. The results showed significant differences among the HA fractions. The concentrations of the total acidic groups and the carboxyl groups decreased with the increasing pH of the extraction solution. However, the HA fractions derived from extraction solutions with pH 3-4 had relatively lower aromaticity but a higher protonated carbon content. The HA fractions derived from extraction solutions with pH 6-7 had the highest aromaticity and the greatest abundance of COO/N-C=O. This study demonstrated that adjusting the pH of the extraction solution is one way to fractionate HAs from Chinese weathered coal and to obtain HA fractions with compositions and structures that could serve as useful material for study and utilization.

  2. Temperature dependence of oxygen isotope acid fractionation for modern and fossil tooth enamels.

    PubMed

    Passey, Benjamin H; Cerling, Thure E; Levin, Naomi E

    2007-01-01

    The oxygen isotope ratio of CO(2) liberated from structural carbonate in tooth enamel apatite was measured at phosphoric acid reaction temperatures of 25 degrees C, 60 degrees C and 90 degrees C, and it was found that apparent acid fractionation factors for pristine enamel, fossilized enamel, and calcite follow different temperature relationships. Using sealed vessel reactions normalized to alpha(25) = 1.01025 (the fractionation factor for calcite at 25 degrees C), the apparent fractionation factor at 90 degrees C (alpha*(90)) for pristine enamel ranged between 1.00771 and 1.00820, and between 1.00695 and 1.00772 for fossilized enamel. Apparent fractionation factors for common acid bath reactions are similar to those for sealed vessel reactions. A significant correlation exists between alpha*(90) and F(-) content, suggesting that change in the acid fractionation factor may be related to the replacement of OH(-) with F(-) during fossilization of bioapatite. These results have important implications for making accurate comparisons between modern and fossil tooth enamel delta(18)O values, and for the uniformity of isotope data produced in different laboratories using different acid reaction temperatures.

  3. Highly efficient organosolv fractionation of cornstalk into cellulose and lignin in organic acids.

    PubMed

    Shui, Tao; Feng, Shanghuan; Yuan, Zhongshun; Kuboki, Takashi; Xu, Chunbao Charles

    2016-10-01

    In this study, effects of fractionation solvents, catalysts, temperatures and residence time on yields, purity and chemical composition of the products were investigated at the solid/solvent ratio of 1:5 (g/g). It was revealed that mixture of acetic acid/formic acid/water at the ratio of 3:6:1 (v/v/v) resulted in crude cellulose and lignin products of relatively high purity. The use of HCl catalyst contributed to a high crude cellulose yield, while H2SO4 showed an adverse effect on cellulose yield. However, both of these acidic catalysts contributed to much lower hemicellulose contents in the resulted crude cellulose products compared with those obtained without a catalyst. Fractionation at 90°C for 180min in mixed solvents of acetic acid/formic acid/water (3:6:1, v/v/v) with or without catalyst produced crude cellulose with very low residual lignin contents (<4%).

  4. Combined measurement and significance of lipid-bound sialic acid and carcinoembryonic antigen in detection of human cancer.

    PubMed

    Munjal, D D; Picken, J; Pritchard, J

    1984-01-01

    We evaluated the clinical usefulness of lipid-bound sialic acid (LSA) as a "tumor marker" and assessed individual and carcinoembryonic antigen (CEA) in cancer patients. Serum LSA and CEA concentrations were measured by the resorcinol method after total lipid extraction and isolation of the sialolipid fraction, and by Abbott enzyme immunoassay procedures, respectively. Results indicate that the frequency of elevation and mean LSA values were highest in patients with lung cancer (318 mg/liter), intermediate in miscellaneous (210 mg/liter) and colorectal cancers (200 mg/liter), and lowest in breast cancer (175 mg/liter); while mean CEA values were highest in colorectal cancer (162.5 micrograms/liter), followed by lung (33.8 micrograms/liter), miscellaneous (30.3 micrograms/liter), and breast cancers (11.6 micrograms/liter). Statistically, LSA and CEA values for cancer patients were significantly (P less than 0.001) higher than for normal subjects. The combined measurement of LSA and CEA in serum provides better detection potential for cancer patients than either of the two markers alone.

  5. Proteaselike sequence in hepatitis B virus core antigen is not required for e antigen generation and may not be part of an aspartic acid-type protease.

    PubMed Central

    Nassal, M; Galle, P R; Schaller, H

    1989-01-01

    The hepatitis B virus (HBV) C gene directs the synthesis of two major gene products: HBV core antigen (HBcAg[p21c]), which forms the nucleocapsid, and HBV e antigen (HBeAg [p17e]), a secreted antigen that is produced by several processing events during its maturation. These proteins contain an amino acid sequence similar to the active-site residues of aspartic acid and retroviral proteases. On the basis of this sequence similarity, which is highly conserved among mammalian hepadnaviruses, a model has been put forward according to which processing to HBeAg is due to self-cleavage of p21c involving the proteaselike sequence. Using site-directed mutagenesis in conjunction with transient expression of HBV proteins in the human hepatoma cell line HepG2, we tested this hypothesis. Our results with HBV mutants in which one or two of the conserved amino acids have been replaced by others suggest strongly that processing to HBeAg does not depend on the presence of an intact proteaselike sequence in the core protein. Attempts to detect an influence of this sequence on the processing of HBV P gene products into enzymatically active viral polymerase also gave no conclusive evidence for the existence of an HBV protease. Mutations replacing the putatively essential aspartic acid showed little effect on polymerase activity. Additional substitution of the likewise conserved threonine residue by alanine, in contrast, almost abolished the activity of the polymerase. We conclude that an HBV protease, if it exists, is functionally different from aspartic acid and retroviral proteases. Images PMID:2657101

  6. FT-IR spectra of alginic acid block fractions in three species of brown seaweeds.

    PubMed

    Leal, David; Matsuhiro, Betty; Rossi, Miriam; Caruso, Francesco

    2008-02-04

    Sodium alginates obtained by alkaline extraction of Lessonia flavicans, Desmarestia ligulata and Desmarestia distans (Phaeophyta) from southern Chile were partially hydrolyzed with HCl. Each alginate gave three fractions that were characterized using FT-IR spectroscopy. The fractions soluble in 0.3M HCl presented in the fingerprint region four vibrations at around 960, 911, 890 and 815 cm(-1) that were assigned to heteropolymeric blocks. The fractions soluble at pH 2.85 showed bands at around 948, 888 and 820 cm(-1) attributed to homopolymannuronic acid blocks, the first band is resolved in the second-derivative spectra into two bands at 951 and 936 cm(-1). The fractions insoluble at pH 2.85 presented four bands at around 947, 903, 812 and 781 cm(-1), which were assigned to homopolyguluronic acid blocks. For some samples, the second derivative FT-IR spectra showed new bands indicating the presence of other structures, in low proportions. Structures deduced by FT-IR spectroscopy were corroborated by solution (1)H and (13)C NMR spectroscopy. Two-dimensional spectra were collected to confirm the fine structure of the hetero- and homopolymeric fractions. A geometrically optimized model for the disaccharide alpha-l-gulopyranuronosyl-(1-->4)-alpha-l-gulopyranuronic acid was calculated using density functional theory; good agreement was obtained between its corresponding calculated vibrations and the experimental bands assigned to homopolyguluronic acid blocks.

  7. Improving stable carbon and oxygen isotope geochemical measurements in dolomite: reference material and acid fractionation factor

    NASA Astrophysics Data System (ADS)

    Vandeginste, V.; John, C. M.; Jourdan, A.; Davis, S.

    2010-12-01

    The analysis of stable carbon and oxygen isotope composition is one of the most commonly used techniques in stratigraphic and diagenetic research of carbonate rocks. The wide-spread use and easy access of this long-established method has the side effect that little attention is paid to fundamental calibrations. Dolomite is often measured against a calcite standard (NBS19), and the acid fractionation factor used to calibrate is based on the one for calcite. To date, no reference material exists for dolomite. In this study, which is part of dolomite research in the Qatar Carbonates and Carbon Storage Research Centre project, we focus on two main goals. First, we characterize a current standard of dolomite used for major and minor elemental geochemistry, and assess its suitability as a new dolomite standard for δ18O and δ13C. Second, we attempt to better constrain the acid fractionation factor for dolomite and assess the influence of different dolomite types on this fractionation factor. As only two third of the total oxygen in the carbonate is released in the form of CO2 during acid reaction, a fractionation between the reacting carbonate and the resulting gas will occur. A recent study by Kim et al. (2007) improved on the acid fractionation factors for calcite and aragonite. Often, the acid fractionation factor for dolomite is used to calculate δ18O and δ13C from the values obtained by calibration with the calcite standard. Only two studies (from the 1980’s) have attempted to constrain the acid fractionation factor for dolomite, of which only Rosenbaum and Sheppard (1986) did experiments not only at 25°C, but also at 50 and 100°C. The dataset of the latter authors is, however, very limited and contains only two dolomite samples. We aim at improving the constraints on the acid fractionation factor of dolomite by reacting a wide range of different types of dolomite at a wide range of acid temperature, and compare this to the absolute isotopic composition of

  8. Anti-anaphylactic action of nordihydroguaiaretic acid in antigen sensitized guinea pigs.

    PubMed

    Bergren, Dale R; Valentine, Jimmie L

    2016-12-01

    Therapeutic natural products and medicinal herbs has gained popularity. The anti-antigenic action of the plant alkaloid nordihydroguaiaretic acid (NDGA) was studied in ovalbumin (OA)-sensitized guinea pigs. In one series of experiments conscious, non-sedated guinea pigs were challenged with OA aerosol. Specific airway resistance (SRAW) was monitored using a two-chambered whole-body plethysmograph. OA aerosol increased SRAW above that produced by vehicle administration. Prior NDGA administration by a 1min 0.9% aerosol (w/vol) attenuated the increase in SRAW resulting from OA challenge. In the anesthetized guinea pig pretreated with indomethacin, pyrilamine and propranolol, intravenous OA injection increased intra-tracheal pressure above vehicle injection. Intravenous NDGA administration (5mg/kg) reduced the intra-tracheal pressure increases. In a third series of experiments plasma leukotriene C4 was measured by radio-immunoassay in 3 groups challenged with OA aerosol: vehicle-treated OA-sensitized, OA-sensitized receiving NDGA and vehicle treated guinea pigs. NDGA pretreatment reduced plasma LTC4 in response to OA challenge in OA sensitized guinea pigs. This study demonstrates that NDGA is an effective antigenic agent when given by aerosol or intravenous injection in either conscious or anesthetized guinea pigs, respectively. The mechanism of action of NDGA is presumed primarily be due to the blockage of 5-lipoxygenase and therefore the synthesis of leukotrienes.

  9. Potent and Selective Peptidyl Boronic Acid Inhibitors of the Serine Protease Prostate-Specific Antigen

    PubMed Central

    LeBeau, Aaron M.; Singh, Pratap; Isaacs, John T.; Denmeade, Samuel R.

    2012-01-01

    SUMMARY Prostate cancer cells produce high (microgram to milligram/milliliter) levels of the serine protease Prostate-Specific Antigen (PSA). PSA is enzymatically active in the extracellular fluid surrounding prostate cancers but is found at 1,000- to 10,000-fold lower concentrations in the circulation, where it is inactivated due to binding to abundant serum protease inhibitors. The exclusive presence of high levels of active PSA within prostate cancer sites makes PSA an attractive candidate for targeted imaging and therapeutics. A synthetic approach based on a peptide substrate identified first peptide aldehyde and then boronic acid inhibitors of PSA. The best of these had the sequence Cbz-Ser-Ser-Lys-Leu-(boro)Leu, with a Ki for PSA of 65 nM. The inhibitor had a 60-fold higher Ki for chymotrypsin. A validated model of PSA’s catalytic site confirmed the critical interactions between the inhibitor and residues within the PSA enzyme. PMID:18635003

  10. Augmentation of antigen-specific antibody production and IL-10 generation with a fraction from Rooibos (Aspalathus linearis) tea.

    PubMed

    Ichiyama, Kenji; Tai, Akihiro; Yamamoto, Itaru

    2007-02-01

    Rooibos tea was extracted with boiling water. The aqueous extract was chromatographed in a Diaion HP20 column eluted stepwise with water, 25%, 50% and 75% (v/v) aqueous methanol, and 100% methanol. The water eluate (fraction A) showed an augmenting effect on anti-ovalbumin (anti-OVA) immunoglobulin M (IgM) production in OVA-stimulated murine splenocytes in vitro. Fraction A also showed a strong augmenting effect on interleukin-10 generation in murine splenocytes. Furthermore, continuous ingestion of fraction A was found to increase the anti-OVA IgM level in the sera of OVA-immunized mice.

  11. Kinetics, aggregation behavior and optimization of the fractionation of whey protein isolate with hydrochloric acid

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Concentrated WPI solutions (10% (w/w)) containing approximately 30% alpha-lactalbumin (alpha-LA) and 60% beta-lactoglobulin (beta-LG) were fractionated with HCl at acidic pH and moderate temperatures to denature alpha-LA and recover the alpha-LA aggregates via centrifugation. Aggregation behavior an...

  12. Experimental oxygen isotope fractionation between siderite-water and phosphoric acid liberated CO2-siderite

    USGS Publications Warehouse

    Carothers, W.W.; Adami, L.H.; Rosenbauer, R.J.

    1988-01-01

    The equilibrium fractionation of O isotopes between synthetic siderite and water has been measured at temperatures ranging from 33?? to 197??C. The fractionation between siderite and water over this temperature range can be represented by the equation: 103 ln ?? = 3.13 ?? 106T-2 - 3.50. Comparison between the experimental and theoretical fractionations is favorable only at approximately 200??C; at lower temperatures, they generally differ by up to 2 permil. Siderite was prepared by the slow addition of ferrous chloride solutions to sodium bicarbonate solutions at the experimental temperatures. It was also used to determine the O isotope fractionation factors between phosphoric acid liberated CO2 and siderite. The fractionation factors for this pair at 25?? and 50??C are 1.01175 and 1.01075, respectively. Preliminary results of the measured C isotope fractionation between siderite and Co2 also indicate C isotopic equilibrium during precipitation of siderite. The measured distribution of 13C between siderite and CO2 coincides with the theoretical values only at about 120??C. Experimental and theoretical C fractionations differ up to 3 permil at higher and lower temperatures. ?? 1988.

  13. Taenia saginata metacestode antigenic fractions obtained by ion-exchange chromatography: potential source of immunodominant markers applicable in the immunodiagnosis of human neurocysticercosis.

    PubMed

    Nunes, Daniela da Silva; Gonzaga, Henrique Tomaz; Ribeiro, Vanessa da Silva; da Cunha, Jair Pereira; Costa-Cruz, Julia Maria

    2013-05-01

    The aim of this study was to fractionate and partially characterize fractions obtained from the total saline extract (SE) of Taenia saginata metacestodes after ion-exchange procedure in carboxymethyl sepharose (CM) and diethylaminoethyl sepharose (DEAE) resins, as a source of antigenic markers applicable in the immunodiagnosis of neurocysticercosis (NCC). For IgG detection by enzyme-linked immunosorbent assay (ELISA) and immunoblotting, 140 serum samples were analyzed: 45 from patients with NCC (G1), 50 from patients with other parasitic infections (G2), and 45 from healthy individuals. Sensitivity (Se), specificity (Sp), area under curve (AUC), and likelihood ratios (LR) were calculated. CM S2 and DEAE S2 fractions provided high diagnostic values (Se 88.8% and 93.4%; Sp 93.7% and 92.6%; AUC 0.965 and 0.987; LR+ 14.07 and 12.67; LR- 0.11 and 0.07, respectively). In conclusion, CM S2 and DEAE S2 fractions are important sources of specific peptides, with high efficiency to diagnose NCC.

  14. Catalytic pyrolysis of oil fractions separated from food waste leachate over nanoporous acid catalysts.

    PubMed

    Kim, Seung-Soo; Heo, Hyeon Su; Kim, Sang Guk; Ryoo, Ryong; Kim, Jeongnam; Jeon, Jong-Ki; Park, Sung Hoon; Park, Young-Kwon

    2011-07-01

    Oil fractions, separated from food waste leachate, can be used as an energy source. Especially, high quality oil can be obtained by catalytic cracking. In this study, nanoporous catalysts such as Al-MCM-41 and mesoporous MFI type zeolite were applied to the catalytic cracking of oil fractions using the pyrolysis gas chromatography/mass spectrometry. Mesoporous MFI type zeolite showed better textural porosity than Al-MCM-41. In addition, mesoporous MFI type zeolite had strong Brönsted acidity while Al-MCM-41 had weak acidity. Significant amount of acid components in the food waste oil fractions were converted to mainly oxygenates and aromatics. As a result of its well-defined nanopores and strong acidity, the use of a mesoporous MFI type zeolite produced large amounts of gaseous and aromatic compounds. High yields of hydrocarbons within the gasoline range were also obtained in the case of mesoporous MFI type zeolite, whereas the use of Al-MCM-41, which exhibits relatively weak acidity, resulted in high yields of oxygenates and diesel range hydrocarbons.

  15. Formation of haloacetic acids from dissolved organic matter fractions during chloramination.

    PubMed

    Hong, Ying; Song, Hocheol; Karanfil, Tanju

    2013-03-01

    The objective of this study was to investigate the roles of dissolved organic matter (DOM) fractions, pH and bromide concentration in the formation of haloacetic acids (HAA) during chloramination. DOM from two surface waters with a low (2.9 L/mg-m) and high (5.1 L/mg-m) specific UV absorbance (SUVA(254)) values was isolated and fractionated into three fractions based on the hydrophobicity [i.e., hydrophobic (HPO), transphilic (TPH) and hydrophilic (HPI)]. DOM mass balances and DBP reactivity checks were performed to characterize the effects of isolation and fractionation steps. The fractions were chloraminated at three pHs and three bromide concentrations. The results showed that pH was the most important factor controlling HAA formation and speciation. The HAA yields significantly decreased with increase in pH from 6.3 to 9.0. The impact of bromide in the formation of brominated HAA species also became less important with increasing pH, and no brominated specie was detectable at pH 9. HPO fractions of the two source waters consistently showed higher HAA yields than TPH and HPI fractions. On the other hand, HPI fractions showed higher bromine incorporation than HPO and TPH fractions. To maintain higher and relatively stable combined chlorine residuals while reducing HAA formation, water utilities may consider keeping pH above 7.5 as one strategy. This will also lower the formation of brominated HAA species which have been shown to be more cyto- and geno-toxic than their chlorinated analogs.

  16. Cationic liposome-hyaluronic acid hybrid nanoparticles for intranasal vaccination with subunit antigens

    PubMed Central

    Ochyl, Lukasz J.; Akerberg, Jonathan; Moon, James J.

    2015-01-01

    Here we report the development of a new cationic liposome-hyaluronic acid (HA) hybrid nanoparticle (NP) system and present our characterization of these NPs as an intranasal vaccine platform using a model antigen and F1-V, a candidate recombinant antigen for Yersinia pestis, the causative agent of plague. Incubation of cationic liposomes composed of DOTAP and DOPE with anionic HA biopolymer led to efficient ionic complexation and formation of homogenous liposome-polymer hybrid NPs, as evidenced by fluorescence resonance energy transfer, dynamic light scattering, and nanoparticle tracking analyses. Incorporation of cationic liposomes with thiolated HA allowed for facile surface decoration of NPs with thiol-PEG, resulting in the formation of DOTAP/HA core-PEG shell nanostructures. These NPs, termed DOTAP-HA NPs, exhibited improved colloidal stability and prolonged antigen release. In addition, cytotoxicity associated with DOTAP liposomes (LC50 ~0.2 mg/ml) was significantly reduced by at least 20-fold with DOTAP-HA NPs (LC50 > 4 mg/ml), as measured with bone marrow dendritic cells (BMDCs). Furthermore, NPs co-loaded with ovalbumin (OVA) and a molecular adjuvant, monophosphoryl lipid A (MPLA) promoted BMDC maturation and upregulation of co-stimulatory markers, including CD40, CD86, and MHC-II, and C57BL/6 mice vaccinated with NPs via intranasal route generated robust OVA-specific CD8+ T cell and antibody responses. Importantly, intranasal vaccination with NPs co-loaded with F1-V and MPLA induced potent humoral immune responses with 11-, 23-, and 15-fold increases in F1-V-specific total IgG, IgG1, and IgG2c titers in immune sera by day 77, respectively, and induced balanced Th1/Th2 humoral immune responses, compared with the lack of sero-conversion in mice immunized with the equivalent doses of soluble F1-V vaccine. Overall, these results suggest that liposome-polymer hybrid NPs may serve as a promising vaccine delivery platform for intranasal vaccination against Y

  17. Cationic liposome-hyaluronic acid hybrid nanoparticles for intranasal vaccination with subunit antigens.

    PubMed

    Fan, Yuchen; Sahdev, Preety; Ochyl, Lukasz J; J Akerberg, Jonathan; Moon, James J

    2015-06-28

    Here we report the development of a new cationic liposome-hyaluronic acid (HA) hybrid nanoparticle (NP) system and present our characterization of these NPs as an intranasal vaccine platform using a model antigen and F1-V, a candidate recombinant antigen for Yersinia pestis, the causative agent of plague. Incubation of cationic liposomes composed of DOTAP and DOPE with anionic HA biopolymer led to efficient ionic complexation and formation of homogenous liposome-polymer hybrid NPs, as evidenced by fluorescence resonance energy transfer, dynamic light scattering, and nanoparticle tracking analyses. Incorporation of cationic liposomes with thiolated HA allowed for facile surface decoration of NPs with thiol-PEG, resulting in the formation of DOTAP/HA core-PEG shell nanostructures. These NPs, termed DOTAP-HA NPs, exhibited improved colloidal stability and prolonged antigen release. In addition, cytotoxicity associated with DOTAP liposomes (LC50~0.2mg/ml) was significantly reduced by at least 20-fold with DOTAP-HA NPs (LC50>4mg/ml), as measured with bone marrow derived dendritic cells (BMDCs). Furthermore, NPs co-loaded with ovalbumin (OVA) and a molecular adjuvant, monophosphoryl lipid A (MPLA) promoted BMDC maturation and upregulation of co-stimulatory markers, including CD40, CD86, and MHC-II, and C57BL/6 mice vaccinated with NPs via intranasal route generated robust OVA-specific CD8(+) T cell and antibody responses. Importantly, intranasal vaccination with NPs co-loaded with F1-V and MPLA induced potent humoral immune responses with 11-, 23-, and 15-fold increases in F1-V-specific total IgG, IgG1, and IgG2c titers in immune sera by day 77, respectively, and induced balanced Th1/Th2 humoral immune responses, whereas mice immunized with the equivalent doses of soluble F1-V vaccine failed to achieve sero-conversion. Overall, these results suggest that liposome-polymer hybrid NPs may serve as a promising vaccine delivery platform for intranasal vaccination against Y

  18. Binding characteristics of Cu(2+) to natural humic acid fractions sequentially extracted from the lake sediments.

    PubMed

    He, En; Lü, Changwei; He, Jiang; Zhao, Boyi; Wang, Jinghua; Zhang, Ruiqing; Ding, Tao

    2016-11-01

    Humic acids (HAs) determine the distribution, toxicity, bioavailability, and ultimate fate of heavy metals in the environment. In this work, ten HA fractions (F1-F10) were used as adsorbent, which were sequentially extracted from natural sediments of Lake Wuliangsuhai, to investigate the binding characteristics of Cu(2+) to HA. On the basis of the characterization results, differences were found between the ten extracted HA fractions responding to their elemental compositions and acidic functional groups. The characterization results reveal that the responses of ten extracted HA fractions to their elemental compositions and acidic functional groups were different. The O/C and (O + N)/C ratio of F1-F8 approximately ranged from 0.66 to 0.53 and from 0.72 to 0.61, respectively; the measured results showed that the contents of phenolic groups and carboxyl groups decreased from 4.46 to 2.60 mmol/g and 1.60 to 0.58 mmol/g, respectively. The binding characteristics of Cu(2+) to the ten HA fractions were well modeled by the bi-Langmuir model; the binding behavior of Cu(2+) to all the ten HA fractions were strongly impacted by pH and ionic strength. The FTIR and SEM-EDX image of HA fractions (pre- and post-adsorption) revealed that carboxyl and phenolic groups were responsible for the Cu(2+) sorption on the ten sequentially extracted HA fractions process, which is the same with the analysis of the ligand binding and bi-Langmuir models Accordingly, the adsorption capacity of the former HA fractions on Cu(2+) were higher than the latter ones, which may be attributed to the difference of carboxyl and phenolic group contents between the former and latter extracted HA fractions. Additionally, the functional groups with N and S should not be neglected. This work is hopeful to understand the environmental effect of humic substances, environmental geochemical behavior, and bioavailability of heavy metals in lakes.

  19. Fractionation of Suwannee River fulvic acid and aldrich humic acid on alpha-Al2O3: spectroscopic evidence.

    PubMed

    Claret, Francis; Schäfer, Thorsten; Brevet, Julien; Reiller, Pascal E

    2008-12-01

    Sorptive fractionation of Suwannee River Fulvic Acid (SRFA) and Purified Aldrich Humic Acid (PAHA) on alpha-Al2O3 at pH 6 was probed in the supernatant using different spectroscopic techniques. Comparison of dissolved organic carbon (DOC) analysis with UV/vis spectrophotometric measurements at 254 nm, including specific UV absorbance (SUVA) calculation, revealed a decrease in chromophoric compounds for the nonsorbed extracts after a 24 h contact time. This fractionation, only observable below a certain ratio between initial number of sites of humic substances and of alpha-Al2O3, seems to indicate a higher fractionation for PAHA. C(1s) near-edge X-ray absorption fine structure spectroscopy (NEXAFS) confirmed this trend and points to a decrease in phenolic moieties in the supernatant and to an eventual increase in phenolic moieties on the surface. Time-resolved luminescence spectroscopy (TRLS) of Eu(III) as luminescent probe showed a decrease in the ratio between the (5)D0-->(7)F2 and (5)D0-->(7)F1 transitions for the fractionated organic matter (OM) that is thought to be associated with a lower energy transfer from the OM to Eu(III) due to the loss of polar aromatics. These modifications in the supernatant are a hint for the modification of sorbed humic extracts on the surface.

  20. Fractionation of Suwannee River Fulvic Acid and Aldrich Humic Acid on α-Al2O3: Spectroscopic Evidence

    SciTech Connect

    Claret, F.; Schäfer, T; Brevet, J; Reiller, P

    2008-01-01

    Sorptive fractionation of Suwannee River Fulvic Acid (SRFA) and Purified Aldrich Humic Acid (PAHA) on a-Al2O3 at pH 6 was probed in the supernatant using different spectroscopic techniques. Comparison of dissolved organic carbon (DOC) analysis with UV/vis spectrophotometric measurements at 254 nm, including specific UV absorbance (SUVA) calculation, revealed a decrease in chromophoric compounds for the nonsorbed extracts after a 24 h contact time. This fractionation, only observable below a certain ratio between initial number of sites of humic substances and of a-Al2O3, seems to indicate a higher fractionation for PAHA. C(1s) near-edge X-ray absorption fine structure spectroscopy (NEXAFS) confirmed this trend and points to a decrease in phenolic moieties in the supernatant and to an eventual increase in phenolic moieties on the surface. Time-resolved luminescence spectroscopy (TRLS) of Eu(III) as luminescent probe showed a decrease in the ratio between the 5D0?7F2 and 5D0?7F1 transitions for the fractionated organic matter (OM) that is thought to be associated with a lower energy transfer from the OM to Eu(III) due to the loss of polar aromatics. These modifications in the supernatant are a hint for the modification of sorbed humic extracts on the surface.

  1. Structure and genetics of the O-antigen of Enterobacter cloacae G3054 containing di-N-acetylpseudaminic acid.

    PubMed

    Perepelov, Andrei V; Wang, Min; Filatov, Andrei V; Guo, Xi; Shashkov, Alexander S; Wang, Lei; Knirel, Yuriy A

    2015-04-30

    Mild acid degradation of the lipopolysaccharide of Enterobacter cloacae G3054 resulted in the cleavage of the O-polysaccharide at the linkage of residues of 5,7-diacetamido-3,5,7,9-tetradeoxy-L-glycero-L-manno-non-2-ulosonic acid (di-N-acetylpseudaminic acid, Pse5Ac7Ac) in the main chain. The resultant oligosaccharide and an alkali-treated lipopolysaccharide were studied by sugar analysis along with (1)H and (13)C NMR spectroscopy, and the following structure of the branched pentasaccharide O-unit of the O-polysaccharide was established: [structure: see text] The O-antigen gene cluster of E. cloacae G3054 between conserved genes galF and gnd was sequenced. Most genes necessary for the O-antigen synthesis were found in the cluster and their functions were tentatively assigned by comparison with sequences in the available databases.

  2. A fractional order model for lead-acid battery crankability estimation

    NASA Astrophysics Data System (ADS)

    Sabatier, J.; Cugnet, M.; Laruelle, S.; Grugeon, S.; Sahut, B.; Oustaloup, A.; Tarascon, J. M.

    2010-05-01

    With EV and HEV developments, battery monitoring systems have to meet the new requirements of car industry. This paper deals with one of them, the battery ability to start a vehicle, also called battery crankability. A fractional order model obtained by system identification is used to estimate the crankability of lead-acid batteries. Fractional order modelling permits an accurate simulation of the battery electrical behaviour with a low number of parameters. It is demonstrated that battery available power is correlated to the battery crankability and its resistance. Moreover, the high-frequency gain of the fractional model can be used to evaluate the battery resistance. Then, a battery crankability estimator using the battery resistance is proposed. Finally, this technique is validated with various battery experimental data measured on test rigs and vehicles.

  3. Low acid hydrothermal fractionation of Giant Miscanthus for production of xylose-rich hydrolysate and furfural.

    PubMed

    Kim, Tae Hyun; Ryu, Hyun Jin; Oh, Kyeong Keun

    2016-10-01

    Low acid hydrothermal (LAH) fractionation was developed for the effective recovery of hemicellulosic sugar (mainly xylose) from Miscanthus sacchariflorus Goedae-Uksae 1 (M. GU-1). The xylose yield was maximized at 74.75% when the M. GU-1 was fractionated at 180°C and 0.3wt.% of sulfuric acid for 10min. At this condition, the hemicellulose (mainly xylan) degradation was 86.41%. The difference between xylan degradation and xylose recovery yield, i.e., xylan loss, was 11.66%, as indicated by the formation of decomposed products. The furfural, the value added biochemical product, was also obtained by 0.42g/L at this condition, which was 53.82% of furfural production yield based on the xylan loss. After then, the furfural production continued to increase to a maximum concentration of 1.87g/L, at which point the xylan loss corresponded to 25.87%.

  4. Phosphatidic acid phosphatase activity in subcellular fractions of normal and dystrophic human muscle.

    PubMed

    Kunze, D; Rüstow, B; Olthoff, D; Jung, K

    1985-03-15

    Biopsy samples from normal and dystrophic human muscle (Duchenne type) were fractionated by differential centrifugation and microsomes, mitochondria and cytosol were assayed for phosphatidic acid phosphatase (EC 3.1.3.4) and marker enzymes of mitochondria and cytosol. The activity of phosphatidic acid phosphatase was significantly lower in microsomes and higher in cytosol and mitochondria of dystrophic muscle than in the corresponding subcellular fractions of normal muscle. The results support an explanation of earlier findings that there is reduced G3P incorporation into diglycerides and phosphatidylcholine and a qualitative and quantitative change in the amount of phosphatidylcholine in dystrophic microsomes. The possible reasons for the reduction in the activity of only microsomal PA-P-ase were discussed.

  5. Metabolism of palmitic acid in the subcellular fractions of mouse brain.

    PubMed

    Sun, G Y; Horrocks, L A

    1973-03-01

    After an intracerebral injection of [(14)C]palmitic acid to C57BL/10J mice, the radioactivity in the brains decreased rapidly with time. The incorporated radioactivity was primarily in the 16:0 acyl groups of the diacyl phosphoglycerides at 1 and 3 days after injection. At longer times, increasing proportions of the radioactivity were found in cerebrosides, alkenyl groups, and other acyl groups. The specific radioactivities of the phosphoglycerides were highest in the microsomal fraction at 1 day after injection. The exchange of the diacyl glycerophosphorylcholines and diacyl glycerophosphorylethanolamines between the microsomes and the myelin required 8-14 days. When calculated on the basis of the radioactivity in the 16:0 acyl groups, the half-lives for both of these phosphoglycerides were 6-8 days in all subcellular fractions during the period from 14 to 30 days after injection. The radioactivity in the total lipids from the purified myelin fraction did not decline until more than 14 days after injection because of the reutilization of labeled 16:0 acyl groups for lipid biosynthesis. Recycling of the acyl groups explains the long half-lives reported for myelin phosphoglycerides after injection of [(14)C]acetic acid. Lipids with a relatively high specific radioactivity were lost from the myelin fraction during the purification procedure. The most likely source of these lipids is the most recently formed myelin that is not consolidated into the myelin sheath.

  6. Evidence that the beta-acids fraction of hops reduces central GABAergic neurotransmission.

    PubMed

    Zanoli, P; Zavatti, M; Rivasi, M; Brusiani, F; Losi, G; Puia, G; Avallone, R; Baraldi, M

    2007-01-03

    Humulus lupulus (hops) is traditionally used as a tranquilizing herbal remedy. Here, we investigated the in vivo and in vitro effect of hop beta-acids on central nervous system function. Oral administration of beta-acids (5-10mg/kg) in rats produced an increased exploratory activity in the open field, a reduction in the pentobarbital hypnotic activity and a worsening of picrotoxin-induced seizures. When dosed at 10mg/kg, beta-acids increased, in the elevated plus maze, open arm entries reducing in parallel those in closed arms. In the forced swimming test, we observed a reduction in the immobility time that could suggest an antidepressant-like activity. Electrophysiological studies performed on cerebellar granule cells in culture showed that the beta-acids fraction decreased GABA-evoked current in a dose-dependent way. The effect was not inhibited by the benzodiazepine antagonist Ro 15-1788. Benzodiazepine receptors involvement was also excluded by [(3)H]-Ro 15-1788 binding assay. In conclusion, the behavioral effects of beta-acids fraction could be explained by a reduction in the GABAergic activity although we cannot rule out the involvement of other neurotransmitter systems.

  7. Improved Estimation of Oil, Linoleic and Oleic Acid and Seed Hull Fractions in Safflower by NIRS.

    PubMed

    Rudolphi, Sabine; Becker, Heiko C; Schierholt, Antje; von Witzke-Ehbrecht, Sabine

    2012-03-01

    Near-infrared reflectance spectroscopy (NIRS) of intact seeds allows the non-destructive estimation of seed quality parameters which is highly desirable in plant breeding. Together with yield, oil content and quality, a main aim in safflower (Carthamus tinctorius L.) breeding is the selection of genotypes with a low percentage of empty seeds even under cooler climates. We developed NIRS calibrations for seed oil content, oleic and linoleic acid content, the seed hull fraction and the percentage of empty seeds using seed meal and intact seeds. For the different calibrations 108-534 samples from a safflower breeding program with lines adapted to German conditions, were analyzed with reference analyses (Soxhlet, gas chromatography), and scanned by NIRS as intact seeds and seed meal. Calibration equations were developed and tested through cross validation. The coefficient of determination of the calibration (R(2)) for intact seeds ranged from 0.91(oil content), 0.90 (seed hull fraction), 0.84 (empty seeds), 0.73 (linoleic acid) to 0.68 (oleic acid). The coefficient of determination of the cross validation was higher for seed meal than for intact seeds except for the parameter seed hull fraction. The results show that NIRS calibrations are applicable in safflower breeding programs for a fast screening.

  8. Citric acid mediates the iron absorption from low molecular weight human milk fractions.

    PubMed

    Palika, Ravindranadh; Mashurabad, Purna Chandra; Kilari, Sreenivasulu; Kasula, Sunanda; Nair, Krishnapillai Madhavan; Raghu, Pullakhandam

    2013-11-20

    Previously, we have demonstrated increased iron absorption from low molecular weight (LMW) human milk whey fractions. In the present study, we investigated the effect of heat denaturation, zinc (a competitor of iron), duodenal cytochrome b (DcytB) antibody neutralization and citrate lyase treatment on LMW human milk fraction (>5 kDa referred as 5kF) induced ferric iron reduction, solubilization, and uptake in Caco-2 cells. Heat denaturation and zinc inhibited the 5kF fraction induced ferric iron reduction. In contrast, zinc but not heat denaturation abrogated the ferric iron solubilization activity. Despite inhibition of ferric iron reduction, iron uptake in Caco-2 cells was similar from both native and heat denatured 5kF fractions. However, iron uptake was higher from native compared to heat denatured 5kF fractions in the cells preincubated with the DcytB antibody. Citrate lyase treatment inhibited the ferric iron reduction, solubilization, and uptake in Caco-2 cells. These findings demonstrate that citric acid present in human milk solubilizes the ferric iron which could be reduced by other heat labile components leading to increased uptake in intestinal cells.

  9. A handheld magnetic sensing platform for antigen and nucleic acid detection.

    PubMed

    Pai, Alex; Khachaturian, Aroutin; Chapman, Stephen; Hu, Alexander; Wang, Hua; Hajimiri, Ali

    2014-03-21

    The core requirements for point-of-care (POC) diagnostics necessitate low-cost, portability, easily integrated sample preparation, and quick measurement time. Frequency-shift based magnetic sensing is a measurement technique utilizing a complementary metal-oxide-semiconductor (CMOS) integrated-circuit (IC) chip for magnetic label detection. The sensing scheme leverages the low-cost manufacturing of IC chips while demonstrating the potential for multiplexing capabilities. In this article, we present modifications to this scheme for POC viability. We introduce a handheld reusable reader and a disposable open-well cartridge for the detection of nucleic acids and antigens. The diagnostic system utilizes a novel "magnetic freezing" technique to reduce measurement time, obviates baseline measurement before or during biological assay, and reduces sensor noise. We utilize these enhancements for the room temperature, amplification-free detection of a 31 base-pair DNA oligomer and the interferon-γ (IFN-γ) protein. We have demonstrated reliable measurements down to 100 pM for the DNA assay and 1 pM for the protein.

  10. Lysophosphatidic Acid (LPA) Receptor 5 Inhibits B Cell Antigen Receptor Signaling and Antibody Response1

    PubMed Central

    Shotts, Kristin; Donovan, Erin E.; Strauch, Pamela; Pujanauski, Lindsey M.; Victorino, Francisco; Al-Shami, Amin; Fujiwara, Yuko; Tigyi, Gabor; Oravecz, Tamas; Pelanda, Roberta; Torres, Raul M.

    2014-01-01

    Lysophospholipids have emerged as biologically important chemoattractants capable of directing lymphocyte development, trafficking and localization. Lysophosphatidic acid (LPA) is a major lysophospholipid found systemically and whose levels are elevated in certain pathological settings such as cancer and infections. Here, we demonstrate that BCR signal transduction by mature murine B cells is inhibited upon LPA engagement of the LPA5 (GPR92) receptor via a Gα12/13 – Arhgef1 pathway. The inhibition of BCR signaling by LPA5 manifests by impaired intracellular calcium store release and most likely by interfering with inositol 1,4,5-trisphosphate receptor activity. We further show that LPA5 also limits antigen-specific induction of CD69 and CD86 expression and that LPA5-deficient B cells display enhanced antibody responses. Thus, these data show that LPA5 negatively regulates BCR signaling, B cell activation and immune response. Our findings extend the influence of lysophospholipids on immune function and suggest that alterations in LPA levels likely influence adaptive humoral immunity. PMID:24890721

  11. Human leukocyte antigen haplotype phasing by allele-specific enrichment with peptide nucleic acid probes

    PubMed Central

    Murphy, Nicholas M; Pouton, Colin W; Irving, Helen R

    2014-01-01

    Targeted capture of large fragments of genomic DNA that enrich for human leukocyte antigen (HLA) system haplotypes has utility in haematopoietic stem cell transplantation. Current methods of HLA matching are based on inference or familial studies of inheritance; and each approach has its own inherent limitations. We have designed and tested a probe–target-extraction method for capturing specific HLA haplotypes by hybridization of peptide nucleic acid (PNA) probes to alleles of the HLA-DRB1 gene. Short target fragments contained in plasmids were initially used to optimize the method followed by testing samples of genomic DNA from human subjects with preselected HLA haplotypes and obtained approximately 10% enrichment for the specific haplotype. When performed with high-molecular-weight genomic DNA, 99.0% versus 84.0% alignment match was obtained for the specific haplotype probed. The allele-specific target enrichment that we obtained can facilitate the elucidation of haplotypes between the 65 kb separating the HLA-DRB1 and the HLA-DQA1 genes, potentially spanning a total distance of at least 130 kb. Allele-specific target enrichment with PNA probes is a straightforward technique that has the capability to improve the resolution of DNA and whole genome sequencing technologies by allowing haplotyping of enriched DNA and crucially, retaining the DNA methylation profile. PMID:24936514

  12. Structure-based non-canonical amino acid design to covalently crosslink an antibody-antigen complex.

    PubMed

    Xu, Jianqing; Tack, Drew; Hughes, Randall A; Ellington, Andrew D; Gray, Jeffrey J

    2014-02-01

    Engineering antibodies to utilize non-canonical amino acids (NCAA) should greatly expand the utility of an already important biological reagent. In particular, introducing crosslinking reagents into antibody complementarity determining regions (CDRs) should provide a means to covalently crosslink residues at the antibody-antigen interface. Unfortunately, finding the optimum position for crosslinking two proteins is often a matter of iterative guessing, even when the interface is known in atomic detail. Computer-aided antibody design can potentially greatly restrict the number of variants that must be explored in order to identify successful crosslinking sites. We have therefore used Rosetta to guide the introduction of an oxidizable crosslinking NCAA, l-3,4-dihydroxyphenylalanine (l-DOPA), into the CDRs of the anti-protective antigen scFv antibody M18, and have measured crosslinking to its cognate antigen, domain 4 of the anthrax protective antigen. Computed crosslinking distance, solvent accessibility, and interface energetics were three factors considered that could impact the efficiency of l-DOPA-mediated crosslinking. In the end, 10 variants were synthesized, and crosslinking efficiencies were generally 10% or higher, with the best variant crosslinking to 52% of the available antigen. The results suggest that computational analysis can be used in a pipeline for engineering crosslinking antibodies. The rules learned from l-DOPA crosslinking of antibodies may also be generalizable to the formation of other crosslinked interfaces and complexes.

  13. Amino Acids in Hemagglutinin Antigenic Site B Determine Antigenic and Receptor Binding Differences between A(H3N2)v and Ancestral Seasonal H3N2 Influenza Viruses.

    PubMed

    Wang, Xiaoquan; Ilyushina, Natalia A; Lugovtsev, Vladimir Y; Bovin, Nicolai V; Couzens, Laura K; Gao, Jin; Donnelly, Raymond P; Eichelberger, Maryna C; Wan, Hongquan

    2017-01-15

    Influenza A H3N2 variant [A(H3N2)v] viruses, which have caused human infections in the United States in recent years, originated from human seasonal H3N2 viruses that were introduced into North American swine in the mid-1990s, but they are antigenically distinct from both the ancestral and current circulating H3N2 strains. A reference A(H3N2)v virus, A/Minnesota/11/2010 (MN/10), and a seasonal H3N2 strain, A/Beijing/32/1992 (BJ/92), were chosen to determine the molecular basis for the antigenic difference between A(H3N2)v and the ancestral viruses. Viruses containing wild-type and mutant MN/10 or BJ/92 hemagglutinins (HAs) were constructed and probed for reactivity with ferret antisera against MN/10 and BJ/92 in hemagglutination inhibition assays. Among the amino acids that differ between the MN/10 and BJ/92 HAs, those in antigenic site A had little impact on the antigenic phenotype. Within antigenic site B, mutations at residues 156, 158, 189, and 193 of MN/10 HA to those in BJ/92 switched the MN/10 antigenic phenotype to that of BJ/92. Mutations at residues 156, 157, 158, 189, and 193 of BJ/92 HA to amino acids present in MN/10 were necessary for BJ/92 to become antigenically similar to MN/10. The HA amino acid substitutions responsible for switching the antigenic phenotype also impacted HA binding to sialyl receptors that are usually present in the human respiratory tract. Our study demonstrates that antigenic site B residues play a critical role in determining both the unique antigenic phenotype and receptor specificity of A(H3N2)v viruses, a finding that may facilitate future surveillance and risk assessment of novel influenza viruses.

  14. Cu(II) binding by a pH-fractionated fulvic acid

    USGS Publications Warehouse

    Brown, G.K.; Cabaniss, S.E.; MacCarthy, P.; Leenheer, J.A.

    1999-01-01

    The relationship between acidity, Cu(II) binding and sorption to XAD resin was examined using Suwannee River fulvic acid (SRFA). The work was based on the hypothesis that fractions of SRFA eluted from an XAD column at various pH's from 1.0 to 12.0 would show systematic variations in acidity and possibly aromaticity which in turn would lead to different Cu(II) binding properties. We measured equilibrium Cu(II) binding to these fractions using Cu2+ ion-selective electrode (ISE) potentiometry at pH 6.0. Several model ligands were also examined, including cyclopentane-1,2,3,4-tetracarboxylic acid (CP-TCA) and tetrahydrofuran-2,3,4,5-tetracarboxylic acid (THF-TCA), the latter binding Cu(II) much more strongly as a consequence of the ether linkage. The SRFA Cu(II) binding properties agreed with previous work at high ionic strength, and binding was enhanced substantially at lower ionic strength, in agreement with Poisson-Boltzmann predictions for small spheres. Determining Cu binding constants (K(i)) by non-linear regression with total ligand concentrations (L(Ti)) taken from previous work, the fractions eluted at varying pH had K(i) similar to the unfractionated SRFA, with a maximum enhancement of 0.50 log units. We conclude that variable-pH elution from XAD does not isolate significantly strong (or weak) Cu(II)-binding components from the SRFA mixture. Copyright (C) 1999 Elsevier Science B.V.

  15. Acid-base and copper-binding properties of three organic matter fractions isolated from a forest floor soil solution

    NASA Astrophysics Data System (ADS)

    van Schaik, Joris W. J.; Kleja, Dan B.; Gustafsson, Jon Petter

    2010-02-01

    Vast amounts of knowledge about the proton- and metal-binding properties of dissolved organic matter (DOM) in natural waters have been obtained in studies on isolated humic and fulvic (hydrophobic) acids. Although macromolecular hydrophilic acids normally make up about one-third of DOM, their proton- and metal-binding properties are poorly known. Here, we investigated the acid-base and Cu-binding properties of the hydrophobic (fulvic) acid fraction and two hydrophilic fractions isolated from a soil solution. Proton titrations revealed a higher total charge for the hydrophilic acid fractions than for the hydrophobic acid fraction. The most hydrophilic fraction appeared to be dominated by weak acid sites, as evidenced by increased slope of the curve of surface charge versus pH at pH values above 6. The titration curves were poorly predicted by both Stockholm Humic Model (SHM) and NICA-Donnan model calculations using generic parameter values, but could be modelled accurately after optimisation of the proton-binding parameters (pH ⩽ 9). Cu-binding isotherms for the three fractions were determined at pH values of 4, 6 and 9. With the optimised proton-binding parameters, the SHM model predictions for Cu binding improved, whereas the NICA-Donnan predictions deteriorated. After optimisation of Cu-binding parameters, both models described the experimental data satisfactorily. Iron(III) and aluminium competed strongly with Cu for binding sites at both pH 4 and pH 6. The SHM model predicted this competition reasonably well, but the NICA-Donnan model underestimated the effects significantly at pH 6. Overall, the Cu-binding behaviour of the two hydrophilic acid fractions was very similar to that of the hydrophobic acid fraction, despite the differences observed in proton-binding characteristics. These results show that for modelling purposes, it is essential to include the hydrophilic acid fraction in the pool of 'active' humic substances.

  16. Interleukin-6 and Cyclooxygenase-2 downregulation by fatty-acid fractions of Ranunculus constantinopolitanus

    PubMed Central

    2009-01-01

    Background Medicinal plants represent alternative means for the treatment of several chronic diseases, including inflammation. The genus Ranunculus, a representative of the Ranunculaceae family, has been reported to possess anti-inflammatory, analgesic, antiviral, antibacterial, antiparasitic and antifungal activities, possibly due to the presence of anemonin and other. Different studies have shown the occurrence of unusual fatty acids (FAs) in Ranunculaceae; however, their therapeutic role has not been investigated. The purpose of this study is to characterize potential anti-inflammatory bioactivities in Ranunculus constantinopolitanus D'Urv., traditionally used in Eastern Mediterranean folk medicine. Methods The aerial part of R. constantinopolitanus was subjected to methanol (MeOH) extraction and solvent fractionation. The bioactive fraction (I.2) was further fractionated using column chromatography, and the biologically active subfraction (Y2+3) was identified using infrared (IR) spectroscopy, nuclear magnetic resonance (NMR) and gas chromatography-mass spectrometry (GC-MS). The effects of I.2 and Y2+3 on cell viability were studied in mouse mammary epithelial SCp2 cells using trypan blue exclusion method. To study the anti-inflammatory activities of I.2 and Y2+3, their ability to reduce interleukin (IL)-6 levels was assessed in endotoxin (ET)-stimulated SCp2 cells using enzyme-linked immunosorbent assay (ELISA). In addition, the ability of Y2+3 to reduce cyclooxygenase (COX)-2 expression was studied in IL-1-treated mouse intestinal epithelial Mode-K cells via western blotting. Data were analyzed by one-way analysis of variance (ANOVA), Student-Newman-Keuls (SNK), Tukey HSD, two-sample t-test and Dunnett t-tests for multiple comparisons. Results The chloroform fraction (I.2) derived from crude MeOH extract of the plant, in addition to Y2+3, a FA mix isolated from this fraction and containing palmitic acid, C18:2 and C18:1 isomers and stearic acid (1:5:8:1 ratio

  17. Optimization of acid hydrolysis from the hemicellulosic fraction of Eucalyptus grandis residue using response surface methodology.

    PubMed

    Canettieri, Eliana Vieira; de Moraes Rocha, George Jackson; de Carvalho, João Andrade; de Almeida e Silva, João Batista

    2007-01-01

    Biotechnological conversion of biomass into fuels and chemicals requires hydrolysis of the polysaccharide fraction into monomeric sugars. Hydrolysis can be performed enzymatically and with dilute or concentrate mineral acids. The present study used dilute sulfuric acid as a catalyst for hydrolysis of Eucalyptus grandis residue. The purpose of this paper was to optimize the hydrolysis process in a 1.4 l pilot-scale reactor and investigate the effects of the acid concentration, temperature and residue/acid solution ratio on the hemicellulose removal and consequently on the production of sugars (xylose, glucose and arabinose) as well as on the formation of by-products (furfural, 5-hydroxymethylfurfural and acetic acid). This study was based on a model composition corresponding to a 2(3) orthogonal factorial design and employed the response surface methodology (RSM) to optimize the hydrolysis conditions, aiming to attain maximum xylose extraction from hemicellulose of residue. The considered optimum conditions were: H(2)SO(4) concentration of 0.65%, temperature of 157 degrees C and residue/acid solution ratio of 1/8.6 with a reaction time of 20 min. Under these conditions, 79.6% of the total xylose was removed and the hydrolysate contained 1.65 g/l glucose, 13.65 g/l xylose, 1.55 g/l arabinose, 3.10 g/l acetic acid, 1.23 g/l furfural and 0.20 g/l 5-hydroxymethylfurfural.

  18. Efficient Fractionation and Analysis of Fatty Acids and their Salts in Fat, Oil and Grease (FOG) Deposits.

    PubMed

    Benecke, Herman P; Allen, Sara K; Garbark, Daniel B

    2017-02-01

    A fractionation methodology of fat, oil and grease (FOG) deposits was developed based on the insolubility of fatty acid salts in dichloromethane (DCM) and the relatively high solubility of fatty acids and triglycerides in DCM. Using this method, coupled with spectral analysis, it was shown that fatty acids rather than fatty acid salts were the predominant species in FOG deposits obtained from three metropolitan locations in the United States and that fatty acid triglycerides were either not detected or were present in very small concentrations. This solubility-based fractionation approach also revealed the presence of nitrogen-containing compounds that had not been previously detected in FOG deposits including peptides and (or) proteins. The comparison of the ratios of stearic acid salts to stearic acid versus the ratio of palmitic acid salts to palmitic acid in FOG deposits may indicate that the initial step in FOG deposit formation is the preferential precipitation of stearic acid salts.

  19. Fractionation of fulvic acid by iron and aluminum oxides: influence on copper toxicity to Ceriodaphnia dubia

    USGS Publications Warehouse

    Smith, Kathleen S.; James F. Ranville,; Emily K. Lesher,; Daniel J. Diedrich,; Diane M. McKnight,; Ruth M. Sofield,

    2014-01-01

    This study examines the effect on aquatic copper toxicity of the chemical fractionation of fulvic acid (FA) that results from its association with iron and aluminum oxyhydroxide precipitates. Fractionated and unfractionated FAs obtained from streamwater and suspended sediment were utilized in acute Cu toxicity tests on ,i>Ceriodaphnia dubia. Toxicity test results with equal FA concentrations (6 mg FA/L) show that the fractionated dissolved FA was 3 times less effective at reducing Cu toxicity (EC50 13 ± 0.6 μg Cu/L) than were the unfractionated dissolved FAs (EC50 39 ± 0.4 and 41 ± 1.2 μg Cu/L). The fractionation is a consequence of preferential sorption of molecules having strong metal-binding (more aromatic) moieties to precipitating Fe- and Al-rich oxyhydroxides, causing the remaining dissolved FA to be depleted in these functional groups. As a result, there is more bioavailable dissolved Cu in the water and hence greater potential for Cu toxicity to aquatic organisms. In predicting Cu toxicity, biotic ligand models (BLMs) take into account dissolved organic carbon (DOC) concentration; however, unless DOC characteristics are accounted for, model predictions can underestimate acute Cu toxicity for water containing fractionated dissolved FA. This may have implications for water-quality criteria in systems containing Fe- and Al-rich sediment, and in mined and mineralized areas in particular. Optical measurements, such as specific ultraviolet absorbance at 254 nm (SUVA254), show promise for use as spectral indicators of DOC chemical fractionation and inferred increased Cu toxicity.

  20. The free fractions of circulating docosahexaenoic acid and eicosapentenoic acid as optimal end-point of measure in bioavailability studies on n-3 fatty acids.

    PubMed

    Scarsi, Claudia; Levesque, Ann; Lisi, Lucia; Navarra, Pierluigi

    2015-05-01

    The high complexity of n-3 fatty acids absorption process, along with the huge amount of endogenous fraction, makes bioavailability studies with these agents very challenging and deserving special consideration. In this paper we report the results of a bioequivalence study between a new formulation of EPA+DHA ethyl esters developed by IBSA Institut Biochimique and reference medicinal product present on the Italian market. Bioequivalence was demonstrated according to the criteria established by the EMA Guideline on the Investigation of Bioequivalence. We found that the free fractions represent a better and more sensitive end-point for bioequivalence investigations on n-3 fatty acids, since: (i) the overall and intra-subject variability of PK parameters was markedly lower compared to the same variability calculated on the total DHA and EPA fractions; (ii) the absorption process was completed within 4h, and the whole PK profile could be drawn within 12-15 h from drug administration.

  1. Stable Strontium Isotopic Fractionation During Sorption onto Magnetic Nano-Humid Acid Coated Iron Oxide Particles

    NASA Astrophysics Data System (ADS)

    Liu, H.-C.; You, C.-F.; Tu, Y.-J.

    2012-04-01

    The mobility of strontium (Sr) through hydrological systems is critically governed by sorption reactions of solid phases such as iron oxides and clay minerals. Inorganic precipitated and bacteriogenic iron oxides are widespread in marine and fresh systems fed by iron-rich supplies, which may impact the global Sr cycle. A series of laboratory batch experiments of Sr sorption onto humid acid coated magnetic nano iron oxides using high purity in-house standard were performed in this study and aimed for gaining a better understanding of mechanisms controlling Sr mobility. The results indicate that Sr sorption is a function of pH, 100 % Sr remains unbounded at pH <4 and more than 85 % absorbed at pH >8. Temperature controlled experiments in the range of 5 - 35 degrees C indicate a positive relationship between sorption capacity and temperature. Measurements of stable Sr isotopes (δ88Sr and δ87Sr) in batch experiments show that lighter isotopes reacted preferentially with iron oxides, and a negligible pH effect on isotopic fractionation. To calculate the degree of the kinetic isotope effect, Rayleigh fractionation model is applied to evaluate the isotope fractionation factor, α is 0.99985 at pH 8.17 (equals to Δ88Sr ~ 0.15 ‰). Temperature exerts only minor isotopic fractionation effect and reveals more importance at lower temperatures. This new finding provides crucial information for a detail evaluation of sorption processes in natural environments.

  2. Fractionation of SWNT/nucleic acid complexes by agarose gel electrophoresis.

    PubMed

    Vetcher, Alexandre A; Srinivasan, Srimeenakshi; Vetcher, Ivan A; Abramov, Semen M; Kozlov, Mikhail; Baughman, Ray H; Levene, Stephen D

    2006-08-28

    We show that aqueous dispersions of single-walled carbon nanotubes (SWNTs), prepared with the aid of nucleic acids (NAs) such as RNA or DNA, can be separated into fractions using agarose gel electrophoresis. In a DC electric field, SWNT/NA complexes migrate in the gel in the direction of positive potential to form well-defined bands. Raman spectroscopy as a function of band position shows that nanotubes having different spectroscopic properties possess different electrophoretic mobilities. The migration patterns for SWNT/RNA and SWNT/DNA complexes differ. Parallel elution of the SWNT/NA complexes from the gel during electrophoresis and subsequent characterization by AFM reveals differences in nanotube diameter, length and curvature. The results suggest that fractionation of nanotubes can be achieved by this procedure. We discuss factors affecting the mobility of the nanotube complexes and propose analytical applications of this technique.

  3. Structure and gene cluster of the O-antigen of Escherichia coli O156 containing a pyruvic acid acetal.

    PubMed

    Duan, Zhifeng; Senchenkova, Sof'ya N; Guo, Xi; Perepelov, Andrei V; Shashkov, Alexander S; Liu, Bin; Knirel, Yuriy A

    2016-07-22

    The lipopolysaccharide of Escherichia coli O156 was degraded under mild acidic and alkaline conditions and the resulting polysaccharides were studied by sugar analysis and (1)H and (13)C NMR spectroscopy. The following structure of the pentasaccharide repeating unit of the O-polysaccharide was established: where Rpyr indicates R-configurated pyruvic acid acetal. Minor O-acetyl groups also were present and tentatively localized on the Gal residues. The gene cluster for biosynthesis of the O-antigen of E. coli O156 was analyzed and shown to be consistent with the O-polysaccharide structure.

  4. Fractionation for further conversion: from raw corn stover to lactic acid

    NASA Astrophysics Data System (ADS)

    He, Ting; Jiang, Zhicheng; Wu, Ping; Yi, Jian; Li, Jianmei; Hu, Changwei

    2016-12-01

    Fractionation is considered to be one promising strategy to utilize raw biomass to its fullest and produce chemicals with high selectivity. Herein, ethanol/H2O (1/1, v/v) co-solvent with 0.050 M oxalic acid is used to simultaneously fractionate 88.0 wt% of hemicellulose and 89.2 wt% of lignin in corn stover, while cellulose is not obviously degraded. H2O dissolves hemicellulose, G unit and those with β-O-4 linkage of lignin; whereas ethanol extracts G and S units as well as the skeleton with β-5 and β-β linkages of lignin. Oxalic acid effectively catalyzes the hydrolysis of hemicellulose and breaks the intermolecular linkages between hemicellulose and lignin, therefore further promotes the release of lignin. The dissolved hemicelluloses derivatives are reprocessed to produce lactic acid obtaining a high yield of 79.6 wt% with 90% selectivity by the catalysis of MgO. The remained cellulose and recovered lignin can be used further as feedstock to produce chemicals.

  5. Fractionation for further conversion: from raw corn stover to lactic acid

    PubMed Central

    He, Ting; Jiang, Zhicheng; Wu, Ping; Yi, Jian; Li, Jianmei; Hu, Changwei

    2016-01-01

    Fractionation is considered to be one promising strategy to utilize raw biomass to its fullest and produce chemicals with high selectivity. Herein, ethanol/H2O (1/1, v/v) co-solvent with 0.050 M oxalic acid is used to simultaneously fractionate 88.0 wt% of hemicellulose and 89.2 wt% of lignin in corn stover, while cellulose is not obviously degraded. H2O dissolves hemicellulose, G unit and those with β-O-4 linkage of lignin; whereas ethanol extracts G and S units as well as the skeleton with β-5 and β-β linkages of lignin. Oxalic acid effectively catalyzes the hydrolysis of hemicellulose and breaks the intermolecular linkages between hemicellulose and lignin, therefore further promotes the release of lignin. The dissolved hemicelluloses derivatives are reprocessed to produce lactic acid obtaining a high yield of 79.6 wt% with 90% selectivity by the catalysis of MgO. The remained cellulose and recovered lignin can be used further as feedstock to produce chemicals. PMID:27917955

  6. Contact allergy to acid and neutral fractions of rosins. Sensitization experiments in guinea pigs and patch testing in patients.

    PubMed

    Karlberg, A T; Boman, A; Holmbom, B; Lidén, C

    1986-01-01

    The allergenicity of two different types of rosins (gum rosin and tall oil rosin) was compared. The rosins were divided into their neutral and acid fractions. The neutral fraction of tall oil rosin failed to induce contact sensitivity in animals tested according to the Guinea pig maximization test method (GPMT). The neutral fraction of gum rosin as well as the two acid fractions gave significant responses. Relatively fewer dermatitis patients reacted to the neutral fraction compared with reactions to the unfractionated gum rosin when patch tested. Fewer reactions to tall oil rosin than to gum rosin (p less than 0.05) were observed. It is concluded that tall oil rosin is less allergenic than gum rosin, which may be due to the absence of allergens in its neutral fraction.

  7. Estimate of colostral immunoglobulin G concentration using refractometry without or with caprylic acid fractionation.

    PubMed

    Morrill, K M; Conrad, E; Polo, J; Lago, A; Campbell, J; Quigley, J; Tyler, H

    2012-07-01

    Our objectives were to evaluate the use of refractometry as a means of estimating immunoglobulin G (IgG) concentration of bovine maternal colostrum (MC) and determine if fractionation of MC using caprylic acid (CA) improved estimates of IgG. Samples (n=85) of MC were collected from a single dairy in California and used to determine the method of CA fraction that produced the best prediction of IgG based on CA fractionation followed by refractometry. Subsequently, samples of MC (n=827) were collected from 67 farms in 12 states to compare refractometry with or without CA fractionation as methods to estimate IgG concentration. Samples were collected from the feeding pool and consisted of fresh (n=196), previously frozen (n=479), or refrigerated (n=152) MC. Samples were further classified by the number freeze-thaw cycles before analysis. Fractionation with CA was conducted by adding 1 mL of MC to a tube containing 75 μL of CA and 1 mL of 0.06 M acetic acid. The tube was shaken and allowed to react for 1 min. Refractive index of the IgG-rich supernatant (nDf) was determined using a digital refractometer. Whole, nonfractionated MC was analyzed for IgG by radial immunodiffusion (RID) and refractive index (nDw). The relationship between nDf and IgG (r=0.53; n=805) was weak, whereas that between nDw and IgG was stronger (r=0.73; n=823). Fresh samples analyzed by refractometry that subsequently went through 1 freeze-thaw cycle before RID analysis resulted in the strongest relationship between IgG and nDf or nDw (r=0.93 and 0.90, respectively). The MC samples collected fresh on the farm but frozen 2 or more times before analysis by refractometry or RID had low correlations between IgG and nDf and nDw (r=0.09 and 0.01). Samples refrigerated or frozen on the farm before analysis had weaker relationships between RID and nDf or nDw (r=0.38 to 0.80), regardless of the number of freeze-thaw cycles. Breed and lactation number did not affect the accuracy of either test. These

  8. Deuterium Fractionation during Amino Acid Formation by Photolysis of Interstellar Ice Analogs Containing Deuterated Methanol

    NASA Astrophysics Data System (ADS)

    Oba, Yasuhiro; Takano, Yoshinori; Watanabe, Naoki; Kouchi, Akira

    2016-08-01

    Deuterium (D) atoms in interstellar deuterated methanol might be distributed into complex organic molecules through molecular evolution by photochemical reactions in interstellar grains. In this study, we use a state-of-the-art high-resolution mass spectrometer coupled with a high-performance liquid chromatography system to quantitatively analyze amino acids and their deuterated isotopologues formed by the photolysis of interstellar ice analogs containing singly deuterated methanol CH2DOH at 10 K. Five amino acids (glycine, α-alanine, β-alanine, sarcosine, and serine) and their deuterated isotopologues whose D atoms are bound to carbon atoms are detected in organic residues formed by photolysis followed by warming up to room temperature. The abundances of singly deuterated amino acids are in the range of 0.3-1.1 relative to each nondeuterated counterpart, and the relative abundances of doubly and triply deuterated species decrease with an increasing number of D atoms in a molecule. The abundances of amino acids increase by a factor of more than five upon the hydrolysis of the organic residues, leading to decreases in the relative abundances of deuterated species for α-alanine and β-alanine. On the other hand, the relative abundances of the deuterated isotopologues of the other three amino acids did not decrease upon hydrolysis, indicating different formation mechanisms of these two groups upon hydrolysis. The present study facilitates both qualitative and quantitative evaluations of D fractionation during molecular evolution in the interstellar medium.

  9. Study of the free volume fraction in polylactic acid (PLA) by thermal analysis

    NASA Astrophysics Data System (ADS)

    Abdallah, A.; Benrekaa, N.

    2015-10-01

    The poly (lactic acid) or polylactide (PLA) is a biodegradable polymer with high modulus, strength and thermoplastic properties. In this work, the evolution of various properties of PLA is studied, such as glass transition temperature, mechanical modules and elongation percentage with the aim of investigating the free volume fraction. To do so, two thermal techniques have been used: the dynamic mechanical analysis (DMA) and dilatometry. The results obtained by these techniques are combined to go back to the structural properties of the studied material.

  10. Acetic acid-assisted hydrothermal fractionation of empty fruit bunches for high hemicellulosic sugar recovery with low byproducts.

    PubMed

    Kim, Dong Young; Um, Byung Hwan; Oh, Kyeong Keun

    2015-07-01

    Xylose, mannose, and galactose (xmg) recovery from empty fruit bunches using acetic acid-assisted hydrothermal (AAH) fractionation method was investigated. Acetic acid has been demonstrated to be effective in xmg recovery in comparison with the liquid hot-water (LHW) fractionation. The maximum xmg recovery yield (50.7 %) from the empty fruit bunch (EFB) was obtained using AAH fractionation at optimum conditions (6.9 wt.% acetic acid at 170 °C and for 18 min); whereas, only 16.2 % of xmg recovery was obtained from the LHW fractionation at the same reaction conditions (170 °C and 18 min). Releasing out the glucose from EFB was kept at low level (<1.0 %) through all tested conditions and consequently negligible 5-HMF and formic acid were analyzed in the hydrolyzate. The production of furfural was also resulted with extremely low level (1.0 g/L).

  11. Identification of Low- and High-Impact Hemagglutinin Amino Acid Substitutions That Drive Antigenic Drift of Influenza A(H1N1) Viruses

    PubMed Central

    Harvey, William T.; Benton, Donald J.; Gregory, Victoria; Hall, James P. J.; Daniels, Rodney S.; Bedford, Trevor; Haydon, Daniel T.; Hay, Alan J.; McCauley, John W.; Reeve, Richard

    2016-01-01

    Determining phenotype from genetic data is a fundamental challenge. Identification of emerging antigenic variants among circulating influenza viruses is critical to the vaccine virus selection process, with vaccine effectiveness maximized when constituents are antigenically similar to circulating viruses. Hemagglutination inhibition (HI) assay data are commonly used to assess influenza antigenicity. Here, sequence and 3-D structural information of hemagglutinin (HA) glycoproteins were analyzed together with corresponding HI assay data for former seasonal influenza A(H1N1) virus isolates (1997–2009) and reference viruses. The models developed identify and quantify the impact of eighteen amino acid substitutions on the antigenicity of HA, two of which were responsible for major transitions in antigenic phenotype. We used reverse genetics to demonstrate the causal effect on antigenicity for a subset of these substitutions. Information on the impact of substitutions allowed us to predict antigenic phenotypes of emerging viruses directly from HA gene sequence data and accuracy was doubled by including all substitutions causing antigenic changes over a model incorporating only the substitutions with the largest impact. The ability to quantify the phenotypic impact of specific amino acid substitutions should help refine emerging techniques that predict the evolution of virus populations from one year to the next, leading to stronger theoretical foundations for selection of candidate vaccine viruses. These techniques have great potential to be extended to other antigenically variable pathogens. PMID:27057693

  12. Reversed-phase high-performance liquid chromatography of the stable electrophoretic fractions of soil humic acids

    NASA Astrophysics Data System (ADS)

    Trubetskoi, O. A.; Trubetskaya, O. E.

    2015-02-01

    Reversed-phase high-performance liquid chromatography (RP-HPLC) has been used for the hydrophobicity analysis of soil humic acids and their stable electrophoretic fractions A, B, and C + D preliminarily prepared by the combination of gel permeation chromatography on Sephadex with polyacrylamide gel electrophoresis. In two humic acid preparations of different genesis, the electrophoretic fraction A of the larger molecular size was the most hydrophobic (60-73% of the fraction was irreversibly adsorbed on a hydrophobic reversed-phase (RF) column C18), and the fraction C + D of the smallest molecular size was the most hydrophilic. The fraction B of medium size occupied an intermediate position (33-47% of the fraction was irreversibly adsorbed on the column). The use of RP-HPLC allowed for the first time detecting the hydrophobic electrophoretic fraction A of the largest molecular size mainly composed of aliphatic long-chained hydrocarbon, protein, and carbohydrate fragments in soil humic acids. Data on the degree of hydrophobicity and the earlier obtained physicochemical characteristics of stable electrophoretic fractions are discussed in terms of the supramolecular and macromolecular structure of soil humic acids.

  13. Sialic acid removal from dendritic cells improves antigen cross-presentation and boosts anti-tumor immune responses

    PubMed Central

    Silva, Mariana; Silva, Zélia; Marques, Graça; Ferro, Tiago; Gonçalves, Márcia; Monteiro, Mauro; van Vliet, Sandra J.; Mohr, Elodie; Lino, Andreia C.; Fernandes, Alexandra R.; Lima, Flávia A.; van Kooyk, Yvette; Matos, Teresa; Tadokoro, Carlos E.; Videira, Paula A.

    2016-01-01

    Dendritic cells (DCs) hold promise for anti-cancer immunotherapy. However, clinically, their efficiency is limited and novel strategies to improve DC-mediated anti-tumor responses are needed. Human DCs display high content of sialic acids, which inhibits their maturation and co-stimulation capacity. Here, we aimed to understand whether exogenous desialylation of DCs improves their anti-tumor immunity. Compared to fully sialylated DCs, desialylated human DCs loaded with tumor-antigens showed enhanced ability to induce autologous T cells to proliferate, to secrete Th1 cytokines, and to specifically induce tumor cell apoptosis. Desialylated DCs showed an increased expression of MHC-I and -II, co-stimulatory molecules and an augmented secretion of IL-12. Desialylated HLA-A*02:01 DCs pulsed with gp100 peptides displayed enhanced peptide presentation through MHC-I, resulting in higher activation ofgp100280–288 specific CD8+ cytotoxic T cells. Desialylated murine DCs also exhibited increased MHC and co-stimulatory molecules and higher antigen cross-presentation via MHC-I. These DCs showed higher ability to activate antigen-specific CD4+ and CD8+ T cells, and to specifically induce tumor cell apoptosis. Collectively, our data demonstrates that desialylation improves DCs' ability to elicit T cell-mediated anti-tumor activity, due to increased MHC-I expression and higher antigen presentation via MHC-I. Sialidase treatment of DCs may represent a technology to improve the efficacy of antigen loaded-DC-based vaccines for anti-cancer immunotherapy. PMID:27203391

  14. Sialic acid removal from dendritic cells improves antigen cross-presentation and boosts anti-tumor immune responses.

    PubMed

    Silva, Mariana; Silva, Zélia; Marques, Graça; Ferro, Tiago; Gonçalves, Márcia; Monteiro, Mauro; van Vliet, Sandra J; Mohr, Elodie; Lino, Andreia C; Fernandes, Alexandra R; Lima, Flávia A; van Kooyk, Yvette; Matos, Teresa; Tadokoro, Carlos E; Videira, Paula A

    2016-07-05

    Dendritic cells (DCs) hold promise for anti-cancer immunotherapy. However, clinically, their efficiency is limited and novel strategies to improve DC-mediated anti-tumor responses are needed. Human DCs display high content of sialic acids, which inhibits their maturation and co-stimulation capacity. Here, we aimed to understand whether exogenous desialylation of DCs improves their anti-tumor immunity. Compared to fully sialylated DCs, desialylated human DCs loaded with tumor-antigens showed enhanced ability to induce autologous T cells to proliferate, to secrete Th1 cytokines, and to specifically induce tumor cell apoptosis. Desialylated DCs showed an increased expression of MHC-I and -II, co-stimulatory molecules and an augmented secretion of IL-12. Desialylated HLA-A*02:01 DCs pulsed with gp100 peptides displayed enhanced peptide presentation through MHC-I, resulting in higher activation ofgp100280-288 specific CD8+ cytotoxic T cells. Desialylated murine DCs also exhibited increased MHC and co-stimulatory molecules and higher antigen cross-presentation via MHC-I. These DCs showed higher ability to activate antigen-specific CD4+ and CD8+ T cells, and to specifically induce tumor cell apoptosis. Collectively, our data demonstrates that desialylation improves DCs' ability to elicit T cell-mediated anti-tumor activity, due to increased MHC-I expression and higher antigen presentation via MHC-I. Sialidase treatment of DCs may represent a technology to improve the efficacy of antigen loaded-DC-based vaccines for anti-cancer immunotherapy.

  15. Fractionation of Eucalyptus grandis chips by dilute acid-catalysed steam explosion.

    PubMed

    Emmel, Alexandre; Mathias, Alvaro L; Wypych, Fernando; Ramos, Luiz P

    2003-01-01

    Steam explosion of Eucalyptus grandis has been carried out under various pretreatment conditions (200-210 degrees C, 2-5 min) after impregnation of the wood chips with 0.087 and 0.175% (w/w) H2SO4. This study, arranged as a 2(3) factorial design, indicated that pretreatment temperature is the most critical variable affecting the yield of steam-treated fractions. Pretreatment of 0.175% (w/w) H2SO4-impregnated chips at 210 degrees C for 2 min was the best condition for hemicellulose recovery (mostly as xylose) in the water soluble fraction, reaching almost 70% of the corresponding xylose theoretical yield. By contrast, lower pretreatment temperatures of 200 degrees C were enough to yield steam-treated substrates from which a 90% cellulose conversion was obtained in 48 h, using low enzyme loadings of a Celluclast 1.5 1 plus Novozym 188 mixture (Novo Nordisk). Release of water-soluble chromophores was monitored by UV spectroscopy and their concentration increased with pretreatment severity. The yield of alkali-soluble lignin increased at higher levels of acid impregnation and pretreatment temperatures. Thermoanalysis of these lignin fractions indicated a pattern of lignin fragmentation towards greater pretreatment severities but lignin condensation prevailed at the most drastic pretreatment conditions.

  16. Fractionation of equine antivenom using caprylic acid precipitation in combination with cationic ion-exchange chromatography.

    PubMed

    Raweerith, Rutai; Ratanabanangkoon, Kavi

    2003-11-01

    A combined process of caprylic acid (CA) precipitation and ion-exchange chromatography on SP-Sepharose was studied as a means to fractionate pepsin-digested horse antivenom F(ab')(2) antibody. In the CA precipitation, the optimal concentration for fractionation of F(ab')(2) from pepsin-digested horse plasma was 2%, in which 89.61% of F(ab')(2) antibody activity was recovered in the supernatant with 1.5-fold purification. A significant amount of pepsin was not precipitated and remained active under these conditions. An analytical cation exchanger Protein-Pak SP 8HR HPLC column was tested to establish optimal conditions for the effective separation of IgG, albumin, pepsin and CA from the F(ab')(2) product. From these results, the supernatant from CA precipitation of pepsin-digested plasma was subjected to a SP-Sepharose column chromatography using a linear salt gradient. With stepwise elution, a peak containing F(ab')(2) antibody could be obtained by elution with 0.25 M NaCl. The total recovery of antibody was 65.56% with 2.91-fold purification, which was higher than that achieved by ammonium sulfate precipitation. This process simultaneously and effectively removed residual pepsin, high molecular weight aggregates and CA in the final F(ab')(2) product, and should be suitable for large-scale fractionation of therapeutic equine antivenoms.

  17. Fluorescent properties of low-molecular-weight fractions from chernozem humic acids

    NASA Astrophysics Data System (ADS)

    Trubetskoi, O. A.; Demin, D. V.; Trubetskaya, O. E.

    2013-10-01

    The polyacrylamide gel electrophoresis of chernozem humic acids (HAs) followed by ultraviolet detection (λ = 312 nm) has revealed a new highly fluorescent fraction that has the highest electrophoretic mobility and the lowest nominal molecular weight (NMW). The preparative isolation of the fraction has been performed using the multiple microfiltration of the same HA sample in a 7 M carbamide solution on a membrane with a nominal pore size of 5 kDa. Thirty ultrafiltrates with NMW < 5 kDa and different fluorescence maximums in the region of 475-505 nm have been prepared, as well as a nonfluorescent concentrate with NMW > 5 kDa. Fluorescence maximums at and below 490 nm have been noted only in the first four ultrafiltrates. All the ultrafiltrates have been combined into the fraction with NMW < 5 kDa, which has been successively passed through membranes of 3 and 1 kDa. Solutions of subfractions F 3-5 kDa, F 1-3 kDa, and F < 1 kDa with fluorescence maximums at 505, 488, and 465 nm, respectively, have been prepared. The F < 1 kDa subfraction with the lowest NMW had the highest fluorescence intensity. The distribution of the fluorescence maximums in the ultrafiltrates has indicated the presence of at least two groups of fluorophores and has confirmed the supramolecular organization of the extracted soil HAs.

  18. Increased responsiveness to 5-hydroxytryptamine after antigenic challenge is inhibited by nifedipine and niflumic acid in rat trachea in vitro.

    PubMed

    Moura, Carlos Tiago Martins; Bezerra, Fernanda Carvalho; de Moraes, Isabelle Maciel; Magalhães, Pedro Jorge Caldas; Capaz, Francisco Ruy

    2005-12-01

    Antigenic challenge often induces hyperreactivity in asthmatic airway, although the precise mechanism(s) underlying this increased responsiveness is not entirely known. Tracheae obtained from ovalbumin (OVA)-sensitized saline- or OVA-challenged rats were placed in 10 mL bath chambers for isometric recording of 5-hydroxytryptamine (5-HT)-induced contractions. 5-Hydroxytryptamine induced a stronger contraction compared with control in antigen-challenged trachea under normal or Ca2+-free conditions. In tracheae pretreated with the L-type Ca2+ channel blocker nifedipine (10(-6) mol/L) or the Ca2+-activated Cl- channel blocker niflumic acid (10(-4) mol/L), this hyperresponsiveness was not developed in either normal or Ca2+-free medium. The increased contractile response to 5-HT in allergic rat isolated trachea may be related to a greater ionic (Ca2+ and Cl-) channel involvement.

  19. Neuroprotective effect of rough aster butanol fraction against oxidative stress in the brain of mice challenged with kainic acid.

    PubMed

    Sok, Dai-Eun; Oh, Sang Hee; Kim, Yun-Bae; Kim, Mee Ree

    2003-07-30

    The neuroprotective effect of the butanol fraction from the methanol extract of Aster scaber Thunb. (rough aster butanol fraction) on oxidative damage in the brain of mice challenged with kainic acid was examined using behavioral signs and biochemical parameters of oxidative stress. The rough aster butanol fraction (0.4-1.0 g/kg) was administered to ICR male mice, 6-8 weeks, through a gavage for 4 days consecutively, and on the third day, kainic acid (50 mg/kg) was ip administered. When compared to the vehicle-treated control, no significant changes in body and brain weight were observed in mice administered the rough aster butanol fraction. Administration of kainic acid only, causing a lethality of approximately 54%, resulted in a significant decrease of total glutathione level and an increase of the thiobarbituric acid-reactive substances (TBARS) value in brain tissue. When the rough aster butanol fraction was examined for neuroprotective action, the rough aster butanol fraction (0.4 g/kg) alleviated the lethality (25%) of kainic acid and the behavioral sign of its neurotoxicity. Moreover, administration of the rough aster butanol fraction at a dose of 0.4 g/kg restored the glutathione level in the cytosolic portion of brain homogenate to approximately 80% (p < 0.05). Also, the rough aster butanol fraction (0.4 g/kg) led to a significant reduction of kainic acid-induced increase of TBARS value. In addition, the glutathione peroxidase activity was restored significantly (p < 0.05) in the cytosolic portion of brain homogenate, whereas glutathione reductase activity was not. On the basis of these results, the rough aster butanol fraction is suggested to contain a functional agent to prevent oxidative stress in the brain of mice.

  20. Amino acid composition of some Amaranthus sp. grain proteins and of its fractions.

    PubMed

    Correa, A D; Jokl, L; Carlsson, R

    1986-09-01

    This study was carried out to determine the protein content of several Amaranthus sp. grains. Findings revealed this has a high lysine (5.3 to 6.3 of the protein) and sulphur amino acids content (3.4-4.0%), while leucine could well be limiting when those seeds are used as a sole protein source in food. Using the correction for in vitro protein digestibility, the chemical score varied from 50 to 67. The calculated protein efficiency ratios and biological values ranged from 1.39 to 1.80 and 53 to 68, respectively. Considering that amaranth grain is a good supplement to cereal grain, the protein of A. hypochondriacus HH5 (yellow seeds) and A. anclancalius (black seeds) was fractionated into albumin, globulin, prolamin and glutelin. The average proportions between those soluble proteins were 65:17:11:7, respectively. Albumin had the highest lysine content (7.3-8.2%), and globulin the highest methionine (4.1-5.3%) and phenylalanine (6.0-6.1%) content. Prolamin had the highest threonine (4.6-5.4%) and leucine (6.8-6.9%) content, while glutelin had a very low methionine content (0.6-1.0%). Based on the above-mentioned findings, the authors conclude the variation in the amino acid composition of the protein fractions can be used for genetic protein improvement.

  1. Melissa officinalis Acidic Fraction Protects Cultured Cerebellar Granule Neurons Against Beta Amyloid-Induced Apoptosis and Oxidative Stress

    PubMed Central

    Soodi, Maliheh; Dashti, Abolfazl; Hajimehdipoor, Homa; Akbari, Shole; Ataei, Nasim

    2017-01-01

    Objective Extracellular deposition of the beta-amyloid (Aβ) peptide, which is the main finding in the pathophysiology of Alzheimer’s disease (AD), leads to oxidative damage and apoptosis in neurons. Melissa officinalis (M. officinalis) is a medicinal plant from the Lamiaceae family that has neuroprotective activity. In the present study we have investigated the protective effect of the acidic fraction of M. officinalis on Aβ-induced oxidative stress and apoptosis in cultured cerebellar granule neurons (CGN). Additionally, we investigated a possible role of the nicotinic receptor. Materials and Methods This study was an in vitro experimental study performed on mice cultured CGNs. CGNs were pre-incubated with different concentrations of the acidic fraction of M. officinalis for 24 hours, followed by incubation with Aβ for an additional 48 hours. CGNs were also pre-incubated with the acidic fraction of M. officinalis and mecamylamin, followed by incubation with Aβ. We used the 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay to measure cell viability. Acetylcholinesterase (AChE) activity, reactive oxygen species (ROS) production, lipidperoxidation, and caspase-3 activity were measured after incubation. Hochst/annexin Vfluorescein isothiocyanate (FITC)/propidium iodide (PI) staining was performed to detect apoptotic cells. Results The acidic fraction could protect CGNs from Aβ-induced cytotoxicity. Mecamylamine did not abolish the protective effect of the acidic fraction. AChE activity, ROS production, lipid peroxidation, and caspase-3 activity increased after Aβ incubation. Preincubation with the acidic fraction of M. officinalis ameliorated these factors and decreased the number of apoptotic cells. Conclusion Our results indicated that the protective effect of the acidic fraction of M. officinalis was not mediated through nicotinic receptors. This fraction could protect CGNs through antioxidant and anti-apoptotic activities. PMID

  2. Identification of Candida albicans antigens reactive with immunoglobulin E antibody of human sera.

    PubMed Central

    Ishiguro, A; Homma, M; Torii, S; Tanaka, K

    1992-01-01

    Candida albicans antigens which reacted with immunoglobulin E (IgE) antibodies of 57 allergic patients were detected by immunoblotting. Of the various antigens, the 175-, 125-, 46-, 43-, and 37-kDa antigenic components reacted most frequently with the patient sera. To purify the major antigens, C. albicans cells were fractionated. The 46-, 43-, and 37-kDa antigens were recovered in cytoplasmic fractions, but the 175- and 125-kDa antigens were not recovered in any fraction. The 46-, 43-, and 37-kDa antigens were purified from cytoplasmic fractions by DEAE and P11 ion-exchange chromatography. Antigens were isolated by cutting bands out of sodium dodecyl sulfate-polyacrylamide gels. The purified components confirmed by immunoblotting were next processed for amino acid sequencing. Parts of the sequences of the 46-, 43-, and 37-kDa antigens had significant levels of homology with Saccharomyces cerevisiae glycolytic enzyme enolase, phosphoglycerate kinase, and aldolase, respectively. Rabbit IgG antibodies prepared against the 46- and 43-kDa antigens strongly cross-reacted with the homologous proteins of S. cerevisiae. However, S. cerevisiae enolase and phosphoglycerate kinase did not cross-react with IgE of patient sera. This result suggests that IgE antibodies against only small parts of their epitopes are elevated in the allergic patients. Since enolase is reported to be a major antigen for systemic candidiasis, this enzyme may be the immunodominant protein in both allergies and fungal infections. Images PMID:1548078

  3. Chemical composition and bioactivity properties of size-fractions separated from a vermicompost humic acid.

    PubMed

    Canellas, Luciano P; Piccolo, Alessandro; Dobbss, Leonardo B; Spaccini, Riccardo; Olivares, Fábio L; Zandonadi, Daniel B; Façanha, Arnoldo R

    2010-01-01

    Preparative high performance size-exclusion chromatography (HPSEC) was applied to humic acids (HA) extracted from vermicompost in order to separate humic matter of different molecular dimension and evaluate the relationship between chemical properties of size-fractions (SF) and their effects on plant root growth. Molecular dimensions of components in humic SF was further achieved by diffusion-ordered nuclear magnetic resonance spectroscopy (DOSY-NMR) based on diffusion coefficients (D), while carbon distribution was evaluated by solid state (CP/MAS) (13)C NMR. Seedlings of maize and Arabidopsis were treated with different concentrations of SF to evaluate root growth. Six different SF were obtained and their carbohydrate-like content and alkyl chain length decreased with decreasing molecular size. Progressive reduction of aromatic carbon was also observed with decreasing molecular size of separated fractions. Diffusion-ordered spectroscopy (DOSY) spectra showed that SF were composed of complex mixtures of aliphatic, aromatic and carbohydrates constituents that could be separated on the basis of their diffusion. All SF promoted root growth in Arabidopsis and maize seedlings but the effects differed according to molecular size and plant species. In Arabidopsis seedlings, the bulk HA and its SF revealed a classical large auxin-like exogenous response, i.e.: shortened the principal root axis and induced lateral roots, while the effects in maize corresponded to low auxin-like levels, as suggested by enhanced principal axis length and induction of lateral roots. The reduction of humic heterogeneity obtained in HPSEC separated size-fractions suggested that their physiological influence on root growth and architecture was less an effect of their size than their content of specific bioactive molecules. However, these molecules may be dynamically released from humic superstructures and exert their bioactivity when weaker is the humic conformational stability as that obtained

  4. Combined Dilute Acid and Solvent Based Pretreatment of Agricultural Wastes for Efficient Lignocellulosic Fractionation and Biofuels Production

    SciTech Connect

    Brodeur, G.; Ramakrishnan, S.; Wilson, C.; Telotte, J.; Collier, J.; Stickel, J.

    2013-01-01

    A true biorefinery for processing lignocellulosic biomass should achieve maximum utilization of all major constituents (cellulose, hemicellulose, & lignin) within the feedstock. In this work a combined pretreatment process of dilute acid (DA) and N-methyl morpholine N-oxide (NMMO) is described that allows for both fractionation and subsequent complete hydrolysis of the feedstocks (corn stover and sugarcane bagasse). During this multi-step processing, the dilute acid pretreatment solubilizes the majority (>90%) of the hemicellulosic fraction, while the NMMO treatment yields a cellulosic fraction that is completely digestible within 48 hours at low enzyme loadings. With both the cellulosic and hemicellulosic fractions being converted into separate, dissolved sugar fractions, the remaining portion is nearly pure lignin. When used independently, DA and NMMO pretreatments are only able to achieve ~80% and ~45% cellulosic conversion, respectively. Mass balance calculations along with experimental results are used to illustrate the feasibility of separation and recycling of NMMO.

  5. Investigating organic molecules responsible of auxin-like activity of humic acid fraction extracted from vermicompost.

    PubMed

    Scaglia, Barbara; Nunes, Ramom Rachide; Rezende, Maria Olímpia Oliveira; Tambone, Fulvia; Adani, Fabrizio

    2016-08-15

    This work studied the auxin-like activity of humic acids (HA) obtained from vermicomposts produced using leather wastes plus cattle dung at different maturation stages (fresh, stable and mature). Bioassays were performed by testing HA concentrations in the range of 100-6000mgcarbonL(-1). (13)C CPMAS-NMR and GC-MS instrumental methods were used to assess the effect of biological processes and starting organic mixtures on HA composition. Not all HAs showed IAA-like activity and in general, IAA-like activity increased with the length of the vermicomposting process. The presence of leather wastes was not necessary to produce the auxin-like activity of HA, since HA extracted from a mix of cattle manure and sawdust, where no leather waste was added, showed IAA-like activity as well. CPMAS (13)CNMR revealed that HAs were similar independently of the mix used and that the humification process involved the increasing concentration of pre-existing alkali soluble fractions in the biomass. GC/MS allowed the identification of the molecules involved in IAA-like effects: carboxylic acids and amino acids. The concentration of active molecules, rather than their simple presence in HA, determined the bio-stimulating effect, and a good linear regression between auxin-like activity and active stimulating molecules concentration was found (R(2)=-0.85; p<0.01, n=6).

  6. Modular nucleic acid assembled p/MHC microarrays for multiplexed sorting of antigen-specific T cells.

    PubMed

    Kwong, Gabriel A; Radu, Caius G; Hwang, Kiwook; Shu, Chengyi J; Ma, Chao; Koya, Richard C; Comin-Anduix, Begonya; Hadrup, Sine Reker; Bailey, Ryan C; Witte, Owen N; Schumacher, Ton N; Ribas, Antoni; Heath, James R

    2009-07-22

    The human immune system consists of a large number of T cells capable of recognizing and responding to antigens derived from various sources. The development of peptide-major histocompatibility (p/MHC) tetrameric complexes has enabled the direct detection of these antigen-specific T cells. With the goal of increasing throughput and multiplexing of T cell detection, protein microarrays spotted with defined p/MHC complexes have been reported, but studies have been limited due to the inherent instability and reproducibility of arrays produced via conventional spotted methods. Herein, we report on a platform for the detection of antigen-specific T cells on glass substrates that offers significant advantages over existing surface-bound schemes. In this approach, called "Nucleic Acid Cell Sorting (NACS)", single-stranded DNA oligomers conjugated site-specifically to p/MHC tetramers are employed to immobilize p/MHC tetramers via hybridization to a complementary-printed substrate. Fully assembled p/MHC arrays are used to detect and enumerate T cells captured from cellular suspensions, including primary human T cells collected from cancer patients. NACS arrays outperform conventional spotted arrays assessed in key criteria such as repeatability and homogeneity. The versatility of employing DNA sequences for cell sorting is exploited to enable the programmed, selective release of target populations of immobilized T cells with restriction endonucleases for downstream analysis. Because of the performance, facile and modular assembly of p/MHC tetramer arrays, NACS holds promise as a versatile platform for multiplexed T cell detection.

  7. Characterization of metabolic pathway of linoleic acid 9-hydroperoxide in cytosolic fraction of potato tubers and identification of reaction products.

    PubMed

    Kimura, Hideto; Yokota, Kazushige

    2004-01-01

    Potato tubers are shown to contain a unique lipoxygenase pathway to form 9-hydroperoxy-10,12-octadecadienoic acid (9-HPODE) from linoleic acid. Here, we report the metabolic pathway of 9-HPODE in the cytosolic fraction and the characterization of enzymes involved in the conversion of metabolites. The analysis of enzymatic reaction products at pH 5.5 revealed the formation of 9-keto-10,12-octadecadienoic acid, 9-hydroxy-10,12-octadecadienoic acid, 9,10-epoxy-11-hydroxy-12-octadecenoic acid, 9,10,13-trihydroxy-11-octadecenoic acid, and 9,12,13-trihydroxy-10-octadecenoic acid. The cytosolic enzymes were separated by anion-exchange chromatography into two fractions E1 and E2, having molecular masses of 66 and 54 kDa, respectively. The enzyme fraction E1 only produced 9-keto-10,12-octadecadienoic acid, whereas E2 formed other products. The enzyme E1 showed higher reactivity with 13- and 9-hydroperoxide of alpha-linolenic acid than 9-HPODE, but no reaction with hydroxy fatty acids. In contrast, the enzyme E2 showed the highest reactivity with 9-HPODE, followed by hydroperoxides of alpha-linolenic acid and arachidonic acid. We also evaluated the antibacterial activity of hydroxy fatty acids against Erwinia carotovora T-29, a bacterium infecting potato tubers. Growth of the bacteria was suppressed more potently with 9- or 13-hydroxy fatty acids than dihydroxy or trihydroxy fatty acids, suggesting a role for the metabolites in the resistance of bacterial infection.

  8. Antidepressant-like effects of fractions, essential oil, carnosol and betulinic acid isolated from Rosmarinus officinalis L.

    PubMed

    Machado, Daniele G; Cunha, Mauricio P; Neis, Vivian B; Balen, Grasiela O; Colla, André; Bettio, Luis E B; Oliveira, Agatha; Pazini, Francis Leonardo; Dalmarco, Juliana B; Simionatto, Edésio Luiz; Pizzolatti, Moacir G; Rodrigues, Ana Lúcia S

    2013-01-15

    The aim of this study was to investigate the antidepressant-like effect of fractions from Rosmarinus officinalis L.: ethyl acetate 1 and 2 (AcOEt1 and 2), hexane (HEX), ethanolic (ET), and essential oil-free (EOF) fractions, as well as essential oil, the isolated compounds carnosol and betulinic acid in the tail suspension test, a predictive test of antidepressant activity. Swiss mice were acutely administered by oral route (p.o.) with fractions, essential oil or isolated compounds, 60 min before the tail suspension test or open-field test. All of them produced a significant antidepressant-like effect: AcOEt1, ET, EOF fractions and essential oil (0.1-100mg/kg, p.o); HEX (0.1-10mg/kg, p.o) and AcOEt2 fraction (0.1-1mg/kg, p.o), carnosol (0.01-0.1mg/kg, p.o.) isolated from the HEX fraction and betulinic acid (10mg/kg, p.o.), isolated from the AcOEt1 and AcOEt2 fractions. No psychostimulant effect was shown in the open-field test, indicating that the effects in the tail suspension test are specific. This study suggests that carnosol and betulinic acid could be responsible for the anti-immobility effect of extracts from R. officinalis.

  9. Estimate of serum immunoglobulin G concentration using refractometry with or without caprylic acid fractionation.

    PubMed

    Morrill, K M; Polo, J; Lago, A; Campbell, J; Quigley, J; Tyler, H

    2013-07-01

    Objectives of this study were to develop a rapid calf-side test to determine serum IgG concentrations using caprylic acid (CA) fractionation, followed by refractometry of the IgG-rich supernatant and compare the accuracy of this method with results obtained using refractometry using raw serum. Serum samples (n=200) were obtained from 1-d-old calves, frozen (-20°C), and shipped to the laboratory. Samples were allowed to thaw for 1h at room temperature. Fractionation with CA was conducted by adding 1mL of serum to a tube containing 45, 60, or 75µL of CA and 0.5, 1.0, or 1.5mL of 0.06 M acetic acid. The tube contents were mixed well, allowed to react for 1 min, and then centrifuged at 3,300 × g for 0, 10, or 20 min at 25°C. The %Brix and refractive index of the fractionated supernatant were determined using a digital refractometer. Nonfractionated serum was analyzed for %Brix (BRn), refractive index (nDn), and IgG concentration by radial immunodiffusion. The mean serum IgG concentration was 19.0 mg/mL [standard deviation (SD)=9.7], with a range of 3.5 to 47.0 mg/mL. The mean serum BRn was 8.6 (SD=0.91), with a range of 6.8 to 11.0. The mean serum nDn was 1.34566 (SD=0.00140), with a range of 1.34300 to 1.34930. Serum nDn was positively correlated with IgG concentration (correlation coefficient=0.86; n=185). Fractionated samples treated with 1mL 0.6 M acetic acid and 60µL of CA and not centrifuged before analysis resulted in a strong relationship between the refractive index of the fractionated supernatant and IgG (correlation coefficient=0.80; n=45). Regression was used to determine cut points indicative of 10, 12, and 14 mg of IgG/mL to determine the sensitivity and specificity of refractometry to identify failure of passive transfer (serum IgG <10 mg/mL at 24 h old). The nDn were 1.34414, 1.34448, and 1.34480 to predict 10, 12, and 14 mg of IgG/mL of serum, respectively. The BRn cut points were 7.6, 7.8, and 8.0, respectively. The nDn cut points of 1.34448 and

  10. In vitro assessment of endocrine disrupting potential of naphthenic Acid fractions derived from oil sands-influenced water.

    PubMed

    Leclair, Liane A; Pohler, Lani; Wiseman, Steve B; He, Yuhe; Arens, Collin J; Giesy, John P; Scully, Stephen; Wagner, Brian D; van den Heuvel, Michael R; Hogan, Natacha S

    2015-05-05

    Oil sands-influenced process waters have been observed to cause reproductive effects and to induced CYP1A activity in fishes; however, little progress has been made in determining causative agents. Naphthenic acids (NAs) are the predominant organic compounds in process-affected waters, but due to the complexity of the mixture, it has been difficult to examine causal linkages in fishes. The aim of this study was to use in vitro assays specific to reproductive and CYP1A mechanisms to determine if specific acid extractable fractions of NAs obtained from oil sands-influenced water are active toward reproductive processes or interact with the Ah receptor responsible for CYP1A activity. NAs were extracted from aged oil sands-influenced waters by use of acid precipitation, and the mixture was fractionated into three acidic and one neutral fraction. The four fractions were examined for Ah receptor-mediated potency by use of the H4IIE-luc bioassay, effects on production of steroid hormones by use of the H295R steroidogenesis assay, and sex steroid receptor binding activity using the yeast estrogen screen and yeast androgen screen. The mixtures were characterized by high resolution mass spectrometry, (1)H nuclear magnetic resonance, and attenuated total reflectance infrared spectroscopy. The neutral fraction elicited Ah-receptor mediated activity after 24 h but not after 48 or 72 h. None of the fractions contained measurable levels of estrogen or androgen receptor agonists nor did they cause reductions in steroidogenesis. A number of fractions showed antiestrogenic or antiandrogenicity potency, with the neutral and main acidic fractions being the most potent. Neutral aromatic compounds are likely responsible for the CYP1A activity observed. Direct estrogenic, androgenic, or steroidogenic mechanisms are unlikely for NAs based on these results, but NAs act as potent antiandrogen or antiestrogens.

  11. Comparison between therapeutic antitoxin F(ab)2 fractionated with ammonium sulfate and caprylic acid.

    PubMed

    Redwan, El-Rashdy M

    2006-01-01

    To date, animal derived therapeutic antibodies represent the best and only choice source of antitoxins, especially in developing countries. Furthermore, this industry needs to develop a production protocol to achieve safer products. Recently, several laboratories changed their production protocol from ammonium sulfate (AS) protocol to caprylic acid (CA) fractionation. Our results showed that using the CA protocol leads to improvement in the product quality, as assessed by the albumin and protein content decrease (from 4.75 to 3.54 g/dL and 0.64 to 0.18 g/dL, respectively), which yielded a purer antitoxin product. The F(ab)2 protein aggregate formation and turbidity have been significantly reduced, 4.60 versus 2.55 and 0.046 versus 0.021 (p < 0.01), respectively. However, the anti-complementary activity was also reduced, from 42 to 33. The total IgG content was higher in CA fractionated products than AS materials. The endotoxin content was worrisome in some F(ab)2 products.

  12. Isolation and fractionation of soil humin using alkaline urea and dimethylsulphoxide plus sulphuric acid.

    PubMed

    Song, Guixue; Hayes, Michael H B; Novotny, Etelvino H; Simpson, Andre J

    2011-01-01

    Humin, the most recalcitrant and abundant organic fraction of soils and of sediments, is a significant contributor to the stable carbon pool in soils and is important for the global carbon budget. It has significant resistance to transformations by microorganisms. Based on the classical operational definition, humin can include any humic-type substance that is not soluble in water at any pH. We demonstrate in this study how sequential exhaustive extractions with 0.1 M sodium hydroxide (NaOH) + 6 M urea, followed by dimethylsulphoxide (DMSO) + 6% (v/v) sulphuric acid (H(2)SO(4)) solvent systems, can extract 70-80% of the residual materials remaining after prior exhaustive extractions in neutral and aqueous basic media. Solid-state (13)C NMR spectra have shown that the components isolated in the base + urea system were compositionally similar to the humic and fulvic acid fractions isolated at pH 12.6 in the aqueous media. The NMR spectra indicated that the major components isolated in the DMSO + H(2)SO(4) medium had aliphatic hydrocarbon associated with carboxyl functionalities and with lesser amounts of carbohydrate and peptide and minor amounts of lignin-derived components. The major components will have significant contributions from long-chain fatty acids, waxes, to cuticular materials. The isolates in the DMSO + H(2)SO(4) medium were compositionally similar to the organic components that resisted solvation and remained associated with the soil clays. It is concluded that the base + urea system released humic and fulvic acids held by hydrogen bonding or by entrapment within the humin matrix. The recalcitrant humin materials extracted in DMSO + H(2)SO(4) are largely biological molecules (from plants and the soil microbial population) that are likely to be protected from degradation by their hydrophobic moieties and by sorption on the soil clays. Thus, the major components of humin do not satisfy the classical definitions for humic

  13. Serum uric acid is associated with cardiac diastolic dysfunction among women with preserved ejection fraction.

    PubMed

    Nogi, Shinpei; Fujita, Shu-Ichi; Okamoto, Yusuke; Kizawa, Shun; Morita, Hideaki; Ito, Takahide; Sakane, Kazushi; Sohmiya, Koichi; Hoshiga, Masaaki; Ishizaka, Nobukazu

    2015-09-01

    Serum uric acid (SUA) is associated with the severity and prognosis of systolic heart failure. We investigated the potential association between SUA and cardiac diastolic dysfunction among total of 744 cardiac patients (202 women and 542 men) who had preserved left ventricular ejection fraction. Presence of diastolic dysfunction was assessed by echocardiographic data, plasma B-type natriuretic peptide concentration, and left ventricular hypertrophy. Univariate analysis showed that the prevalence of diastolic dysfunction increased with increasing SUA value in women, but not in men. When sex-nonspecific SUA quartiles were used, multivariate logistic regression analysis, among female patients who were not taking uric acid lowering medication, showed that the third (SUA, 5.7-6.4 mg) and the fourth (SUA, ≥6.5 mg/dl) SUA quartiles were associated with diastolic dysfunction with an odds ratio of 3.25 (P < 0.05) and 8.06 (P < 0.001), respectively, when compared with the first SUA quartile (≤4.7 mg/dl). When sex-specific SUA quartiles were used among these population, multivariate logistic regression analysis showed that the fourth SUA quartile (≥5.7 mg/dl) was associated with diastolic dysfunction with an odds ratio of 5.34 (P < 0.05) when compared with the first SUA quartile (≤4.1 mg/dl). By contrast, the relationship between SUA and diastolic dysfunction was not significant in men, irrespective of which of the sex-nonspecific or sex-specific SUA quartiles were used. These data indicated that among cardiac patients with preserved ejection fraction, SUA was significantly associated with diastolic dysfunction in women but not in men.

  14. Accumulation of different sulfur fractions in Chinese forest soil under acid deposition.

    PubMed

    Wang, Zhanyi; Zhang, Xiaoshan; Zhang, Yi; Wang, Zhangwei; Mulder, Jan

    2011-09-01

    Atmogenic sulfur (S) deposition loading by acid rain is one of the biggest environmental problems in China. It is important to know the accumulated S stored in soil, because eventually the size (and also the "desorption" rate) determines how rapidly the soil water pH responds to decrease in S deposition. The S fractions and the ratio of total carbon/total sulfur (C/S) of forest soil in 9 catchments were investigated by comparing soils at the rural and urban sites in China. The S fractions included water-soluble sulfate-S (SO(4)-S), adsorbed SO(4)-S, insoluble SO(4)-S and organic S. The ratio of C/S in soil at the rural site was significantly (p < 0.05) greater than that at the urban site. C/S of soil in the A horizon was significantly (p < 0.05) and negatively correlated with the wet S-deposition rate. The ratio of C/S presents a better indicator for atmogenic S loading. Organic S was the dominant form in soils at rural sites; contributing more than 69% of the total S in the uppermost 30 cm soil. Organic S and adsorbed SO(4)-S were the main forms of S in soil at urban sites. High contents of water-soluble SO(4)-S and adsorbed SO(4)-S were found in uppermost 30 cm soils at urban sites but not at rural sites. Decades of acid rain have caused accumulation of inorganic SO(4)-S in Chinese forest soil especially at the urban sites. The soil at urban sites had been firstly acidified, and the impacts on the forest ecosystem in these areas should be noticed.

  15. Transport of Corilagin, Gallic Acid, and Ellagic Acid from Fructus Phyllanthi Tannin Fraction in Caco-2 Cell Monolayers

    PubMed Central

    Zhao, Hai-juan; Liang, Wen-Yi; Chen, Wen-Jing; Han, Shu-Xian; Qi, Qi; Cui, Ya-Ping; Li, Shi; Yang, Guang-Hui; Shao, Yan-Yan; Zhu, Dan

    2016-01-01

    Objective. To investigate the absorption property of the representative hydrolyzable tannin, namely corilagin, and its hydrolysates gallic acid (GA) and ellagic acid (EA) from the Fructus Phyllanthi tannin fraction (PTF) in vitro. Methods. Caco-2 cells monolayer model was established. Influences of PTF on Caco-2 cells viability were detected with MTT assay. The transport across monolayers was examined for different time points, concentrations, and secretory directions. The inhibitors of P-glycoprotein (P-gp), multidrug resistance proteins (MRPs), organic anion transporting polypeptide (OATP) and sodium/glucose cotransporter 1 (SGLT1), and tight junction modulators were used to study the transport mechanism. LC-MS method was employed to quantify the absorption concentration. Results. The apparent permeability coefficient (Papp) values of the three compounds were below 1.0 × 10−6 cm/s. The absorption of corilagin and GA were much lower than their efflux, and the uptake of both compounds was increased in the presence of inhibitors of P-gp and MRPs. The absorption of EA was decreased in the company of OATP and SGLT1 inhibitors. Moreover, the transport of corilagin, GA, and EA was enhanced by tight junction modulators. Conclusion. These observations indicated that the three compounds in PTF were transported via passive diffusion combined with protein mediated transport. P-gp and MRPs might get involved in the transport of corilagin and GA. The absorption of EA could be attributed to OATP and SGLT1 protein. PMID:27738446

  16. Chemical Composition, Nitrogen Fractions and Amino Acids Profile of Milk from Different Animal Species

    PubMed Central

    Rafiq, Saima; Huma, Nuzhat; Pasha, Imran; Sameen, Aysha; Mukhtar, Omer; Khan, Muhammad Issa

    2016-01-01

    Milk composition is an imperative aspect which influences the quality of dairy products. The objective of study was to compare the chemical composition, nitrogen fractions and amino acids profile of milk from buffalo, cow, sheep, goat, and camel. Sheep milk was found to be highest in fat (6.82%±0.04%), solid-not-fat (11.24%±0.02%), total solids (18.05%±0.05%), protein (5.15%±0.06%) and casein (3.87%±0.04%) contents followed by buffalo milk. Maximum whey proteins were observed in camel milk (0.80%±0.03%), buffalo (0.68%±0.02%) and sheep (0.66%±0.02%) milk. The non-protein-nitrogen contents varied from 0.33% to 0.62% among different milk species. The highest r-values were recorded for correlations between crude protein and casein in buffalo (r = 0.82), cow (r = 0.88), sheep (r = 0.86) and goat milk (r = 0.98). The caseins and whey proteins were also positively correlated with true proteins in all milk species. A favorable balance of branched-chain amino acids; leucine, isoleucine, and valine were found both in casein and whey proteins. Leucine content was highest in cow (108±2.3 mg/g), camel (96±2.2 mg/g) and buffalo (90±2.4 mg/g) milk caseins. Maximum concentrations of isoleucine, phenylalanine, and histidine were noticed in goat milk caseins. Glutamic acid and proline were dominant among non-essential amino acids. Conclusively, current exploration is important for milk processors to design nutritious and consistent quality end products. PMID:26954163

  17. Immunoaffinity isolation and partial characterization of the Coccidioides immitis antigen detected by the tube precipitin and immunodiffusion-tube precipitin tests.

    PubMed Central

    Zimmer, B L; Pappagianis, D

    1989-01-01

    The antigen participating in the tube precipitin (TP) serologic test for coccidioidomycosis was isolated from mycelial-phase antigen (coccidioidin) by immunoaffinity and characterized by various analytical procedures. This was accomplished by first preparing the antigen-antibody precipitate by using antigen and human serum positive for TP (immunoglobulin M) antibody and then liberating the antigen by digestion with pronase. This protease destroyed the antibody and left the antigen intact as indicated by immunodiffusion-TP. The coccidioidal antigen was isolated from the proteolytic digest by using size exclusion chromatography. DEAE chromatography of this antigen yielded two fractions with immunodiffusion-TP reactivity which had average molecular sizes of 225 and 140 kilodaltons, respectively. The presence of carbohydrate and amino acids indicated that the antigen(s) is a glycopeptide. Compositional analysis showed that one fraction contained 3-O-methylmannose, mannose, and glucose in a ratio of 8:1.2:1, whereas the second fraction contained 3-O-methylmannose, mannose, glucose, and galactose in a ratio of 1:1:1:1. The amino acids glycine, alanine, serine, threonine, aspartic acid plus asparagine, and glutamic acid plus glutamine constituted 60 to 70% of the amino acids in both glycopeptides. Neither antigen could be detected entering the gel in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Lectin affinity provided evidence of a high-mannose asparagine-linked glycopeptide in the first peak and an asparagine-linked glycopeptide with a biantennary complex-type structure in the second peak. Images PMID:2504775

  18. Radium-226 contents and Rn emanation coefficients of particle-size fractions of alkaline, acid and mixed U mill tailings.

    PubMed

    Landa, E R

    1987-03-01

    Alkaline circuit and mixed, acid and alkaline circuit U mill tailings sampled at an inactive mill site near Monticello, UT, and tailings from an active, acid-leach U mill were separated into particle-size fractions ranging from +10 mesh to -325 mesh by dry and wet separation techniques. The 226Ra contents and 222Rn emanation coefficients of these fractions were determined. Dry tailings show a high degree of aggregation that tends to mask the relation of properties, such as Ra content and Rn emanating power, to dispersed-particle size. Coarse-tailings fractions (+325 mesh) had emanation coefficients which were from 25 to 45% lower than those of their fine-fraction counterparts. Emanation coefficients measured for tailings derived from a salt roast/carbonate-leach process suggest that such are roasting does not lead to reductions in Rn emanation in the tailings derived therefrom.

  19. Heavily fractionated noble gases in an acid residue from the Klein Glacier 98300 EH3 chondrite

    NASA Astrophysics Data System (ADS)

    Nakashima, Daisuke; Ott, Ulrich; El Goresy, Ahmed; Nakamura, Tomoki

    2010-09-01

    Noble gases were measured both in bulk samples (stepped pyrolysis and total extraction) and in a HF/HCl residue (stepped pyrolysis and combustion) from the Klein Glacier (KLE) 98300 EH3 chondrite. Like the bulk meteorite and as seen in previous studies of bulk type 3 E chondrites ("sub-Q"), the acid residue contains elementally fractionated primordial noble gases. As we show here, isotopically these are like those in phase-Q of primitive meteorites, but elementally they are heavily fractionated relative to these. The observed noble gases are different from "normal" Q noble gases also with respect to release patterns, which are similar to those of Ar-rich noble gases in anhydrous carbonaceous chondrites and unequilibrated ordinary chondrites (with also similar isotopic compositions). While we cannot completely rule out a role for parent body processes such as thermal and shock metamorphism (including a later thermal event) in creating the fractionated elemental compositions, parent body processes in general seem not be able to account for the distinct release patterns from those of normal Q noble gases. The fractionated gases may have originated from ion implantation from a nebular plasma as has been suggested for other types of primordial noble gases, including Q, Ar-rich, and ureilite noble gases. With solar starting composition, the corresponding effective electron temperature is about 5000 K. This is lower than inferred for other primordial noble gases (10,000-6000 K). Thus, if ion implantation from a solar composition reservoir was a common process for the acquisition of primordial gas, electron temperatures in the early solar system must have varied spatially or temporally between 10,000 and 5000 K. Neon and xenon isotopic ratios of the residue suggest the presence of presolar silicon carbide and diamond in abundances lower than in the Qingzhen EH3 and Indarch EH4 chondrites. Parent body processes including thermal and shock metamorphism and a late thermal

  20. Effects of olive oil and its fractions on oxidative stress and the liver's fatty acid composition in 2,4-Dichlorophenoxyacetic acid-treated rats

    PubMed Central

    2010-01-01

    Background Olive oil's beneficial effects are not only related to its high content of oleic acid, but also to the antioxidant potential of its polyphenols. In this study, we assess the effects of virgin olive oil and its fractions on 2,4-D- induced oxidative damage in the liver of rats. Methods Male Wistar rats were randomly divided into eight groups of ten each: (C) a control group, (D) group that received 2,4-D (5 mg/kg b.w.), (D/EVOO) group treated with 2,4-D plus extra virgin olive oil, (D/OOHF) group that received 2,4-D plus hydrophilic fraction, (D/OOLF) group treated with 2,4-D plus lipophilic fraction, (EVOO) group that received only extra virgin olive oil, (OOHF) group given hydrophilic fraction and (OOLF) group treated with lipophilic fraction. These components were daily administered by gavage for 4 weeks. Results A significant liver damage was observed in rats treated with 2,4-D via increased serum levels of transaminases and alkaline phosphatase, hepatic lipid peroxidation and decreased hepatic antioxidant enzyme activities, namely, superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase. The liver's fatty acid composition was also significantly modified with 2,4-D exposure. However, extra virgin olive oil and hydrophilic fraction intake during 2,4-D treatment induced a significant increase in the antioxidant enzyme activities and a decrease in the conjugated dienes (CD) and thiobarbituric acid-reactive substances (TBARs) levels in the liver. The lipophilic fraction supplemented to 2,4-D- treated rats did not show any improvement in the liver oxidative status while a marked improvement was detected in the hepatic fatty acid composition of rats supplemented with olive oil and the two fractions. Conclusion We concluded that the protective effect of olive oil against oxidative damage induced by 2,4-D is mainly related to the antioxidant potential of its hydrophilic fraction. PMID:21034436

  1. Application of a new combined fractionation technique (CFT) to detect fluorophores in size-fractionated hydrophobic acid of DOM as indicators of urban pollution.

    PubMed

    Wei, Qunshan; Yan, Changzhou; Luo, Zhuanxi; Zhang, Xian; Xu, Qiujin; Chow, Christopher W K

    2012-08-01

    For pollution monitoring, we developed a new combined fractionation technique (CFT) to characterize dissolved organic matter (DOM) in natural water by combining resin adsorption (RA) and ultrafiltration (UF) with 3D-fluorescence measurement. We tested the new technique on 4 polluted and 4 unpolluted samples. The 3D-fluorescence characteristics of size sub-components in the hydrophobic acid (HPOA) fraction could distinguish unpolluted from polluted DOM. The unpolluted HPOA fraction was composed of a single dominant size component-peak A (fulvic-like, around Ex240/Em410 nm) material with relatively large molecular weight (MW) (>10 kDa). In comparison, the HPOA in polluted DOM contained another predominant size component with lower MW (<5 kDa)-peak T material (tryptophan-like protein, around Ex230/Em340 nm). The fluorescence of peak T material with lower MW (<5 kDa) in HPOA would be a good indicator of pollution or deterioration of source water quality. The application of this new CFT could yield more detailed and scientific information on the size and chemical character of the fluorophores in DOM sub-fractions.

  2. The use of TLC-DPPH* test with image processing to study direct antioxidant activity of phenolic acid fractions of selected Lamiaceae family species.

    PubMed

    Cieśla, Łukasz; Staszek, Dorota; Kowalska, Teresa; Waksmundzka-Hajnos, Monika

    2013-01-01

    TLC coupled with 2,2-diphenyl-1-picrylhydrazyl staining was used to analyze phenolic acid fractions of selected Salvia and Thymus species. Documented videoscans were processed by means of an image processing program. This is the first time that free phenolic acids fractions, as well as fractions containing phenolic acids derived from basic and acidic hydrolysis, have been analyzed and compared for selected sage and thyme species. The analyzed samples along with caffeic acid (CA; standard) were chromatographed on silica gel plates with toluene-ethyl acetate-formic acid (60 + 40 + 1, v/v/v) mobile phase. The extracts were investigated with respect to the activity of CA. It was found that CA was most abundant in the fractions derived from basic hydrolysis. This compound was not detected in any of the fractions obtained after acidic hydrolysis. S. officinalis and S. triloba have similar free radical scavenging activity fingerprints obtained for all the analyzed fractions.

  3. Evaluation of soluble fraction and enzymatic residual fraction of dilute dry acid, ethylenediamine, and steam explosion pretreated corn stover on the enzymatic hydrolysis of cellulose.

    PubMed

    Qin, Lei; Liu, Li; Li, Wen-Chao; Zhu, Jia-Qing; Li, Bing-Zhi; Yuan, Ying-Jin

    2016-06-01

    This study is aimed to examine the inhibition of soluble fraction (SF) and enzymatic residual fraction (ERF) in dry dilute acid (DDA), ethylenediamine (EDA) and steam explosion (SE) pretreated corn stover (CS) on the enzymatic digestibility of cellulose. SF of DDA, EDA and SE pretreated CS has high xylose, soluble lignin and xylo-oligomer content, respectively. SF of EDA pretreated CS leads to the highest inhibition, followed by SE and DDA pretreated CS. Inhibition of ERF of DDA and SE pretreated CS is higher than that of EDA pretreated CS. The inhibition degree (A0/A) of SF is 1.76 and 1.21 times to that of ERF for EDA and SE pretreated CS, respectively. The inhibition degree of ERF is 1.05 times to that of SF in DDA pretreated CS. The quantitative analysis shows that SF of EDA pretreated CS, SF and ERF of SE pretreated CS cause significant inhibition during enzymatic hydrolysis.

  4. Fractionation of Carbon Isotopes in Biosynthesis of Fatty Acids by A Piezophilic Bacterium Moritella Japonica DSK1

    NASA Astrophysics Data System (ADS)

    Fang, J.; Uhle, M.; Bartlett, D.; Kato, C.

    2005-12-01

    We examined stable carbon isotope fractionation in biosynthesis of fatty acids of a piezophilic bacterium Moritella japonica DSK1. DSK1 was grown to stationary phase at pressures of 0.1, 10, 20, and 50 MPa in media prepared using natural seawater supplied with glucose with the sole carbon source. DSk1 synthesized typical bacterial fatty acids (C14-19 saturated, monounsaturated, and cyclopropane fatty acids) as well as long-chain polyunsaturated fatty acids (PUFA) (20:6ω3). Bacterial cell biomass and individual fatty acids exhibited consistent pressure-dependent carbon isotope fractionations relative to glucose. The observed ΔδFA-glucose (-1.0 to -11.9%) at 0.1 MPa was comparable to or slightly higher than fractionations reported on surface bacteria. However, Bulk biomass and fatty acids became more depleted in 13C with pressure. Average carbon isotope fractionation ΔδFA-glucose) at high pressures was much higher than that for surface bacteria: -15.7, -15.3, and -18.3‰ at 10, 20, and 50 MPa, respectively. PUFA were more 13C depleted than saturated and monounsaturated fatty acids at all pressures. The observed isotope effects may be ascribed to the kinetics of enzymatic reactions affected by hydrostatic pressure and to different biosynthetic pathways for short-chain and long-chain fatty acids. Our results have important implications for marine biogeochemistry. The 13C depleted fatty acids in marine sediments and water column may be derived simply from piezophilic bacteria resynthesis of organic matter, not from bacterial utilization of a 13C-depleted carbon source (i.e., methane). The interpretation of carbon isotope signatures of marine lipids must be based on principles derived from piezophilic bacteria.

  5. Psoralen Inactivation of Viruses: A Process for the Safe Manipulation of Viral Antigen and Nucleic Acid.

    PubMed

    Schneider, Katherine; Wronka-Edwards, Loni; Leggett-Embrey, Melissa; Walker, Eric; Sun, Peifang; Ondov, Brian; Wyman, Travis H; Rosovitz, M J; Bohn, Sherry S; Burans, James; Kochel, Tadeusz

    2015-11-12

    High consequence human pathogenic viruses must be handled at biosafety level 2, 3 or 4 and must be rendered non-infectious before they can be utilized for molecular or immunological applications at lower biosafety levels. Here we evaluate psoralen-inactivated Arena-, Bunya-, Corona-, Filo-, Flavi- and Orthomyxoviruses for their suitability as antigen in immunological processes and as template for reverse transcription PCR and sequencing. The method of virus inactivation using a psoralen molecule appears to have broad applicability to RNA viruses and to leave both the particle and RNA of the treated virus intact, while rendering the virus non-infectious.

  6. Psoralen Inactivation of Viruses: A Process for the Safe Manipulation of Viral Antigen and Nucleic Acid

    PubMed Central

    Schneider, Katherine; Wronka-Edwards, Loni; Leggett-Embrey, Melissa; Walker, Eric; Sun, Peifang; Ondov, Brian; Wyman, Travis H.; Rosovitz, MJ; Bohn, Sherry S.; Burans, James; Kochel, Tadeusz

    2015-01-01

    High consequence human pathogenic viruses must be handled at biosafety level 2, 3 or 4 and must be rendered non-infectious before they can be utilized for molecular or immunological applications at lower biosafety levels. Here we evaluate psoralen-inactivated Arena-, Bunya-, Corona-, Filo-, Flavi- and Orthomyxoviruses for their suitability as antigen in immunological processes and as template for reverse transcription PCR and sequencing. The method of virus inactivation using a psoralen molecule appears to have broad applicability to RNA viruses and to leave both the particle and RNA of the treated virus intact, while rendering the virus non-infectious. PMID:26569291

  7. Prostate-specific antigen kinetics following hypofractionated stereotactic body radiotherapy boost as post-external beam radiotherapy versus conventionally fractionated external beam radiotherapy for localized prostate cancer

    PubMed Central

    Phak, Jeong Hoon; Kim, Hun Jung; Kim, Woo Chul

    2015-01-01

    Background Stereotactic body radiotherapy (SBRT) has emerged as an effective treatment for localized prostate cancer. The purpose of this study was to compare the prostate-specific antigen (PSA) kinetics between conventionally fractionated external beam radiotherapy (CF-EBRT) and SBRT boost after whole pelvis EBRT (WP-EBRT) in localized prostate cancer. Methods A total of 77 patients with localized prostate cancer [T-stage, T1–T3; Gleason score (GS) 5–9; PSA < 20 ng/mL] were enrolled. A total of 35 patients were treated with SBRT boost (21 Gy in 3 fractions) after WP-EBRT and 42 patients were treated with CF-EBRT (45 Gy WP-EBRT and boost of 25.2–30.6 Gy in 1.8-Gy fractions). PSA nadir and rate of change in PSA (slope) were calculated and compared. Results With a median follow-up of 52.4 months (range, 14–74 months), the median PSA nadir and slope for SBRT boost were 0.29 ng/mL and −0.506, −0.235, −0.129, and −0.092 ng/mL/mo, respectively, for durations of 1 year, 2 years, 3 years, and 4 years postradiotherapy. Similarly, for CF-EBRT, the median PSA nadir and slopes were 0.39 ng/mL and −0.720 ng/mL/mo, −0.204 ng/mL/mo, −0.121 ng/mL/mo, and −0.067 ng/mL/mo, respectively. The slope of CF-EBRT was significantly different with a greater median rate of change for 1 year postradiotherapy than that of SBRT boost (P = 0.018). Contrastively, the slopes of SBRT boost for durations of 2 years, 3 years, and 4 years tended to be continuously greater than that of CF-EBRT. The significantly lower PSA nadir was observed in SBRT boost (median nadir 0.29 ng/mL) compared with CF-EBRT (median nadir 0.35 ng/mL, P = 0.025). Five-year biochemical failure (BCF) free survival was 94.3% for SBRT boost and 78.6% for CF-EBRT (P = 0.012). Conclusion Patients treated with SBRT boost after WP-EBRT experienced a lower PSA nadir and there tended to be a continuously greater rate of decline of PSA for durations of 2 years, 3 years, and

  8. Deriving Gene Networks from SNP Associated with Triacylglycerol and Phospholipid Fatty Acid Fractions from Ribeyes of Angus Cattle

    PubMed Central

    Buchanan, Justin W.; Reecy, James M.; Garrick, Dorian J.; Duan, Qing; Beitz, Don C.; Koltes, James E.; Saatchi, Mahdi; Koesterke, Lars; Mateescu, Raluca G.

    2016-01-01

    The fatty acid profile of beef is a complex trait that can benefit from gene-interaction network analysis to understand relationships among loci that contribute to phenotypic variation. Phenotypic measures of fatty acid profile from triacylglycerol and phospholipid fractions of longissimus muscle, pedigree information, and Illumina 54 k bovine SNP genotypes were utilized to derive an annotated gene network associated with fatty acid composition in 1,833 Angus beef cattle. The Bayes-B statistical model was utilized to perform a genome wide association study to estimate associations between 54 k SNP genotypes and 39 individual fatty acid phenotypes within each fraction. Posterior means of the effects were estimated for each of the 54 k SNP and for the collective effects of all the SNP in every 1-Mb genomic window in terms of the proportion of genetic variance explained by the window. Windows that explained the largest proportions of genetic variance for individual lipids were found in the triacylglycerol fraction. There was almost no overlap in the genomic regions explaining variance between the triacylglycerol and phospholipid fractions. Partial correlations were used to identify correlated regions of the genome for the set of largest 1 Mb windows that explained up to 35% genetic variation in either fatty acid fraction. SNP were allocated to windows based on the bovine UMD3.1 assembly. Gene network clusters were generated utilizing a partial correlation and information theory algorithm. Results were used in conjunction with network scoring and visualization software to analyze correlated SNP across 39 fatty acid phenotypes to identify SNP of significance. Significant pathways implicated in fatty acid metabolism through GO term enrichment analysis included homeostasis of number of cells, homeostatic process, coenzyme/cofactor activity, and immunoglobulin. These results suggest different metabolic pathways regulate the development of different types of lipids found in

  9. Influence of multi-step washing using Na2EDTA, oxalic acid and phosphoric acid on metal fractionation and spectroscopy characteristics from contaminated soil.

    PubMed

    Wei, Meng; Chen, Jiajun

    2016-11-01

    A multi-step soil washing test using a typical chelating agent (Na2EDTA), organic acid (oxalic acid), and inorganic weak acid (phosphoric acid) was conducted to remediate soil contaminated with heavy metals near an arsenic mining area. The aim of the test was to improve the heavy metal removal efficiency and investigate its influence on metal fractionation and the spectroscopy characteristics of contaminated soil. The results indicated that the orders of the multi-step washing were critical for the removal efficiencies of the metal fractions, bioavailability, and potential mobility due to the different dissolution levels of mineral fractions and the inter-transformation of metal fractions by XRD and FT-IR spectral analyses. The optimal soil washing options were identified as the Na2EDTA-phosphoric-oxalic acid (EPO) and phosphoric-oxalic acid-Na2EDTA (POE) sequences because of their high removal efficiencies (approximately 45 % for arsenic and 88 % for cadmium) and the minimal harmful effects that were determined by the mobility and bioavailability of the remaining heavy metals based on the metal stability (I R ) and modified redistribution index ([Formula: see text]).

  10. [Study of the fatty acid components of the triglyceride fraction of the blood in normal and thalassemic subjects, using gas chromatography].

    PubMed

    Gilli, G; Moiraghi Ruggenini, A; Nani, E; Bottura, G; Mastretta, L

    1977-01-01

    Thin layer chromatography was used to separate the triglyceridic fraction of plasma lipides in normal (19) and thalassaemic (15) subjects. Gas chromatographic analysis of the fraction was then carried out and the fatty acids represented were identified qualitatively and quantitatively. Statistically significant variations, specifically increase in arachidonic acid and reduction in palmitic and linoleic acids, were observed in the thalassaemic patients.

  11. Construction and application of a Korean reference panel for imputing classical alleles and amino acids of human leukocyte antigen genes.

    PubMed

    Kim, Kwangwoo; Bang, So-Young; Lee, Hye-Soon; Bae, Sang-Cheol

    2014-01-01

    Genetic variations of human leukocyte antigen (HLA) genes within the major histocompatibility complex (MHC) locus are strongly associated with disease susceptibility and prognosis for many diseases, including many autoimmune diseases. In this study, we developed a Korean HLA reference panel for imputing classical alleles and amino acid residues of several HLA genes. An HLA reference panel has potential for use in identifying and fine-mapping disease associations with the MHC locus in East Asian populations, including Koreans. A total of 413 unrelated Korean subjects were analyzed for single nucleotide polymorphisms (SNPs) at the MHC locus and six HLA genes, including HLA-A, -B, -C, -DRB1, -DPB1, and -DQB1. The HLA reference panel was constructed by phasing the 5,858 MHC SNPs, 233 classical HLA alleles, and 1,387 amino acid residue markers from 1,025 amino acid positions as binary variables. The imputation accuracy of the HLA reference panel was assessed by measuring concordance rates between imputed and genotyped alleles of the HLA genes from a subset of the study subjects and East Asian HapMap individuals. Average concordance rates were 95.6% and 91.1% at 2-digit and 4-digit allele resolutions, respectively. The imputation accuracy was minimally affected by SNP density of a test dataset for imputation. In conclusion, the Korean HLA reference panel we developed was highly suitable for imputing HLA alleles and amino acids from MHC SNPs in East Asians, including Koreans.

  12. In vitro Activities of Pfaffia glomerata Root Extract, Its Hydrolyzed Fractions and Pfaffic Acid Against Trypanosoma cruzi Trypomastigotes.

    PubMed

    Silva, Márcio L A E; Pereira, Ana C; Ferreira, Daniele S; Esperandim, Viviane R; Símaro, Guilherme V; Lima, Thaís C; Januário, Ana H; Pauletti, Patrícia M; Rehder, Vera L G; Crevelin, Eduardo J; Cunha, Wilson R; Crotti, Antônio E M; Bastos, Jairo K

    2017-01-01

    This article reports on the in vitro activity of the hydroalcoholic extract of Pfaffia glomerata roots, its hydrolyzed fractions, and pfaffic acid against Trypanosoma cruzi. The hydroalcoholic extract obtained from dried, milled P. glomerata roots was submitted to acid hydrolysis followed by partition with CHCl3 . The concentrated CHCl3 fraction was suspended in MeOH/H2 O and partitioned with hexane (F1), CHCl3 (F2), and AcOEt (F3), in this sequence. The trypanocidal activity of the hydrolyzed extract and its fractions was evaluated in vitro. The hydroalcoholic extract displayed low activity, but fraction F1 was active against trypomastigotes of the Y strain of T. cruzi, with IC50 = 47.89 μg/ml. The steroids campesterol (7.7%), stigmasterol (18.7%), β-sitosterol (16.8%), Δ(7) -stigmastenol (4.6%), and Δ(7) -spinasterol (7.5%) were the major constituents of F1, along with fatty acid esters (7.6%) and eight aliphatic hydrocarbons (30.1%). Fractions F2 and F3 exhibited moderate activity, and pfaffic acid, one of the main chemical constituents of these fractions, displayed IC50 = 44.78 μm (21.06 μg/ml). On the other hand, the hydroalcoholic extract of P. glomerata roots, which is rich in pfaffosides, was inactive. Therefore, the main aglycone of pfaffosides, pfaffic acid, is much more active against trypomastigotes of the Y strain of T. cruzi than its corresponding glycosides and should be further investigated.

  13. Vanadium and nickel complexes in petroleum resid acid, base, and neutral fractions

    SciTech Connect

    Pearson, C.D.; Green, J.D.

    1993-01-01

    Acid and base fractions from petroleum vacuum resids with no detectable (by visible spectrophotometry) quantities of porphyrinic Ni or V complexes were hydrotreated under various conditions to determine if significant amounts of porphyrinic metals were released, via disassociation or other means, upon hydrotreating. No significant quantities were observed, thereby indicating that nonporphyrinic metals were not simply associated, complexed or otherwise masked (in terms of visible spectrophotometric response) porphyrinic metal complexes. However, it is possible that hydrotreating was simply not effective in breaking up these associates and/or that some porphyrinic forms of metal were in fact released but were rapidly destroyed by hydrotreating. In addition, three liquid chromatographic (LC) separation methods were sequentially applied to Cerro Negro (Orinoco belt Venezuelan heavy crude) >700{degree}C resid in an effort to separate and concentrate the metal complexes present. Nonaqueous ion exchange chromatography was used initially to separate the resid into acid, base and neutral types. Two concentrates containing 19,500 and 13,500 ppm total V, or an estimated 19 and 13 wt % V-containing compounds respectively, were obtained. The degree of enrichment of Ni compounds obtained was significantly lower. By visible spectrophotometry, using vanadyl etioporphyrin as a standard, each of the concentrates contained near a 1:1 ratio of porphyrinic:nonporphyrinic V complexes. Analogous separation behavior for porphyrinic versus nonporphyrinic metal forms was observed throughout much of the work, thereby suggesting that a comparable diversity of structures existed within each general class of metal compounds. The generally wide dispersion of both Ni and V over the LC separation scheme suggests a structural variety of metal complexes that is comparable to that observed for other heteroatoms (N, S, O) in petroleum.

  14. Vanadium and nickel complexes in petroleum resid acid, base, and neutral fractions

    SciTech Connect

    Pearson, C.D.; Green, J.D.

    1993-01-01

    Acid and base fractions from petroleum vacuum resids with no detectable (by visible spectrophotometry) quantities of porphyrinic Ni or V complexes were hydrotreated under various conditions to determine if significant amounts of porphyrinic metals were released, via disassociation or other means, upon hydrotreating. No significant quantities were observed, thereby indicating that nonporphyrinic metals were not simply associated, complexed or otherwise masked (in terms of visible spectrophotometric response) porphyrinic metal complexes. However, it is possible that hydrotreating was simply not effective in breaking up these associates and/or that some porphyrinic forms of metal were in fact released but were rapidly destroyed by hydrotreating. In addition, three liquid chromatographic (LC) separation methods were sequentially applied to Cerro Negro (Orinoco belt Venezuelan heavy crude) >700[degree]C resid in an effort to separate and concentrate the metal complexes present. Nonaqueous ion exchange chromatography was used initially to separate the resid into acid, base and neutral types. Two concentrates containing 19,500 and 13,500 ppm total V, or an estimated 19 and 13 wt % V-containing compounds respectively, were obtained. The degree of enrichment of Ni compounds obtained was significantly lower. By visible spectrophotometry, using vanadyl etioporphyrin as a standard, each of the concentrates contained near a 1:1 ratio of porphyrinic:nonporphyrinic V complexes. Analogous separation behavior for porphyrinic versus nonporphyrinic metal forms was observed throughout much of the work, thereby suggesting that a comparable diversity of structures existed within each general class of metal compounds. The generally wide dispersion of both Ni and V over the LC separation scheme suggests a structural variety of metal complexes that is comparable to that observed for other heteroatoms (N, S, O) in petroleum.

  15. Stratospheric sulfuric acid fraction and mass estimate for the 1982 volcanic eruption of El Chichon

    NASA Technical Reports Server (NTRS)

    Hofmann, D. J.; Rosen, J. M.

    1983-01-01

    The stratospheric sulfuric acid fraction and mass for the 1982 volcanic eruptions of El Chichon are investigated using data from balloon soundings at Laramie (41 deg N) and in southern Texas (27-29 deg N). The total stratospheric mass of these eruptions is estimated to be approximately 8 Tg about 6.5 months after the eruption with possibly as much as 20 Tg in the stratosphere about 45 days after the eruption. Observations of the aerosol in Texas revealed two primary layers, both highly volatile at 150 C. Aerosol in the upper layer at about 25 km was composed of an approximately 80 percent H2SO4 solution while the lower layer at approximately 18 km was composed of a 60-65 percent H2SO4 solution aerosol. It is calculated that an H2SO4 vapor concentration of at least 3 x 10 to the 7th molecules/cu cm is needed to sustain the large droplets in the upper layer. An early bi-modal nature in the size distribution indicates droplet nucleation from the gas phase during the first 3 months, while the similarity of the large particle profiles 2 months apart shows continued particle growth 6.5 months after the explosion.

  16. Remediation of soils contaminated with chromium using citric and hydrochloric acids: the role of chromium fractionation in chromium leaching.

    PubMed

    Cheng, Shu-Fen; Huang, Chin-Yuan; Tu, Yao-Ting

    2011-01-01

    Acid washing is a common method for soil remediation, but is not always efficient for chromium-contaminated soil. Both soil particle size and the forms of chromium existing in the soil affect the efficiency of soil washing. Laboratory batch and column dissolution experiments were conducted to determine the efficiencies of citric and hydrochloric acids as agents to extract chromium from soils contaminated with chromium. The effects of soil particle size and chromium fractionation on Cr leaching were also investigated. About 90% of chromium in the studied soil existed either in residual form or bound to iron and manganese oxides, and Cr fraction distributions were similar for all soil particle sizes. Almost all exchangeable and carbonate-bound chromium was removed by washing once with 0.5 M HCl, whereas organic chromium was more effectively removed by washing with citric acid rather than with HCl solution of the same concentration. For chromium fractions that were either bound to Fe-Mn oxides or existed as residual forms, the efficiencies of acid washing were usually 20% or less, except for 0.5 M HCl solution, which had much higher efficiencies. Separation of the soil sample by particle size before the separate washing of the soil fractions had little improvement on the chromium removal.

  17. Modification of A Tumor Antigen Determinant to Improve Peptide/MHC Stability Is Associated with Increased Immunogenicity and Cross-Priming A Larger Fraction of CD8+ T Cells1

    PubMed Central

    Watson, Alan M.; Mylin, Lawrence M.; Thompson, Megan M.; Schell, Todd D.

    2012-01-01

    Altered peptide ligands (APLs) with enhanced binding to MHC class I (MHC-I) can increase the CD8+ T cell response to native antigens, including tumor antigens. Here we investigate the influence of peptide-MHC (pMHC) stability on recruitment of tumor antigen-specific CD8+ T cells through cross-priming. Among the four known H-2b-restricted CD8+ T cell determinants within SV40 large tumor antigen (TAg), the site V determinant (489QGINNLDNL497) forms relatively low-stability pMHC and is characteristically immunorecessive. Absence of detectable site V-specific CD8+ T cells following immunization with wild type TAg is due in part to inefficient cross-priming. We mutated non-anchor residues within the TAg site V determinant that increased pMHC-stability but preserved recognition by both T cell receptor transgenic and polyclonal endogenous T cells. Using a novel approach to quantify the fraction of naïve T cells triggered through cross-priming in vivo, we show that immunization with TAg variants expressing higher-stability determinants increased the fraction of site V-specific T cells cross-primed and effectively overcame the immunorecessive phenotype. In addition, using MHC-I tetramer-based enrichment we demonstrate for the first time that endogenous site V-specific T cells are primed following wild type TAg immunization despite their low initial frequency, but that the magnitude of T cell accumulation is enhanced following immunization with a site V variant TAg. Our results demonstrate that site V APLs cross-prime a higher fraction of available T cells, providing a potential mechanism for high-stability APLs to enhance immunogenicity and accumulation of T cells specific for the native determinant. PMID:23175697

  18. PSL, a nuclear cell-cycle associated antigen is increased during retinoic acid-induced differentiation of HL-60 cells.

    PubMed

    Barque, J P; Lagaye, S; Ladoux, A; Della Valle, V; Abita, J P; Larsen, C J

    1987-09-30

    PSL(p55) is a nuclear 55kD antigen present in various mammalian cell systems, which has been first identified by use of human autoimmune antibodies (Barque et al. 1983, EMBO J. 2, 743). It has been shown to be associated with interphase chromatine and to be synthesized in during the S phase of the cell cycle. In this work, we have analysed the status of PSL in promyelocytic HL-60 human cells in exponential or stationary growth, or undergoing granulocytic differentiation in presence of Retinoic acid. By use of 2-dimensional electrophoresis, PSL was found to be composed of two acidic proteins designated p55A and p55B. Unexpectedly, estimated 10-20 fold higher amounts of each species were found in cells treated for 5 days with 10(-6)M Retinoic acid, than in asynchronously growing cells or resting cells. Moreover, the p55A protein was phosphorylated during the process. On the basis of these results, PSL appears to be involved in some steps of the granulocytic differentiation process.

  19. The fraction of cells that resume growth after acetic acid addition is a strain-dependent parameter of acetic acid tolerance in Saccharomyces cerevisiae.

    PubMed

    Swinnen, Steve; Fernández-Niño, Miguel; González-Ramos, Daniel; van Maris, Antonius J A; Nevoigt, Elke

    2014-06-01

    High acetic acid tolerance of Saccharomyces cerevisiae is a relevant phenotype in industrial biotechnology when using lignocellulosic hydrolysates as feedstock. A screening of 38 S. cerevisiae strains for tolerance to acetic acid revealed considerable differences, particularly with regard to the duration of the latency phase. To understand how this phenotype is quantitatively manifested, four strains exhibiting significant differences were studied in more detail. Our data show that the duration of the latency phase is primarily determined by the fraction of cells within the population that resume growth. Only this fraction contributed to the exponential growth observed after the latency phase, while all other cells persisted in a viable but non-proliferating state. A remarkable variation in the size of the fraction was observed among the tested strains differing by several orders of magnitude. In fact, only 11 out of 10(7)  cells of the industrial bioethanol production strain Ethanol Red resumed growth after exposure to 157 mM acetic acid at pH 4.5, while this fraction was 3.6 × 10(6) (out of 10(7)  cells) in the highly acetic acid tolerant isolate ATCC 96581. These strain-specific differences are genetically determined and represent a valuable starting point to identify genetic targets for future strain improvement.

  20. Hydroxypropyl-β-cyclodextrin extractability and bioavailability of phenanthrene in humin and humic acid fractions from different soils and sediments.

    PubMed

    Gao, Huipeng; Ma, Jing; Xu, Li; Jia, Lingyun

    2014-01-01

    Organic matter (OM) plays a vital role in controlling polycyclic aromatic hydrocarbon (PAH) bioavailability in soils and sediments. In this study, both a hydroxypropyl-β-cyclodextrin (HPCD) extraction test and a biodegradation test were performed to evaluate the bioavailability of phenanthrene in seven different bulk soil/sediment samples and two OM components (humin fractions and humic acid (HA) fractions) separated from these soils/sediments. Results showed that both the extent of HPCD-extractable phenanthrene and the extent of biodegradable phenanthrene in humin fraction were lower than those in the respective HA fraction and source soil/sediment, demonstrating the limited bioavailability of phenanthrene in the humin fraction. For the source soils/sediments and the humin fractions, significant inverse relationships were observed between the sorption capacities for phenanthrene and the amounts of HPCD-extractable or biodegradable phenanthrene (p < 0.05), suggesting the importance of the sorption capacity in affecting desorption and biodegradation of phenanthrene. Strong linear relationships were observed between the amount of HPCD-extractable phenanthrene and the amount degraded in both the bulk soils/sediments and the humin fractions, with both slopes close to 1. On the other hand, in the case of phenanthrene contained in HA, a poor relationship was observed between the amount of phenanthrene extracted by HPCD and the amount degraded, with the former being much less than the latter. The results revealed the importance of humin fraction in affecting the bioavailability of phenanthrene in the bulk soils/sediments, which would deepen our understanding of the organic matter fractions in affecting desorption and biodegradation of organic pollutants and provide theoretical support for remediation and risk assessment of contaminated soils and sediments.

  1. Bioprospecting the Curculigoside-Cinnamic Acid-Rich Fraction from Molineria latifolia Rhizome as a Potential Antioxidant Therapeutic Agent.

    PubMed

    Ooi, Der Jiun; Chan, Kim Wei; Sarega, Nadarajan; Alitheen, Noorjahan Banu; Ithnin, Hairuszah; Ismail, Maznah

    2016-06-17

    Increasing evidence from both experimental and clinical studies depicts the involvement of oxidative stress in the pathogenesis of various diseases. Specifically, disruption of homeostatic redox balance in accumulated body fat mass leads to obesity-associated metabolic syndrome. Strategies for the restoration of redox balance, potentially by exploring potent plant bioactives, have thus become the focus of therapeutic intervention. The present study aimed to bioprospect the potential use of the curculigoside-cinnamic acid-rich fraction from Molineria latifolia rhizome as an antioxidant therapeutic agent. The ethyl acetate fraction (EAF) isolated from M. latifolia rhizome methanolic extract (RME) contained the highest amount of phenolic compounds, particularly curculigoside and cinnamic acid. EAF demonstrated glycation inhibitory activities in both glucose- and fructose-mediated glycation models. In addition, in vitro chemical-based and cellular-based antioxidant assays showed that EAF exhibited high antioxidant activities and a protective effect against oxidative damage in 3T3-L1 preadipocytes. Although the efficacies of individual phenolics differed depending on the structure and concentration, a correlational study revealed strong correlations between total phenolic contents and antioxidant capacities. The results concluded that enriched phenolic contents in EAF (curculigoside-cinnamic acid-rich fraction) contributed to the overall better reactivity. Our data suggest that this bioactive-rich fraction warrants therapeutic potential against oxidative stress-related disorders.

  2. Compositional and functional features of humic acid-like fractions from vermicomposting of sewage sludge and cow dung.

    PubMed

    Li, Xiaowei; Xing, Meiyan; Yang, Jian; Huang, Zhidong

    2011-01-30

    The chemical changes occurring in five different substrates of sewage sludge spiked with different proportions of cow dung after vermicomposting with Eisenia foetida for 90 days were investigated. Their humic acid-like (HAL) fractions were isolated to determine the elemental and functional composition, and structural and functional characteristics using ultraviolet/visible, Fourier transform infrared (FT-IR) and fluorescence spectroscopies and scanning electron microscopy. After vermicomposting, the total organic C and C/N ratio decreased, and the total extractable C and humic acid (HA) C increased in all substrates. In the HAL fractions, the C and H contents, C/N and C/O and aliphatic structures, proteinaceous components and carbohydrates decreased, while the O and N and acidic functional group contents and C/H ratio, aromaticity and polycondensation structures increased. Further, the results suggest that the addition of cow dung to sewage sludge could improve the quality of organic matter humification of the substrates. The structures of HAL fractions in vermicomposts resembled those typical of soil HA, especially the vermicompost of cow dung alone. Scanning electron microscopy showed the microstructure of HAL fraction in final product became close-grained and lumpy. Overall results indicate that vermicomposting was an efficient technology for promoting organic matter (OM) humification in sewage sludge and cow dung alone, as well as in mixtures of both materials, improving their quality and environmental safety as a soil OM resource for utilization as soil amendments.

  3. Molecular weight distribution of soluble fiber fractions and short chain fatty acids in ileal digesta of growing pigs.

    PubMed

    Ivarsson, E; Andersson, R; Lindberg, J E

    2012-12-01

    The effect of dietary fiber source on molecular weight (MW) distribution of soluble fiber fractions and short chain fatty acids (SCFA) in ileal digesta of 7 post valve T-cecum (PVTC) cannulated growing pigs was studied. Pigs were fed semisynthetic diets with sugar beet (Beta vulgaris) pulp (SBP) or chicory (Cichorium intybus) forage (CFO) as fiber sources of which the soluble nonstarch polysaccharide (NSP) fraction originated mainly from pectin. Three MW intervals were selected-large MW (MWL): 10,000,000 to 1,000,000 g/mol, medium MW (MWM): 1,000,000 to 200,000 g/mol, and small MW (MWS): 200,000 to 10,000 g/mol-and the relative distribution (% of total) of molecules in each interval was calculated. The MWM fraction was higher (P < 0.05) in ileal digesta of pigs fed diet SBP and the MWS fraction was higher (P < 0.05) in ileal digesta of pigs fed diet CFO. The mole/100 mole of propionic acid (HPr) was higher (P < 0.010) in pigs fed diet SBP whereas pigs fed diet CFO had higher (P < 0.010) mole/100 mole of acetic acid (HAc). The proportion of the MWL and MWM fractions in ileal digesta were negatively correlated to HAc (r = -0.52, P = 0.05, and r = -0.62, P = 0.02, respectively). The proportion of MWM in ileal digesta was positively correlated to HPr (r = 0.83; P = 0.001) whereas MWS and HPr were negatively correlated (r = -0.76; P = 0.002). In conclusion, the bacterial degradation of the soluble NSP fraction is selective and MW distribution may explain differences in SCFA production.

  4. Enantiomeric fraction determination of 2-arylpropionic acids in a package plant membrane bioreactor.

    PubMed

    Hashim, Nor H; Stuetz, Richard M; Khan, Stuart J

    2013-05-01

    Enantiomeric compositions of three 2-arylpropionic acid (2-APA) drugs, ibuprofen, naproxen, and ketoprofen, were monitored in a membrane bioreactor (MBR) treating municipal effluent in a small rural town in Australia. Specific enantiomers were determined as amide diastereomers using the chiral derivatizing reagent, (R)-1-phenylethylamine (PEA), followed by gas chromatography-tandem mass spectrometry (GC-MS/MS). The six individual enantiomers were quantified by isotope dilution and the enantiomeric fractions (EFs) were determined. Over four separate sampling events, ibuprofen EF ranged from 0.88 to 0.94 (median 0.93) in the influent and 0.38 to 0.40 (median 0.39) in the effluent. However, no significant change in ketoprofen EF was observed, with influent EFs of 0.56-0.60 (median 0.58) and effluent EFs 0.54-0.68 (median 0.56). This is the first report of enantiospecific analysis of ketoprofen in municipal wastewater and it is not yet clear why such different behavior was observed compared to ibuprofen. Naproxen EF was consistently measured at 0.99 in the influent and ranged from 0.86 to 0.94 (median 0.91) in the effluent. This study demonstrates that EF is a relatively stable parameter and does not fluctuate according to concentration or other short-term variables introduced by sampling limitations. The enantiospecific analysis of chiral chemicals presents a promising approach to elucidate a more thorough understanding of biological treatment processes and a potential tool for monitoring the performance of key biological pathways.

  5. Complex formation with nucleic acids and aptamers alters the antigenic properties of platelet factor 4

    PubMed Central

    Jaax, Miriam E.; Krauel, Krystin; Marschall, Thomas; Brandt, Sven; Gansler, Julia; Fürll, Birgitt; Appel, Bettina; Fischer, Silvia; Block, Stephan; Helm, Christiane A.; Müller, Sabine; Preissner, Klaus T.

    2013-01-01

    The tight electrostatic binding of the chemokine platelet factor 4 (PF4) to polyanions induces heparin-induced thrombocytopenia, a prothrombotic adverse drug reaction caused by immunoglobulin G directed against PF4/polyanion complexes. This study demonstrates that nucleic acids, including aptamers, also bind to PF4 and enhance PF4 binding to platelets. Systematic assessment of RNA and DNA constructs, as well as 4 aptamers of different lengths and secondary structures, revealed that increasing length and double-stranded segments of nucleic acids augment complex formation with PF4, while single nucleotides or single-stranded polyA or polyC constructs do not. Aptamers were shown by circular dichroism spectroscopy to induce structural changes in PF4 that resemble those induced by heparin. Moreover, heparin-induced anti-human–PF4/heparin antibodies cross-reacted with human PF4/nucleic acid and PF4/aptamer complexes, as shown by an enzyme immunoassay and a functional platelet activation assay. Finally, administration of PF4/44mer–DNA protein C aptamer complexes in mice induced anti–PF4/aptamer antibodies, which cross-reacted with murine PF4/heparin complexes. These data indicate that the formation of anti-PF4/heparin antibodies in postoperative patients may be augmented by PF4/nucleic acid complexes. Moreover, administration of therapeutic aptamers has the potential to induce anti-PF4/polyanion antibodies and a prothrombotic diathesis. PMID:23673861

  6. Enhanced characterization of oil sands acid-extractable organics fractions using electrospray ionization-high-resolution mass spectrometry and synchronous fluorescence spectroscopy.

    PubMed

    Bauer, Anthony E; Frank, Richard A; Headley, John V; Peru, Kerry M; Hewitt, L Mark; Dixon, D George

    2015-05-01

    The open pit oil sands mining operations north of Fort McMurray, Alberta, Canada, are accumulating tailings waste at a rate approximately equal to 4.9 million m(3) /d. Naphthenic acids are among the most toxic components within tailings to aquatic life, but structural components have largely remained unidentified. In the present study, electrospray ionization high-resolution mass spectrometry (ESI-HRMS) and synchronous fluorescence spectroscopy (SFS) were used to characterize fractions derived from the distillation of an acid-extractable organics (AEO) mixture isolated from oil sands process-affected water (OSPW). Mean molecular weights of each fraction, and their relative proportions to the whole AEO extract, were as follows: fraction 1: 237 Da, 8.3%; fraction 2: 240 Da, 23.8%; fraction 3: 257 Da, 26.7%; fraction 4: 308 Da, 18.9%; fraction 5: 355 Da, 10.0%. With increasing mean molecular weight of the AEO fractions, a concurrent increase occurred in the relative abundance of nitrogen-, sulfur-, and oxygen-containing ions, double-bond equivalents, and degree of aromaticity. Structures present in the higher-molecular-weight fractions (fraction 4 and fraction 5) suggested the presence of heteroatoms, dicarboxyl and dihydroxy groups, and organic acid compounds with the potential to function as estrogens. Because organic acid compositions become dominated by more recalcitrant, higher-molecular-weight acids during natural degradation, these findings are important in the context of oil sands tailings pond water remediation.

  7. Enhancement of propionic acid fraction in volatile fatty acids produced from sludge fermentation by the use of food waste and Propionibacterium acidipropionici.

    PubMed

    Chen, Yinguang; Li, Xiang; Zheng, Xiong; Wang, Dongbo

    2013-02-01

    Volatile fatty acids (VFA) can be used as the additional carbon source of biological nutrient removal (BNR), and the increase of propionic acid percentage in VFA has been reported to facilitate the performance of BNR. In this study a new method for significantly improving the propionic acid fraction in VFA derived from waste activated sludge was reported, which included (1) mixing food waste with sludge and pre-fermenting the mixture (first stage), and (2) separating the mixture, sterilizing the pre-fermentation liquid and fermenting it after inoculating Propionibacterium acidipropionici (second stage). By optimizing the first stage with response surface methodology, a propionic acid content of 68.4% with propionic acid concentration of 7.13 g COD/L could be reached in the second stage, which was much higher than that reported previously. Lactic acid was found to be the most abundant product of the first stage and it served as the substrate for propionic acid production in the second stage. Further investigation showed that during the first stage the addition of food waste to the pre-fermentation system of sludge significantly increased the generation of lactic acid due to the synergistic effect, which resulted in the improvement of propionic acid production in the second stage. Finally, the use of propionic acid-enriched VFA as a superior carbon source of BNR was tested, and its performance was observed to be much better than using acetic acid-enriched VFA derived from sludge by the previously documented method.

  8. Short communication: malic acid does not promote vaccenic acid accumulation in mixed ruminal fluid with fractionated fish oil by a rumen-simulation technique.

    PubMed

    Liu, L; Wang, J Q; Bu, D P; Liu, S J; Liang, S; Wei, H Y; Zhou, L Y; Liu, K L

    2008-10-01

    The objective of this study was to determine whether malic acid could promote the accumulation of vaccenic acid in the rumen. The control diet was composed of a 65:35 ratio of forage to concentrate with 1% (dry matter basis) added fractionated fish oil (rich in docosahexaenoic acid), and treatment diets consisted of the control diet with added malic acid to achieve final concentrations of 10 mM (treatment 1) and 20 mM (treatment 2), respectively. The experiment was conducted with rumen-simulation equipment (Rusitec) consisting of 9 fermenters. Each treatment included 3 fermenters as replicates. After 7 d of incubation, concentrations of vaccenic acid from treatment 1 (4.38% fatty acids) and treatment 2 (4.46% fatty acids) were similar to that of the control treatment (4.51% fatty acids). The disappearance of docosahexaenoic acid was not different among the control, treatment 1, or treatment 2. These data indicated that malic acid did not promote the accumulation of vaccenic acid in ruminal fluid.

  9. Fractionated PDT with 5-aminolevulinic acid: effective, cost effective, and patient friendly

    NASA Astrophysics Data System (ADS)

    de Vijlder, Hannah C.; Middelburg, Tom A.; de Bruijn, Henriette S.; Robinson, Dominic J.; Neumann, H. A. Martino; de Haas, Ellen R. M.

    2009-06-01

    PDT with ALA and MAL is established as a relatively effective treatment for non-melanoma skin cancer and premalignancies. PDT is often repeated, because a single treatment gives poor long term results. Preclinical studies showed that ALA-PDT applying a fractionated illumination scheme with a small first light fraction and a second larger light fraction separated by a dark interval of two hours resulted in a significant increase in efficacy. Whereas the efficacy was not enhanced by fractionating MAL-PDT, indicating that ALA-PDT mechanism is not the same as MAL-PDT mechanism. The increase in efficacy using fractionated PDT was confirmed clinically. A randomized comparative clinical study comparing fractionated ALA-PDT versus non-fractionated ALA-PDT in the treatment of superficial basal cell carcinoma showed a significant higher response rate in the lesions treated with fractionated ALA-PDT after a follow-up of one year ( p<0.002, log-rank test). The five year follow-up is studied at moment. So far the complete response in the group treated with fractionated ALA-PDT seems to be only a few percentages lower compared to the one year follow-up. Besides the gain in response rate, fractionated ALA PDT is cost effective. ALA gel is less expensive than the commercially available MAL (Metvix) and moreover fractionated ALA-PDT takes one treatment day, instead of two treatment days using the Metvix treatment protocol (two MAL-PDT treatments separated by one week), both reducing direct and indirect costs and the burden to the patient.

  10. Fractionation of carbon isotopes in biosynthesis of fatty acids by a piezophilic bacterium Moritella japonica strain DSK1

    NASA Astrophysics Data System (ADS)

    Fang, Jiasong; Uhle, Maria; Billmark, Kaycie; Bartlett, Douglas H.; Kato, Chaki

    2006-04-01

    We examined stable carbon isotope fractionation in biosynthesis of fatty acids of a piezophilic bacterium Moritella japonica strain DSK1. The bacterium was grown to stationary phase at pressures of 0.1, 10, 20, and 50 MPa in media prepared using sterile-filtered natural seawater supplied with glucose as the sole carbon source. Strain DSK1 synthesized typical bacterial fatty acids (C 14-19 saturated, monounsaturated, and cyclopropane fatty acids) as well as long-chain polyunsaturated fatty acids (PUFA) (20:6 ω3). Bacterial cell biomass and individual fatty acids exhibited consistent pressure-dependent carbon isotope fractionations relative to glucose. The observed Δδ FA-glucose (-1.0‰ to -11.9‰) at 0.1 MPa was comparable to or slightly higher than fractionations reported in surface bacteria. However, bulk biomass and fatty acids became more depleted in 13C with pressure. Average carbon isotope fractionation (Δδ FA-glucose) at high pressures was much higher than that for surface bacteria: -15.7‰, -15.3‰, and -18.3‰ at 10, 20, and 50 MPa, respectively. PUFA were more 13C depleted than saturated and monounsaturated fatty acids at all pressures. The observed isotope effects may be ascribed to the kinetics of enzymatic reactions that are affected by hydrostatic pressure and to biosynthetic pathways that are different for short-chain and long-chain fatty acids. A simple quantitative calculation suggests that in situ piezophilic bacterial contribution of polyunsaturated fatty acids to marine sediments is nearly two orders of magnitude higher than that of marine phytoplankton and that the carbon isotope imprint of piezophilic bacteria can override that of surface phytoplankton. Our results have important implications for marine biogeochemistry. Depleted fatty acids reported in marine sediments and the water column may be derived simply from piezophilic bacteria resynthesis of organic matter, not from bacterial utilization of a 13C-depleted carbon source (i

  11. Two-dimensional electrophoresis and characterization of antigens from Paracoccidioides brasiliensis.

    PubMed

    da Fonseca, C A; Jesuino, R S; Felipe, M S; Cunha, D A; Brito, W A; Soares, C M

    2001-06-01

    Paracoccidioides brasiliensis is a fungal pathogen of humans. To identify antigens from P. brasiliensis we fractionated a crude preparation of proteins from the fungus and detected the IgG reactive proteins by immunoblot assays of yeast cellular extracts with sera of patients with paracoccidioidomycosis (PCM). We identified and characterized six new antigens by amino acid sequencing and homology search analyses with other proteins deposited in a database. The newly characterized antigens were highly homologous to catalase, fructose-1,6-biphosphate aldolase (aldolase), glyceraldehyde-3-phosphate dehydrogenase, malate dehydrogenase and triosephosphate isomerase from several sources. The characterized antigens presented preferential synthesis in yeast cells, the host fungus phase.

  12. Effects of USDA beef quality grade and cooking on fatty acid composition of neutral and polar lipid fractions.

    PubMed

    Legako, J F; Dinh, T T N; Miller, M F; Brooks, J C

    2015-02-01

    The effects of USDA beef quality grade (QG; Prime, Low Choice, and Standard; n=8) and cooking (RC) on fatty acid (FA) concentrations (mg/g dry matter) and percentages of neutral and polar lipid fractions (NL and PL, respectively)from strip steaks were explored. An increase in QG led to an accumulation of most FA, especially in the NL fraction (P < 0.001). Common effects on FA percentages were two-way interactions of either QG or RC with LF (P ≤ 0.019). Fatty acids were affected differently by QG and RC depending on their originating LF. Monounsaturated fatty acid (MUFA) and polyunsaturated fatty acid (PUFA) percentages of the PL were dependent on QG (P ≤ 0.014). Cooking and QG had minimal impact on FA percentages of the NL, however, greatly influenced PL MUFA and PUFA percentages (P b 0.001). There was evidence indicating that dry heat cookery affected not only PUFA, as generally thought, but also the MUFA of PL fraction.

  13. The highly antigenic 53/25 kDa Taenia solium protein fraction with cathepsin-L like activity is present in the oncosphere/cysticercus and induces non-protective IgG antibodies in pigs.

    PubMed

    Zimic, Mirko; Pajuelo, Mónica; Gilman, Robert H; Gutiérrez, Andrés H; Rueda, Luis D; Flores, Myra; Chile, Nancy; Verástegui, Manuela; Gonzalez, Armando; García, Héctor H; Sheen, Patricia

    2012-01-15

    Cathepsin L-like proteases are secreted by several parasites including Taenia solium. The mechanism used by T. solium oncospheres to degrade and penetrate the intestine and infect the host is incompletely understood. It is assumed that intestinal degradation is driven by the proteolytic activity of enzymes secreted by the oncosphere. Blocking the proteolytic activity by an antibody response would prevent the oncosphere penetration and further infection. Serine and cysteine proteases including chymotrypsin, trypsin, elastase, and cathepsin L, are secreted by T. solium and Taenia saginata oncospheres when cultured in vitro, being potential vaccine candidates. However, the purification of a sufficient quantity of proteases secreted by oncospheres to conduct a vaccine trial is costly and lengthy. A 53/25 kDa cathepsin L-like fraction partially purified from T. solium cyst fluid was described previously as an important antigen for immunodiagnostics. In this study we found that this antigen is present in the T. solium oncosphere and is also secreted by the cysticercus. This protein fraction was tested for its ability to protect pigs against an oral challenge with T. solium oncospheres in a vaccine trial. IgG antibodies against the 53/25 kDa cathepsin L-like protein fraction were elicited in the vaccinated animals but did not confer protection.

  14. Chemical composition of fatty acid and unsaponifiable fractions of leaves, stems and roots of Arbutus unedo and in vitro antimicrobial activity of unsaponifiable extracts.

    PubMed

    Diba, Mohamed Amine; Paolini, Julien; Bendahou, Mourad; Varesi, Laurent; Allali, Hocine; Desjobert, Jean-Marie; Tabti, Boufeldja; Costa, Jean

    2010-07-01

    The chemical composition of the fatty acid and unsaponifiable fractions of the leaves, stems and roots of Arbutus unedo L. were determined using gas chromatography and gas chromatography-mass spectrometry. The fatty acid fractions of the leaves, stems and roots contained 38.5%, 31.3% and 14.1% palmitic acid, respectively, along with other long-chain fatty acids (up to C22). The chemical composition of the unsaponifiable fractions differed: the leaf and stem fractions contained high levels of aliphatic (32.1% and 62.6%, respectively) and terpenic compounds (49.6% and 25.7%, respectively), and the root fraction mainly contained esters, of which the most abundant was benzyl cinnamate (36.6%). The antimicrobial activities of the unsaponifiable fractions against nine species of microorganisms were assessed. The unsaponifiable leaf and stem extracts inhibited the growth of Klebsiella pneumoniae, Enterococcus faecalis and Candida albicans.

  15. H-binding of size- and polarity-fractionated soil and lignite humic acids after removal of metal and ash components.

    PubMed

    Drosos, Marios; Leenheer, Jerry A; Avgeropoulos, Apostolos; Deligiannakis, Yiannis

    2014-03-01

    A fractionation technique, combining dialysis removal of metal and ash components with hydrofluoric acid and pH 10 citrate buffer followed by chromatography of dialysis permeate on XAD-8 resin at decreasing pH values, has been applied to lignite humic acid (lignite-HA) and soil humic acid (soil-HA). H-binding data and non ideal competitive adsorption-Donnan model parameters were obtained for the HA fractions by theoretical analysis of H-binding data which reveal a significant increase of the carboxyl and the phenolic charge for the lignite-HA fractions vs. the parental lignite humic acid (LParentalHA). The fractionated lignite-HA material consisted mainly of permeate fractions, some of which were fulvic acid-like. The fractionated soil-HA material consisted mainly of large macromolecular structures that did not permeate the dialysis membrane during deashing. Chargeable groups had comparable concentrations in soil-HA fractions and parental soil humic acid (SParentalHA), indicating minimal interference of ash components with carboxyl and phenolic (and/or enolic) groups. Fractionation of HA, combined with theoretical analysis of H-binding, can distinguish the supramolecular vs. macromolecular nature of fractions within the same parental HA.

  16. Vi Antigen Biosynthesis in Salmonella typhi: Characterization of UDP-N-acetylglucosamine C-6 Dehydrogenase (TviB) and UDP-N-acetylglucosaminuronic Acid C-4 Epimerase (TviC)†

    PubMed Central

    Zhang, Hua; Zhou, Ying; Bao, Hongbo; Liu, Hung-wen

    2008-01-01

    Vi antigen, the virulence factor of Salmonella typhi, has been used clinically as a molecular vaccine. TviB and TviC are two enzymes involved in the formation of Vi antigen, a linear polymer consisting of α-1,4-linked N-acetylgalactosaminuronate. Protein sequence analysis suggests that TviB is a dehydrogenase and TviC is an epimerase. Both enzymes are expected to be NAD+ dependent. In order to verify their functions, TviB and TviC were cloned, expressed in Escherichia coli, and characterized. The C-terminal His6-tagged TviB protein, purified from soluble cell fractions in the presence of 10 mM DTT, shows UDP-N-acetylglucosamine 6-dehydrogenase activity, and is capable of catalyzing the conversion of UDP-N-acetylglucosamine (UDP-GlcNAc) to UDP-N-acetylglucosaminuronic acid (UDP-GlcNAcA) with a kcat value of 15.5 ± 1.0 min−1. The Km values of TviB for UDP-GlcNAc and NAD+ are 77 ± 9 μM and 276 ± 52 μM, respectively. TviC, purified as C-terminal hexahistidine-tagged protein, shows UDP-GlcNAcA 4-epimerase and UDP-N-acetylgalactosamine (UDP-GalNAc) 4-epimerase activities. The Km values of TviC for UDP-GlcNAcA and UDP-N-acetylgalactosaminuronic acid (UDP-GalNAcA) are 20 ± 1 μM and 42 ± 2 μM, respectively. The kcat value for the conversion of UDP-GlcNAcA to UDP-GalNAcA is 56.8 ± 0.5 min−1, while that for the reverse reaction is 39.1 ± 0.6 min−1. These results show that the biosynthesis of Vi antigen is initiated by the TviB-catalyzed oxidation of UDP-GlcNAc to UDP-GalNAc, followed by the TviC-catalyzed epimerization at C-4 to form UDP-GalNAcA, which serves as the building block for the formation of Vi polymer. These results set the stage for future in vitro biosynthesis of Vi antigen. These enzymes may also be drug targets to inhibit Vi antigen production. PMID:16800641

  17. Antimicrobial Effects of a Lipophilic Fraction and Kaurenoic Acid Isolated from the Root Bark Extracts of Annona senegalensis

    PubMed Central

    Okoye, Theophine Chinwuba; Akah, Peter Achunike; Okoli, Charles Ogbonnaya; Ezike, Adaobi Chioma; Omeje, Edwin Ogechukwu; Odoh, Uchenna Estella

    2012-01-01

    Root bark preparation of Annona senegalensis Pers. (Annonaceae) is used in Nigerian ethnomedicine for treatment of infectious diseases. Extraction of the A. senegalensis powdered root bark with methanol-methylene chloride (1 : 1) mixture yielded the methanol-methylene extract (MME) which was fractionated to obtain the ethyl acetate fraction (EF). The EF on further fractionation gave two active subfractions, F1 and F2. The F1 yielded a lipophilic oily liquid while F2 on purification, precipitated white crystalline compound, AS2. F1 was analyzed using GC-MS, while AS2 was characterized by proton NMR and X-ray crystallography. Antibacterial and antifungal studies were performed using agar-well-diffusion method with 0.5 McFarland standard and MICs calculated. GC-MS gave 6 major constituents: kaur-16-en-19-oic acid; 1-dodecanol; 1-naphthalenemethanol; 6,6-dimethyl-bicyclo[3.1.1]hept-2-ene-2-ethanol; 3,3-dimethyl-2-(3-methylbuta-1,3-dienyl)cyclohexane-1-methanol; 3-hydroxyandrostan-17-carboxylic acid. AS2 was found to be kaur-16-en-19-oic acid. The MICs of EF, F1, and AS2 against B. subtilis were 180, 60, and 30 μg/mL, respectively. AS2 exhibited activity against S. aureus with an MIC of 150 μg/mL, while F1 was active against P. aeruginosa with an MIC of 40 μg/mL. However, the extracts and AS2 exhibited no effects against Candida albicans and Aspergillus niger. Therefore, kaurenoic acid and the lipophilic fraction from A. senegalensis root bark exhibited potent antibacterial activity. PMID:22675389

  18. Production of high-oleic acid tallow fractions using lipase-catalyzed directed interesterification, using both batch and continuous processing.

    PubMed

    MacKenzie; Stevenson

    2000-08-01

    Immobilized lipases were used to catalyze batch-directed interesterification of tallow, resulting in oleins containing significantly higher levels of unsaturated fatty acids than obtained by fractionation without lipase. After 14 days, a reaction catalyzed by 2% Novozym 435 yielded 57% olein unsaturation, compared with 45% in a no-enzyme control. Free fatty acid levels increased to 2-3% during reactions. Incubation of the enzyme in multiple batches of melted fat caused a gradual loss of interesterification activity, apparently due to progressive dehydration. The activity could be restored by addition of water to the reaction medium. Immobilized lipase was also used to catalyze directed interesterification in a continuous flow reactor. Melted tallow was circulated through a packed bed enzyme reactor and a separate crystallization vessel. The temperatures of the two parts of the apparatus were controlled separately to allow crystallization to occur separately from interesterification. Operation of the reactor with conventionally dry, prefractionated tallow allowed the formation of an olein consisting of up to 60% unsaturated fatty acids. The greatest changes in olein fatty acid composition were achieved when the fractionation temperature was kept constant at a value that promoted selective crystallization of trisaturated triglycerides that were continuously produced by enzymic interesterification. The enzyme could be reused without apparent loss of activity, and its activity was apparently enhanced by preincubation in melted tallow for up to several days. Control of both the water activity of the enzyme and tallow feedstock and of the absorption of atmospheric water vapor were required to maintain enzyme activity, during multiple reuse and minimize free fatty acid formation. This method may form the basis for a process to produce highly mono-unsaturated tallow fractions for use in food applications (e.g. frying) where a "healthy" low saturated fat product is required.

  19. Disruption of HLA class II antigen presentation in Burkitt lymphoma: implication of a 47,000 MW acid labile protein in CD4+ T-cell recognition.

    PubMed

    God, Jason M; Zhao, Dan; Cameron, Christine A; Amria, Shereen; Bethard, Jennifer R; Haque, Azizul

    2014-07-01

    While Burkitt lymphoma (BL) has a well-known defect in HLA class I-mediated antigen presentation, the exact role of BL-associated HLA class II in generating a poor CD4(+) T-cell response remains unresolved. Here, we found that BL cells are deficient in their ability to optimally stimulate CD4(+) T cells via the HLA class II pathway. This defect in CD4(+) T-cell recognition was not associated with low levels of co-stimulatory molecules on BL cells, as addition of external co-stimulation failed to elicit CD4(+) T-cell activation by BL. Further, the defect was not caused by faulty antigen/class II interaction, because antigenic peptides bound with measurable affinity to BL-associated class II molecules. Interestingly, functional class II-peptide complexes were formed at acidic pH 5·5, which restored immune recognition. Acidic buffer (pH 5·5) eluate from BL cells contained molecules that impaired class II-mediated antigen presentation and CD4(+) T-cell recognition. Biochemical analysis showed that these molecules were greater than 30,000 molecular weight in size, and proteinaceous in nature. In addition, BL was found to have decreased expression of a 47,000 molecular weight enolase-like molecule that enhances class II-mediated antigen presentation in B cells, macrophages and dendritic cells, but not in BL cells. These findings demonstrate that BL likely has multiple defects in HLA class II-mediated antigen presentation and immune recognition, which may be exploited for future immunotherapies.

  20. Amino acid sequences recognized by T cells: studies on a merozoite surface antigen from the FCQ-27/PNG isolate of Plasmodium falciparum.

    PubMed

    Rzepczyk, C M; Csurhes, P A; Baxter, E P; Doran, T J; Irving, D O; Kere, N

    1990-08-01

    Twenty-six overlapping peptides, spanning the entire FCQ-27/PNG sequence of the Plasmodium falciparum antigen known as merozoite surface antigen 2 were screened for their ability to induce the proliferation of peripheral blood lymphocytes (PBL) obtained from 12 donors living in Honiara, Solomon Islands where P. falciparum is endemic. A recombinant (r) form of MSA2, known as Ag 1609 was also screened in these assays and tetanus toxoid (TT) antigen was included as a control. The location of the predicted T cell determinants within MSA2 was examined using the algorithm, AMPHI and by scanning MSA2 for amino acid sequences showing the Rothbard motif. There were 13 predicted amphipathic helical sites and five examples of Rothbard sequences in the antigen. The location of these with regard to the peptides tested is shown. Nine of the 12 individuals responded to TT with high stimulation indices (greater than 4) being obtained in the majority of donors. Only three individuals responded to r-MSA2 with the stimulation indices (SI) in the range of 2.4-4.1. Peptides from both the constant and variable regions of MSA2 were recognized in the proliferative assays. However, the majority of the positive proliferative responses were to peptides which spanned the central variable region which included the two copies of the 32-amino-acid repeat occurring in the antigen. High SI comparable to those obtained to TT were seen in some individuals with some peptides. There was considerable variation between donors in number and nature of the peptides recognised and two donors did not respond to any of the antigens tested. The significance of these findings to vaccine development is discussed.

  1. Identification of anti-HIV active dicaffeoylquinic- and tricaffeoylquinic acids in Helichrysum populifolium by NMR-based metabolomic guided fractionation.

    PubMed

    Heyman, Heino Martin; Senejoux, François; Seibert, Isabell; Klimkait, Thomas; Maharaj, Vinesh Jaichand; Meyer, Jacobus Johannes Marion

    2015-06-01

    South Africa being home to more than 35% of the world's Helichrysum species (c.a. 244) of which many are used in traditional medicine, is seen potentially as a significant resource in the search of new anti-HIV chemical entities. It was established that five of the 30 Helichrysum species selected for this study had significant anti-HIV activity ranging between 12 and 21 μg/mL (IC50) by using an in-house developed DeCIPhR method on a full virus model. Subsequent toxicity tests also revealed little or no toxicity for these active extracts. With the use of NMR-based metabolomics, the search for common chemical characteristics within the plant extract was conducted, which resulted in specific chemical shift areas identified that could be linked to the anti-HIV activity of the extracts. The NMR chemical shifts associated with the activity were identified to be 2.56-3.08 ppm, 5.24-6.28 ppm, 6.44-7.04 ppm and 7.24-8.04 ppm. This activity profile was then used to guide the fractionation process by narrowing down and focusing the fractionation and purification processes to speed up the putative identification of five compounds with anti-HIV activity in the most active species, Helichrysum populifolium. The anti-HIV compounds identified for the first time from H. populifolium were three dicaffeoylquinic acid derivatives, i.e. 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid and 4,5-dicaffeoylquinic acid as well as two tricaffeoylquinic acid derivatives i.e. 1,3,5-tricaffeoylquinic acid and either 5-malonyl-1,3,4-tricaffeoylquinic or 3-malonyl-1,4,5-tricaffeoylquinic acid, with the latter being identified for the first time in the genus.

  2. Development of preparative and analytical methods of the hop bitter acid oxide fraction and chemical properties of its components.

    PubMed

    Taniguchi, Yoshimasa; Matsukura, Yasuko; Taniguchi, Harumi; Koizumi, Hideki; Katayama, Mikio

    2015-01-01

    The bitter acids in hops (Humulus lupulus L.) and beer, such as α-, β-, and iso-α-acids, are known to affect beer quality and display various physiological effects. However, these compounds readily oxidize, and the effect of the oxides on the properties of beer or their potential health benefits are not well understood. In this study, we developed a simple preparative method for the bitter acid oxide fraction derived from hops and designated the constituents as matured hop bitter acids (MHBA). HPLC-PDA-ESI/HRMS and MS(2) revealed that MHBA are primarily composed of α-acid-derived oxides, which possess a common β-tricarbonyl moiety in their structures similar to α-, β-, and iso-α-acids. We also developed a quantitative analytical method of whole MHBA by HPLC, which showed high precision and reproducibility. Using our newly developed method, the concentration of whole MHBA in several commercial beers was evaluated. Our results will promote the study of bitter acid oxides.

  3. Serum sialic acid and prostate-specific antigen in differential diagnosis of benign prostate hyperplasia and prostate cancer.

    PubMed

    Romppanen, Jarkko; Haapalainen, Terhi; Punnonen, Kari; Penttilä, Ilkka

    2002-01-01

    In order to improve the diagnostic accuracy of the serum total and free prostate-specific antigen (PSA) in differential diagnosis between benign prostate hyperplasia (BPH) and prostate cancer, the serum total sialic acid (TSA) was measured and logistic regression (LR) models were built. Significantly higher serum PSA (p<0.001) concentrations were observed in patients with prostate cancer compared to control subjects, but no statistically significant differences were found in serum TSA concentrations between these groups. Serum PSA reliably discriminated patients with prostate cancer from control subjects, the area under the ROC curve (AUC) being 0.991 (0.010). When serum PSA was in the gray zone, from 4 to 10 microg/l, the diagnostic accuracy of PSA in discriminating patients with prostate cancer from BPH patients was very poor, AUC being 0.563 (0.132). However, using the same set of patients the LR model combining serum PSA, free to total PSA ratio and TSA values, as well as digital rectal examination results, had good diagnostic accuracy in discriminating the prostate cancer patients from patients with BPH, the area under the ROC curve being 0.895 (0.054). The present data suggest that the logistic regression model combining laboratory measurements and results of the clinical examination may be a useful adjunct in the differential diagnosis of benign and malignant prostate disease.

  4. Comparison of colorimetric assays with quantitative amino acid analysis for protein quantification of Generalized Modules for Membrane Antigens (GMMA).

    PubMed

    Rossi, Omar; Maggiore, Luana; Necchi, Francesca; Koeberling, Oliver; MacLennan, Calman A; Saul, Allan; Gerke, Christiane

    2015-01-01

    Genetically induced outer membrane particles from Gram-negative bacteria, called Generalized Modules for Membrane Antigens (GMMA), are being investigated as vaccines. Rapid methods are required for estimating the protein content for in-process assays during production. Since GMMA are complex biological structures containing lipid and polysaccharide as well as protein, protein determinations are not necessarily straightforward. We compared protein quantification by Bradford, Lowry, and Non-Interfering assays using bovine serum albumin (BSA) as standard with quantitative amino acid (AA) analysis, the most accurate currently available method for protein quantification. The Lowry assay has the lowest inter- and intra-assay variation and gives the best linearity between protein amount and absorbance. In all three assays, the color yield (optical density per mass of protein) of GMMA was markedly different from that of BSA with a ratio of approximately 4 for the Bradford assay, and highly variable between different GMMA; and approximately 0.7 for the Lowry and Non-Interfering assays, highlighting the need for calibrating the standard used in the colorimetric assay against GMMA quantified by AA analysis. In terms of a combination of ease, reproducibility, and proportionality of protein measurement, and comparability between samples, the Lowry assay was superior to Bradford and Non-Interfering assays for GMMA quantification.

  5. Serum tissue polypeptide antigen (TPA) and prostatic acid phosphatase (PAP) in patients with prostatic cancer.

    PubMed

    Marczyńska, A; Kulpa, J; Leńko, J; Augustyn, M

    1988-01-01

    Concurrent measurements of serum TPA and PAP concentrations by double antibody radioimmunoassays were done in 49 patients with prostatic cancer in different clinical stages. The reference group comprised patients suffering from BPH. Positive TPA was found in 32.7% of cancer patients, the lowest percentage in stage A (11.1%) and the highest in stage D (55.6%). The additional value as a diagnostic aid of the TPA test was revealed on the basis of examination of the selected group of patients with not increased PAP. Positive TPA was found in 16.7% of patients: none in stage A, 22.2% in stage B, and 33.3% in stage D. Prostatic cancer remains the most common malignancy of the genitourinary tract. The improvement in the results of treatment involves not only a modernization of treatment modalities but also the introduction of laboratory tests which give the most ample information on the stage of tumour development and improve possibilities to control tumour therapy. Besides the refinement of the determination procedures of specific prostatic markers, prostatic acid phosphatase (PAP), through radio- and enzyme-immunological methods, there is a search for additional markers which might be helpful in diagnosis and follow-up of treatment.

  6. New application of a subcellular fractionation method to kidney and testis for the determination of conjugated linoleic acid in selected cell organelles of healthy and cancerous human tissues.

    PubMed

    Hoffmann, Kristina; Blaudszun, Jörg; Brunken, Claus; Höpker, Wilhelm-Wolfgang; Tauber, Roland; Steinhart, Hans

    2005-03-01

    To clarify the mechanism of the anticarcinogenic effect of conjugated linoleic acid (CLA), its intracellular distribution needs to be determined. Subcellular fractionation using centrifugation techniques is a method that is frequently used for isolation of cell organelles from different tissues. But as the size and density of the organelles differ, the method needs to be optimised for every type of tissue. The novelty of this study is the application of a subcellular fractionation method to human healthy and cancerous renal and testicular tissue. Separation of total tissue homogenate into nuclei, cytosol, and a mixture of mitochondria and plasma membranes was achieved by differential centrifugation. As mitochondria and plasma membranes seemed to be too similar in size and weight to be separated by differential centrifugation, discontinuous density-gradient centrifugation was carried out successfully. The purity of the subcellular fractions was checked by measuring the activity of marker enzymes. All fractions were highly enriched in their corresponding marker enzyme. However, the nuclear fractions of kidney and renal cell carcinoma were slightly contaminated with mitochondria and plasma membrane fractions of all tissues with lysosomes. The fraction designated the cytosolic fraction contained not only cytosol, but also microsomes and lysosomes. The CLA contents of the subcellular fractions were in the range 0.13-0.37% of total fatty acids and were lowest in the plasma membrane fractions of all types of tissue studied. C16:0, C18:0, C18:1 c9, C18:2 n-6, and C20:4 n-6 were found to be the major fatty acids in all the subcellular fractions studied. However, marked variations in fatty acid content between subcellular fractions and between types of tissue were detectable. Because of these differences between tissues, no general statement on characteristic fatty acid profiles of single subcellular fractions is possible.

  7. Liposome-Antigen-Nucleic Acid Complexes Protect Mice from Lethal Challenge with Western and Eastern Equine Encephalitis Viruses

    PubMed Central

    Phillips, Aaron T.; Schountz, Tony; Toth, Ann M.; Rico, Amber B.; Jarvis, Donald L.; Powers, Ann M.

    2014-01-01

    Alphaviruses are mosquito-borne viruses that cause significant disease in animals and humans. Western equine encephalitis virus (WEEV) and eastern equine encephalitis virus (EEEV), two New World alphaviruses, can cause fatal encephalitis, and EEEV is a select agent of concern in biodefense. However, we have no antiviral therapies against alphaviral disease, and current vaccine strategies target only a single alphavirus species. In an effort to develop new tools for a broader response to outbreaks, we designed and tested a novel alphavirus vaccine comprised of cationic lipid nucleic acid complexes (CLNCs) and the ectodomain of WEEV E1 protein (E1ecto). Interestingly, we found that the CLNC component, alone, had therapeutic efficacy, as it increased survival of CD-1 mice following lethal WEEV infection. Immunization with the CLNC-WEEV E1ecto mixture (lipid-antigen-nucleic acid complexes [LANACs]) using a prime-boost regimen provided 100% protection in mice challenged with WEEV subcutaneously, intranasally, or via mosquito. Mice immunized with LANACs mounted a strong humoral immune response but did not produce neutralizing antibodies. Passive transfer of serum from LANAC E1ecto-immunized mice to nonimmune CD-1 mice conferred protection against WEEV challenge, indicating that antibody is sufficient for protection. In addition, the LANAC E1ecto immunization protocol significantly increased survival of mice following intranasal or subcutaneous challenge with EEEV. In summary, our LANAC formulation has therapeutic potential and is an effective vaccine strategy that offers protection against two distinct species of alphavirus irrespective of the route of infection. We discuss plausible mechanisms as well the potential utility of our LANAC formulation as a pan-alphavirus vaccine. PMID:24257615

  8. Sorption-desorption of imidacloprid onto a lacustrine Egyptian soil and its clay and humic acid fractions.

    PubMed

    Kandil, Mahrous M; El-Aswad, Ahmed F; Koskinen, William C

    2015-01-01

    Sorption-desorption of the insecticide imidacloprid 1-[(6-chloro-3-pyridinyl)-methyl]-N-nitro-2-imidazolidinimine onto a lacustrine sandy clay loam Egyptian soil and its clay and humic acid (HA) fractions was investigated in 24-h batch equilibrium experiments. Imidacloprid (IMDA) sorption-desorption isotherms onto the three sorbents were found to belong to a non-linear L-type and were best described by the Freundlich model. The value of the IMDA adsorption distribution coefficient, Kd(ads), varied according to its initial concentration and was ranged 40-84 for HA, 14-58 for clay and 1.85-4.15 for bulk soil. Freundlich sorption coefficient, Kf(ads), values were 63.0, 39.7 and 4.0 for HA, clay and bulk soil, respectively. The normalized soil Koc value for imidacloprid sorption was ∼800 indicating its slight mobility in soils. Nonlinear sorption isotherms were indicated by 1/n(ads) values <1 for all sorbents. Values of the hysteresis index (H) were <1, indicating the irreversibility of imidacloprid sorption process with all tested sorbents. Gibbs free energy (ΔG) values indicated a spontaneous and physicosorption process for IMDA and a more favorable sorption to HA than clay and soil. In conclusion, although the humic acid fraction showed the highest capacity and affinity for imidacloprid sorption, the clay fraction contributed to approximately 95% of soil-sorbed insecticide. Clay and humic acid fractions were found to be the major two factors controlling IMDA sorption in soils. The slight mobility of IMDA in soils and the hysteresis phenomenon associated with the irreversibility of its sorption onto, mainly, clay and organic matter of soils make its leachability unlikely to occur.

  9. Boric Acid Adsorption onto Humic Acids: Structures, Stabilities, 11B NMR and 11B,10B Isotopic Fractionations of Surface Complexes

    NASA Astrophysics Data System (ADS)

    Tossell, J. A.

    2006-05-01

    Boric acid, B(OH)3, forms complexes in aqueous solution with a number of bidentate O-containing ligands, L- 2, such as C2O4-2 (oxalate), C3H2O4-2 (malonate), C2H4O2-2 (glycolate), C6H4O2-2 (catecholate) and C10H6O2-2 (dioxynaphthalene). McElligot and Byrne (1998) have also found it to form a complex with CO3-2. Recently Lemarchand, et al. (2005) have studied the formation of surface complexes of B(OH)3 on humic acid, determining formation constants, 11B NMR shifts and 11B,10B isotope fractionations for a number of such complexes. This helps to clarify both the interaction of boric acid with the humic acid and the nature of the coordinating sites on the humic acid. The determination of isotope fractionations may be seen as a form of vibrational spectroscopy, using the fractionating element as a local probe of the vibrational spectrum which determines the fractionation. We have calculated quantum mechanically the structures, stabilities, vibrational spectra, 11B NMR spectra and 11B,110B isotope fractionations of a number of complexes B(OH)2L- formed by reactions of the type: B(OH)3 + HL- ? B(OH)2L- + H2O (1) using the 6-311G(d,p) B3LYP method for structures and isotopic fractionations, the highly accurate Complete Basis Set-QB3 method for energetics and the GIAO HF method with a 6-311+G(2d,p) basis for the NMR shieldings. Oxalic acid, malonic acid and catechol all form stable complexes (?G<0 for reaction 1),which are deshielded (less negative ?) vs. B(OH)4- by 2.7, 0.6 and 5.6 ppm, respectively, and which are isotopically lighter than B(OH)4- (more negative ? 11B) by 3, 2 and 5 ‰, respectively. The calculated 11B NMR shifts match well with the results of Lemarchand, et al. (2005) while the calculated isotopic fractionations are qualitatively consistent with their results, but show much smaller deviations from B(OH)4-. This is probably a consequence of the use by these authors of a value of 1.0194 (Kakihana, et al., 1977) for the 11B,10B isotopic exchange

  10. An experimental protocol for the fractionation and 2DE separation of HeLa and A-253 cell lysates suitable for the identification of the individual antigenic proteome in Sjögren's syndrome.

    PubMed

    Nginamau, Elisabeth Sivy; Maehle, Bjørn Ove; Jonsson, Roland

    2011-12-01

    Sjögren's syndrome (SS) is an autoimmune disease affecting exocrine glands, especially the salivary and lacrimal glands. Although most of the SS patients' sera have autoantibodies that can target a variety of antigens, it is not clear what determines which proteins will become autoantigens. The muscarinic receptor M3, an integral plasma membrane protein, has been proposed as a possible autoantigen in SS, and is endogenous in HeLa cells. The aim of this study was to develop a method that is able to separate and identify antigens recognised by sera from SS patients using lysates of HeLa and A-253 cells in 2D Western Blot (2DWB). The HeLa and A-253 cell lysates were fractionated in soluble and membrane-bound proteins, and the membrane-bound proteins were enriched for integral proteins. The fractions were tested using WB, confirming the presence of the main cell compartments. The rehydration solution containing ASB-14 performed better than the others in all three steps (active rehydration, focus and transfer), and efficiently separated the muscarinic receptor M3. The M3 receptor was also detected in lysates from A-253 cells. The presence of this receptor in this cell line has not been proven earlier. This work develops a suitable protocol to perform a mapping of the autoantibodies present in the sera of single SS patients, using lysates from epithelial cell lines that represent the main cell compartments as an antigen source. It is our future aim to use this protocol to perform a mapping of the antibodies present in the sera of individual SS patients.

  11. Effects of simulated acid rain, EDTA, or their combination, on migration and chemical fraction distribution of extraneous metals in Ferrosol.

    PubMed

    Wen, Fang; Hou, Hong; Yao, Na; Yan, Zengguang; Bai, Liping; Li, Fasheng

    2013-01-01

    A laboratory repacked soil-leaching column experiment was conducted to study the effects of simulated acid rain or EDTA by themselves or in combination, on migration and chemical speciation distribution of Pb and its alternative rare metals including Ag, Bi, In, Sb, and Sn. Experimental results demonstrate that leaching with simulated acid rain promoted the migration of Bi, In and Pb, and their migration reached down to 8 cm in the soil profile, no enhancement of Sb, Ag or Sn migration was observed. Addition of EDTA significantly enhanced the migration of all six metals, especially Bi, In and Pb. The migration of metals was in the order Pb>Bi>In>Sb>Sn>Ag. The individual and combined effects of acid rain and EDTA increased the environmental risk of metals, by increasing the soluble content of metals in soil solutions and the relative distribution of the exchangeable fraction. Leaching risks of Bi, In and Pb were higher than other three metals.

  12. Simultaneous determination of Ca, Cu, Ni, Zn and Cd binding strengths with fulvic acid fractions by Schubert's method

    USGS Publications Warehouse

    Brown, G.K.; MacCarthy, P.; Leenheer, J.A.

    1999-01-01

    The equilibrium binding of Ca2+, Ni2+, Cd2+, Cu2+ and Zn2+ with unfractionated Suwannee river fulvic acid (SRFA) and an enhanced metal binding subfraction of SRFA was measured using Schubert's ion-exchange method at pH 6.0 and at an ionic strength (??) of 0.1 (NaNO3). The fractionation and subfractionation were directed towards obtaining an isolate with an elevated metal binding capacity or binding strength as estimated by Cu2+ potentiometry (ISE). Fractions were obtained by stepwise eluting an XAD-8 column loaded with SRFA with water eluents of pH 1.0 to pH 12.0. Subfractions were obtained by loading the fraction eluted from XAD-8 at pH 5.0 onto a silica gel column and eluting with solvents of increasing polarity. Schuberts ion exchange method was rigorously tested by measuring simultaneously the conditional stability constants (K) of citric acid complexed with the five metals at pH 3.5 and 6.0. The logK of SRFA with Ca2+, Ni2+, Cd2+, Cu2+ and Zn2+ determined simultaneously at pH 6.0 follow the sequence of Cu2+>Cd2+>Ni2+>Zn2+>Ca2+ while all logK values increased for the enhanced metal binding subfraction and followed a different sequence of Cu2+>Cd2+>Ca2+>Ni2+>Zn2+. Both fulvic acid samples and citric acid exhibited a 1:1 metal to ligand stochiometry under the relatively low metal loading conditions used here. Quantitative 13C nuclear magnetic resonance spectroscopy showed increases in aromaticity and ketone content and decreases in aliphatic carbon for the elevated metal binding fraction while the carboxyl carbon, and elemental nitrogen, phosphorus, and sulfur content did not change. The more polar, elevated metal binding fraction did show a significant increase in molecular weight over the unfractionated SRFA. Copyright (C) 1999 Elsevier Science B.V.

  13. Amino acid composition, molecular weight distribution and gel electrophoresis of walnut (Juglans regia L.) proteins and protein fractionations.

    PubMed

    Mao, Xiaoying; Hua, Yufei; Chen, Guogang

    2014-01-27

    As a by-product of oil production, walnut proteins are considered as an additional source of plant protein for human food. To make full use of the protein resource, a comprehensive understanding of composition and characteristics of walnut proteins are required. Walnut proteins have been fractionated and characterized in this study. Amino acid composition, molecular weight distribution and gel electrophoresis of walnut proteins and protein fractionations were analyzed. The proteins were sequentially separated into four fractions according to their solubility. Glutelin was the main component of the protein extract. The content of glutelin, albumin, globulin and prolamin was about 72.06%, 7.54%, 15.67% and 4.73% respectively. Glutelin, albumin and globulin have a balanced content of essential amino acids, except for methionine, with respect to the FAO pattern recommended for adults. SDS-PAGE patterns of albumin, globulin and glutelin showed several polypeptides with molecular weights 14.4 to 66.2 kDa. The pattern of walnut proteins in two-dimension electrophoresis (2-DE) showed that the isoelectric point was mainly in the range of 4.8-6.8. The results of size exclusion chromatogram indicated molecular weight of the major components of walnut proteins were between 3.54 and 81.76 kDa.

  14. Influence of gelatinization on the extraction of phenolic acids from wheat fractions

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The effect of gelatinization on the analysis of phenolic acids from wheat bran, whole-wheat, and refined flour samples was investigated using two extraction procedures, namely, ultrasonic (UAE) and microwave (MAE). The total phenolic acid (TPA) concentration quantity in wheat bran (2711-2913 µg/g) w...

  15. [Cd uptake in rice cultivars and Cd fractions in soil treated with organic acids and EDTA].

    PubMed

    Zhang, Hai-Bo; Li, Yang-Rui; Xu, Wei-Hong; Chen, Gui-Qing; Wang, Hui-Xian; Han, Gui-Qi; Zhang, Xiao-Jing; Xiong, Zhi-Ting; Zhang, Jin-Zhong; Xie, De-Ti

    2011-09-01

    A pot experiment was conducted to examine the yield, quality and cadmium (Cd) uptake in different rice cultivars, and Cd speciation in soil after exposing to Cd (0, 1 and 5 mg x kg(-1)) in the presence of organic acids and ethylenediamine tetraacetic acid (EDTA). The results showed that general increase in the yield for cultivars Xiushui63 and II you527 was observed. Yield of two rice cultivars were in order of organic acids treatment or organic acids + 1/2EDTA treatment > EDTA treatment. The exchangeable, carbonate related and ferric-manganese oxidation related Cd increased; while organic complexation Cd and residules decreased in the presence of organic acids and EDTA. Cadmium concentrations in grain, straw and roots of both cultivars markedly reduced in the presence of organic acids and EDTA. Grain Cd concentration was the lowest for plants treated with EDTA, followed by organic acids + 1/2EDTA, and the highest Cd concentration in grain was found in the treatment with organic acids. Grain Cd concentration decreased by 9.0% to 49.3% and 16.5% to 30.6% at 1 mg x kg(-1) Cd in the presence of organic acids and EDTA, and by 12.7% to 28.5% and 4.3% to 19.1% at 5 mg x kg(-1) Cd. Cadmium concentration and accumulation in plants and total Cd content in soil were higher in Xiushui63 than in that in II you527. Grain Cd concentration decreased, and yield and quality of two rice cultivars increased at the same time in the presence of organic acids + 1/2EDTA.

  16. Selective purification of reduced oligosaccharides using a phenylboronic acid bond elut column: potential application in HPLC, mass spectrometry, reductive amination procedures and antigenic/serum analysis.

    PubMed

    Stoll, M S; Hounsell, E F

    1988-11-01

    A phenylboronic acid (PBA) derivatized Bond Elut column has been investigated for the selective purification of oligosaccharide alditols and of oligosaccharide derivatives formed by reductive amination reactions. These compounds are specifically retained by the column via interaction of sterically unhindered vicinal hydroxyl groups, hence achieving their separation from glycopeptides, unreduced oligosaccharides and non-carbohydrate reactants. This purification procedure has widespread application for the clean-up of derivatives prior to HPLC, mass spectrometry and antigenic/serum analysis.

  17. Suppression of antigen-specific antibody responses in mice exposed to perfluorooctanoic acid: Role of PPARa and T- and B-cell targeting

    EPA Pesticide Factsheets

    Dosing information, body weights during exposure and immune system endpoints. This dataset is associated with the following publication:DeWitt, J., W. Williams , J. Creech, and R. Luebke. Suppression of antigen-specific antibody responses in mice exposed to perfluorooctanoic acid: Role of PPARalpha and T- and B-cell targeting. JOURNAL OF IMMUNOTOXICOLOGY. Taylor & Francis, Inc., Philadelphia, PA, USA, 13(1): 38-45, (2016).

  18. Carbon and nitrogen isotope fractionation of amino acids in an avian marine predator, the gentoo penguin (Pygoscelis papua)

    PubMed Central

    McMahon, Kelton W; Polito, Michael J; Abel, Stephanie; McCarthy, Matthew D; Thorrold, Simon R

    2015-01-01

    Compound-specific stable isotope analysis (CSIA) of amino acids (AA) has rapidly become a powerful tool in studies of food web architecture, resource use, and biogeochemical cycling. However, applications to avian ecology have been limited because no controlled studies have examined the patterns in AA isotope fractionation in birds. We conducted a controlled CSIA feeding experiment on an avian species, the gentoo penguin (Pygoscelis papua), to examine patterns in individual AA carbon and nitrogen stable isotope fractionation between diet (D) and consumer (C) (Δ13CC-D and Δ15NC-D, respectively). We found that essential AA δ13C values and source AA δ15N values in feathers showed minimal trophic fractionation between diet and consumer, providing independent but complimentary archival proxies for primary producers and nitrogen sources respectively, at the base of food webs supporting penguins. Variations in nonessential AA Δ13CC-D values reflected differences in macromolecule sources used for biosynthesis (e.g., protein vs. lipids) and provided a metric to assess resource utilization. The avian-specific nitrogen trophic discrimination factor (TDFGlu-Phe = 3.5 ± 0.4‰) that we calculated from the difference in trophic fractionation (Δ15NC-D) of glutamic acid and phenylalanine was significantly lower than the conventional literature value of 7.6‰. Trophic positions of five species of wild penguins calculated using a multi-TDFGlu-Phe equation with the avian-specific TDFGlu-Phe value from our experiment provided estimates that were more ecologically realistic than estimates using a single TDFGlu-Phe of 7.6‰ from the previous literature. Our results provide a quantitative, mechanistic framework for the use of CSIA in nonlethal, archival feathers to study the movement and foraging ecology of avian consumers. PMID:25859333

  19. Antigens and allergens in Dermatophagoïdes farinae mite. I. Immunochemical and physicochemical study of two allergenic fractions from a partially-purified Dermatophagoïdes farinae mite extract.

    PubMed Central

    Le Mao, J; Dandeu, J P; Rabillon, J; Lux, M; David, B

    1981-01-01

    The fractionation of a partially-purified extract of Dermatophagoïdes farinae mite culture has been undertaken by gel filtration. Two fractions were isolated. One, P25, is a protein-rich fraction with mol. wt about 25,000. The other, GP8, is a polysaccharide-rich fraction with mol. wt around 8,000. By crossed immunoelectrophoresis, we detected eleven antigens in the partially-purified dialysed D. farinae extract (Df 80d) as well as in P25 fraction. One of them, ag 11, seems the most important allergen since in crossed-radio immunoelectrophoresis experiments it displays the faster radiostaining, implying that it binds the greatest part of the mite-specific IgE present in a pool of sera from mite-sensitive patients. By crossed-line immunoelectrophoresis, we demonstrated the absence of ag 11 in GP8, in which only ag 5 and ag 6 were identifiable. By radioalloergosorbent tests (RAST), it was found that P25- and GP8- coated paper discs can fix specific IgE induced in the majority of D. farinae sensitive patients. Defining a 'major allergen' as an allergen to which the majority of sensitive patients develop specific IgE, both P25 and GP8 do appear to contain at least one major allergen. By RAST inhibition method, using Df 80d as a solid phase, the allergenic activity of P25 appeared as slightly higher than that of Df 80d, whereas GP8 displayed a very weak inhibitor capacity. Thus, the allergic specificity of GP8 differs from that of Df 80d or P25. Images Figure 2 Figure 3 Figure 4 PMID:7298067

  20. Boric acid adsorption on humic acids: Ab initio calculation of structures, stabilities, 11B NMR and 11B, 10B isotopic fractionations of surface complexes

    NASA Astrophysics Data System (ADS)

    Tossell, J. A.

    2006-10-01

    Boric acid, B(OH) 3, forms complexes in aqueous solution with a number of bidentate O-containing ligands, HL -, where H 2L is C 2O 4H 2 (oxalic acid), C 3O 4H 4 (malonic acid), C 2H 6O 2 (ethylene glycol), C 6H 6O 2 (catechol), C 10H 8O 2 (dioxynaphthalene) and C 2O 3H 4 (glycolic acid). McElligott and Byrne [McElligott, S., Byrne, R.H., 1998. Interaction of B(OH)30 and HCO3- in seawater: Formation of B(OH)CO3-. Aquat. Geochem.3, 345-356.] have also found B(OH) 3 to form an aqueous complex with HCO3-1. Recently Lemarchand et al. [Lemarchand, E., Schott, J., Gaillardeet, J., 2005. Boron isotopic fractionation related to boron sorption on humic acid and the structure of surface complexes formed. Geochim. Cosmochim. Acta69, 3519-3533] have studied the formation of surface complexes of B(OH) 3 on humic acid, determining 11B NMR shifts and fitted values of formation constants, and 11B, 10B isotope fractionations for a number of surface complexation models. Their work helps to clarify both the nature of the interaction of boric acid with the functional groups in humic acid and the nature of some of these coordinating sites on the humic acid. The determination of isotope fractionations may be seen as a form of vibrational spectroscopy, using the fractionating element as a local probe of the vibrational spectrum. We have calculated quantum mechanically the structures, stabilities, vibrational spectra, 11B NMR spectra and 11B, 10B isotope fractionations of a number of complexes B(OH) 2L - formed by reactions of the type: B(OH)3+HL-⇒B(OH)2L+HO using a 6-311G(d,p) basis set and the B3LYP method for determination of structures, vibrational frequencies and isotopic fractionations, the highly accurate Complete Basis Set-QB3 method for calculating the free energies and the GIAO HF method with a 6-311+G(2d,p) basis for the NMR shieldings. The calculations indicate that oxalic acid, malonic acid, catechol and glycolic acid all form stable complexes (Δ G < 0 for Reaction (1

  1. Influences of fatty acid moiety and esterification of polyglycerol fatty acid esters on the crystallization of palm mid fraction in oil-in-water emulsion.

    PubMed

    Sakamoto, Mitsuhiro; Ohba, Azusa; Kuriyama, Juhei; Maruo, Kouichi; Ueno, Satoru; Sato, Kiyotaka

    2004-08-15

    We examined the crystallization of palm mid fraction (PMF) in oil-in-water (O/W) emulsion, after adding polyglycerol fatty acid esters (PGFEs). We employed ultrasonic velocity measurements and DSC techniques, with special emphases on the influences of fatty acid moiety and esterification of PGFE. Twelve types of PGFEs were examined as additives. PGFEs have a large hydrophilic moiety composed of 10 glycerol molecules to which palmitic, stearic and behenic acids were esterified as the fatty acid moiety with different degrees of esterification. Crystallization temperature (T(c)) of PMF remarkably increased with increasing concentrations of the PGFEs as the chain length of the fatty acid moiety increased, and the PGFE became more hydrophobic in accordance with increasing degree of esterification. We observed that the heterogeneous nucleation of PMF in the O/W emulsion was activated at the oil-water interface, where the template effect of very hydrophobic long saturated fatty acid chains of the PGFE might play the main role of heterogeneous nucleation.

  2. Phytochemical Profiling of Flavonoids, Phenolic Acids, Terpenoids, and Volatile Fraction of a Rosemary (Rosmarinus officinalis L.) Extract.

    PubMed

    Mena, Pedro; Cirlini, Martina; Tassotti, Michele; Herrlinger, Kelli A; Dall'Asta, Chiara; Del Rio, Daniele

    2016-11-19

    This paper presents a comprehensive analysis of the phytochemical profile of a proprietary rosemary (Rosmarinus officinalis L.) extract rich in carnosic acid. A characterization of the (poly)phenolic and volatile fractions of the extract was carried out using mass spectrometric techniques. The (poly)phenolic composition was assessed by ultra-high performance liquid chromatography-electrospray ionization-mass spectrometry (UHPLC-ESI-MS(n)) and a total of 57 compounds were tentatively identified and quantified, 14 of these being detected in rosemary extract for the first time. The rosemary extract contained 24 flavonoids (mainly flavones, although flavonols and flavanones were also detected), 5 phenolic acids, 24 diterpenoids (carnosic acid, carnosol, and rosmanol derivatives), 1 triterpenoid (betulinic acid), and 3 lignans (medioresinol derivatives). Carnosic acid was the predominant phenolic compound. The volatile profile of the rosemary extract was evaluated by head space solid-phase microextraction (HS-SPME) linked to gas chromatography-mass spectrometry (GC-MS). Sixty-three volatile molecules (mainly terpenes, alcohols, esters, aldehydes, and ketones) were identified. This characterization extends the current knowledge on the phytochemistry of Rosmarinus officinalis and is, to our knowledge, the broadest profiling of its secondary metabolites to date. It can assist in the authentication of rosemary extracts or rosemary-containing products or in testing its bioactivity. Moreover, this methodological approach could be applied to the study of other plant-based food ingredients.

  3. In vitro release of arachidonic acid and in vivo responses to respirable fractions of cotton dust

    SciTech Connect

    Thomson, T.A.; Edwards, J.H.; Al-Zubaidy, T.S.; Brown, R.C.; Poole, A.; Nicholls, P.J.

    1986-04-01

    It was considered that the fall in lung function seen after exposure to cotton dust may be attributable in part to the activity of arachidonic acid metabolites, such as leucotrienes as well as to the more established release of histamine by cotton dust. However, we found that cotton and barley dusts elicited poor release of arachidonic acid from an established macrophage like cell line compared with that observed with other organic dusts. In the experimental animal, pulmonary cellular responses to both cotton and barley dust were similar to those evoked by moldy hay and pigeon dropping dusts, although after multiple doses a more severe response was seen to cotton and barley. Since both moldy hay and pigeon droppings elicit a greater arachidonic acid release than cotton or barley, a role for arachidonic acid in inducing the cellular response is less likely than other factors. There are limitations to our conclusions using this system, i.e., the arachidonic acid may be released in a nonmetabolized form, although it is noted that the two dusts with the greatest arachidonic acid release produce their clinical responses in humans largely by hypersensitivity mechanisms.

  4. Effect of low-molecular-weight organic acids on kinetics release and fractionation of phosphorus in some calcareous soils of western Iran.

    PubMed

    Taghipour, M; Jalali, M

    2013-07-01

    Organic acid has been related to nutrient mobilization, mainly in phosphorus (P) insoluble utilization, and therefore enhances P bioavailability. In this study, we examined the effect of low-molecular-weight organic acids (malic, citric, and oxalic acids) on P release of some calcareous soils from western Iran. Fractionation and speciation of P in the soil solution were studied at the initial and final P release. Significantly different quantities of P were extracted by the organic acids. On average the maximum (1,554.9 mg kg(-1)) and the minimum (1,260.5 mg kg(-1)) P were extracted by 10 mM oxalic and malic acid, respectively. Power equation described well P release. In the initial stage of P release, the solution samples in soils were supersaturated with respect to hydroxyapatite and β-TCP. At the end of P release, all solutions were undersaturated with phosphate minerals. The percentage of Fe-Al oxide fraction generally increased after P release, while carbonate and residual P fractions were decreased in all organic acids. Compared with the native soils, adding malic and citric acids had no effect on Fe-Al oxide fraction, but oxalic acid significantly reduced this fraction.

  5. Distribution of carbon isotopes in amino acids of protein fraction of micro-organisms as a means of studying the mechanisms of their biosynthesis in the cell

    SciTech Connect

    Ivlev, A.A.

    1986-04-10

    The intramolecular distribution of carbon isotopes in the amino acids of the protein fraction of a number of photosynthesizing microorganisms was analyzed using the previously proposed model of carbon isotope fractionation in the cell. A correlation was found between the distributions of the isotopes in the amino acids and the pathways and sequence of their synthesis in the cell cycle. The feasibility of using the isotopic distributions of metabolites for a study of the temporal organization of metabolism in the cell is illustrated.

  6. Isotopic composition of Xe in the most acid-resistant fractions of the Efremovka CV3 chondrite

    NASA Astrophysics Data System (ADS)

    Fisenko, A. V.; Semenova, L. F.; Lavrukhina, A. K.; Shukoliukov, Iu. A.

    1991-06-01

    Isotopic compositions of three samples (E19, E20, and E22) of the Efremovka carbonaceous chondrite, obtained after 'hard' acid treatment of the original specimens were investigated. Results indicate that the fine-grained E19 contains Xe enriched by nucleogenic Xe-X. Samples E20 and E22 were found to be enriched by Xe-136, Xe-134, Xe-128, Xe-126, and Xe-124, in ratios that have no analogies among acid-resistant residues of other carbonaceous chondrites. It is suggested that the enrichment of Efremovka samples by heavy Xe isotopes was caused by the presence of Xe = H (CCF = Xe) or a mixture of extensively mass-fractionated solar Xe with Xe = X, whereas the enrichment by light Xe isotopes was caused by a intense neutron irradiation of some of the mineral phases of these samples previous to accretion.

  7. Europium(III) complexed by HPSEC size-fractions of a vertisol humic acid: small differences evidenced by time-resolved luminescence spectroscopy.

    PubMed

    Reiller, Pascal E; Brevet, Julien; Nebbioso, Antonio; Piccolo, Alessandro

    2011-03-01

    The size fractionation of a humic acid (HA) by high performance size exclusion chromatography (HPSEC) was used as a proxy for the filtration effect during HA transport through a porous medium with minimum specific chemical interactions. The modification of the Eu(III)-HA complexes' formation with the different size-fractions, as compared to the bulk HA, was studied in time-resolved luminescence spectroscopy (TRLS). Clear modifications in Eu(III)-HA complexes' structures were shown and related to the molecular characteristics of the separated size-fractions. The properties of most of size-fractions did not induce a major alteration of the affinity towards Eu(III). Only the most hydrophilic fractions eluted in the tail of the chromatographic peak, representing about 11% of total fractions-weight, gave some significantly different parameters. Using a simplistic complexation model, it was found that the available complexation sites decreased with the size reduction of humic fractions.

  8. Hypoglycemic effects of a phenolic acid fraction of rice bran and ferulic acid in C57BL/KsJ-db/db mice.

    PubMed

    Jung, Eun Hee; Kim, Sung Ran; Hwang, In Kyeong; Ha, Tae Youl

    2007-11-28

    Rice bran contains many phenolic acids, the most abundant of which is the antioxidant, ferulic acid (FA). We evaluated the hypoglycemic effects of a phenolic acid fraction (the ethyl acetate fraction, EAE) of rice bran and of FA in C57BL/KsJ db/db mice. Type 2 diabetic mice were allocated to a control group, an EAE group, or an FA group. Animals were fed a modified AIN-76 diet, and EAE or FA was administered orally for 17 days. There was no significant difference in body weight gain between groups. Administration of EAE and FA significantly decreased blood glucose levels and increased plasma insulin levels. EAE or FA groups had significantly elevated hepatic glycogen synthesis and glucokinase activity compared with the control group. Plasma total cholesterol and low density lipoprotein (LDL) cholesterol concentrations were significantly decreased by EAE and FA administration. These findings suggest that EAE and FA may be beneficial for treatment of type 2 diabetes because they regulate blood glucose levels by elevating glucokinase activity and production of glycogen in the liver.

  9. Cheek cell fatty acids reflect n-3 PUFA in blood fractions during linseed oil supplementation: a controlled human intervention study

    PubMed Central

    2013-01-01

    Background Adequate biomarkers for the dietary supply of fatty acids (FA) are FA of adipose tissue and blood fractions. In human studies, invasive sample collection is unpleasant for subjects. In contrast, cheek cell sampling can be considered as a non-invasive alternative to investigate FA status. The aim of this study was to analyze whether cheek cell FA composition reflect the supplementation of alpha-linolenic acid (ALA) using a linseed oil mixture compared to olive oil supplementation. Additionally, it was investigated if cheek cell FA composition correlates with the FA composition of plasma, red blood cells (RBC) and peripheral blood mononuclear cells (PBMC) before and during both interventions. Methods During a 10-week randomized, controlled, double-blind human intervention study, 38 subjects provided cheek cell and blood samples. After a two-week run-in period, the test group (n = 23) received 17 g/d of an ALA-rich linseed oil mixture, while the control group (n = 15) received 17 g/d of an omega-3 (n-3) polyunsaturated FA (PUFA)-free olive oil. Cheek cells and blood were collected on days 0, 7 and 56 of the 8-week intervention period. Results Compared to olive oil, the linseed oil intervention increased ALA and also the endogenously converted long-chain n-3 metabolites eicosatetraenoic-, eicosapentaenoic- and docosapentaenoic acid in cheek cells (P ≤ 0.05). Docosahexaenoic acid remained unchanged. Reflecting the treatment, the n-6/n-3 ratio decreased in the test group. In general, cheek cell FA reflected the changes of FA in blood fractions. Independent of treatment, significant correlations (P ≤ 0.05) of n-6 PUFA and n-3 PUFA between cheek cells and plasma, RBC and PBMC were found, except for linoleic acid and ALA. Conclusions The changes in FA composition of cheek cells confirmed that ALA from linseed oil increased endogenously derived n-3 PUFA in cheek cell lipids. These changes in cheek cells and their correlation to the respective

  10. Carbon isotopic fractionation in the biosynthesis of bacterial fatty acids. Ozonolysis of unsaturated fatty acids as a means of determining the intramolecular distribution of carbon isotopes

    NASA Astrophysics Data System (ADS)

    Monson, K. David; Hayes, J. M.

    1982-02-01

    Methods for the determination of 13C abundances at individual olefinic carbon positions have been developed, tested, and shown to perform accurately. (1) The double bond is oxidized with ozone; (2) silver oxide is used to cleave the resulting ozonide quantitatively to carboxylic-acid fragments; (3) a modified Schmidt decarboxylation is used to produce CO 2 quantitatively from the carboxyl groups of the separated cleavage products; (4) the CO 2 is utilized for mass spectrometric analysis. The results of intramolecular isotopic analyses are combined with molecular-average isotopic compositions determined by total combustion in order to show that fatty acids biosynthesized by Escherichia coli grown aerobically with glucose as the sole carbon source and harvested at late log phase are depleted by approximately 3%. in 13C relative to the glucose. This fractionation arises in the formation of acetylcoenzyme A by pyruvate dehydrogenase and is localized at the carboxyl position in the acetyl-CoA product. The isotopic order in that two-carbon subunit is carried through the biosynthesis of fatty acids so that alternate positions in the fatty-acid chains are depleted in 13C by an amount equal to twice the molecular-average depletion. The kinetic isotope effect at C-2 for pyruvate dehydrogenase in vivo is shown to be approximately 2.3%. While it appears that no other fractionation mechanism has controlled the overall depletion of 13C in these fatty acids, a separate process responsible for control of isotopic abundances in the carboxyl groups has been identified and described elsewhere [Monson K.D. and Hayes J.M. (1980) J. Biol. Chem. 255, 11435-11441]. It is concluded that kinetic, rather than thermodynamic, factors have controlled isotopic distributions in these cells and that kinetic factors will be dominant in most biological reactions.

  11. Capillary zone electrophoresis of soil humic acid fractions obtained by coupling size-exclusion chromatography and polyacrylamide gel electrophoresis.

    PubMed

    Cavani, Luciano; Ciavatta, Claudio; Trubetskaya, Olga E; Reznikova, Olga I; Afanas'eva, Gaida V; Trubetskoj, Oleg A

    2003-01-03

    Capillary zone electrophoresis (CZE) was used for characterisation of soil humic acid (HA) fractions obtained by coupling size-exclusion chromatography with polyacrylamide gel electrophoresis, on the basis of their molecular size and electrophoretic mobility. CZE was conducted using several low alkaline buffers as background electrolyte (BGE): 50 mM carbonate, pH 9.0; 50 mM phosphate, pH 8.5; 50 mM borate, pH 8.3; 50 mM Tris-borate+1 mM EDTA+7 M urea+0.1% sodium dodecyl sulphate (SDS), pH 8.3. Independently of BGE conditions, the effective electrophoretic mobility of HA fractions were in good agreement with their molecular size. The better resolution of HA were obtained in Tris-borate-EDTA buffer with urea and SDS. This results indicated that CZE, mostly with BGE-contained disaggregating agents, is useful for separating HAs in fractions with different molecular sizes.

  12. Some immunogenic acid biochemical properties of tumor-associated transplantation antigens (TATA) obtained in soluble form or solubilized from two methylcholanthrene-induced sarcomas, Meth A and CI-4.

    PubMed

    Rogers, M J; Law, L W

    1981-06-15

    The subcellular distribution of the tumor-associated transplantation antigens (TATAs) of two highly immunogenic methylcholanthrene-induced sarcomas, Meth A and CI-4, were compared. Most of the TATA of CI-4 was found in the soluble fraction (cytosol) of the cell while the TATA from Meth A was variably distributed between the membrane and cytosol. The soluble fraction TATAs from both tumors were very immunogenic and their strong immunity could not be influenced by administering antigen in a variety of protocols that altered the immune response in other systems. The soluble fraction and membrane-associated TATAs from both tumors could be specifically bound to liposomes in reconstitution experiments and the antigenicity of all of the TATAs was significantly enhanced when they were incorporated into these artificial membranes.

  13. Influence of protein formulation and carrier solution on asymmetrical flow field-flow fractionation: a case study of the plant-produced recombinant anthrax protective antigen pp-PA83.

    PubMed

    Palais, Caroline; Chichester, Jessica A; Manceva, Slobodanka; Yusibov, Vidadi; Arvinte, Tudor

    2015-02-01

    Asymmetrical flow field-flow fractionation (afFFF) was used to investigate the properties of a plant-produced anthrax toxin protective antigen, pp-PA83. The afFFF fractogram consisted of two main peaks with molar masses similar to the molecular mass of pp-PA83 monomer. afFFF carrier solutions strongly influenced the ratio and the intensity of the two main peaks. These differences indicate that conformation changes in the pp-PA83 molecule occurred during the afFFF analysis. Similar fractograms were obtained for different pp-PA83 formulations when the afFFF carrier solution and the protein formulation were the same (or very similar). The data show that in specific cases, afFFF could be used to study protein conformation and document the importance of studying the influence of the carrier solution on afFFF.

  14. Amino acid sequences of peptides from a tryptic digest of a urea-soluble protein fraction (U.S.3) from oxidized wool

    PubMed Central

    Corfield, M. C.; Fletcher, J. C.; Robson, A.

    1967-01-01

    1. A tryptic digest of the protein fraction U.S.3 from oxidized wool has been separated into 32 peptide fractions by cation-exchange resin chromatography. 2. Most of these fractions have been resolved into their component peptides by a combination of the techniques of cation-exchange resin chromatography, paper chromatography and paper electrophoresis. 3. The amino acid compositions of 58 of the peptides in the digest present in the largest amounts have been determined. 4. The amino acid sequences of 38 of these have been completely elucidated and those of six others partially derived. 5. These findings indicate that the parent protein in wool from which the protein fraction U.S.3 is derived has a minimum molecular weight of 74000. 6. The structures of wool proteins are discussed in the light of the peptide sequences determined, and, in particular, of those sequences in fraction U.S.3 that could not be elucidated. PMID:16742497

  15. Synergistic antioxidative effects of alkamides, caffeic acid derivatives, and polysaccharide fractions from Echinacea purpurea on in vitro oxidation of human low-density lipoproteins.

    PubMed

    Dalby-Brown, Lea; Barsett, Hilde; Landbo, Anne-Katrine R; Meyer, Anne S; Mølgaard, Per

    2005-11-30

    Preparations of Echinacea are widely used as alternative remedies to prevent the common cold and infections in the upper respiratory tract. After extraction, fractionation, and isolation, the antioxidant activity of three extracts, one alkamide fraction, four polysaccharide-containing fractions, and three caffeic acid derivatives from Echinacea purpurea root was evaluated by measuring their inhibition of in vitro Cu(II)-catalyzed oxidation of human low-density lipoprotein (LDL). The antioxidant activities of the isolated caffeic acid derivatives were compared to those of echinacoside, caffeic acid, and rosmarinic acid for reference. The order of antioxidant activity of the tested substances was cichoric acid > echinacoside > or = derivative II > or = caffeic acid > or = rosmarinic acid > derivative I. Among the extracts the 80% aqueous ethanolic extract exhibited a 10 times longer lag phase prolongation (LPP) than the 50% ethanolic extract, which in turn exhibited a longer LPP than the water extract. Following ion-exchange chromatography of the water extract, the majority of its antioxidant activity was found in the latest eluted fraction (H2O-acidic 3). The antioxidant activity of the tested Echinacea extracts, fractions, and isolated compounds was dose dependent. Synergistic antioxidant effects of Echinacea constituents were found when cichoric acid (major caffeic acid derivative in E. purpurea) or echinacoside (major caffeic acid derivative in Echinacea pallida and Echinacea angustifolia) were combined with a natural mixture of alkamides and/or a water extract containing the high molecular weight compounds. This contributes to the hypothesis that the physiologically beneficial effects of Echinacea are exerted by the multitude of constituents present in the preparations.

  16. Effect of Processing on phenolic acid composition of dough and bread fractions made from refined and whole-wheat flour of three wheat varieties

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In this study, we investigated the effect of bread-making on the assay of phenolic acids of whole and refined wheat from three wheat varieties, comparing refined (RF) and whole wheat (WW) flour, dough, and bread fractions. The efficacy of two common base hydrolysis methods for phenolic acid analysis...

  17. Sweet characterisation of prostate specific antigen using electrochemical lectin-based immunosensor assay and MALDI TOF/TOF analysis: Focus on sialic acid.

    PubMed

    Pihikova, Dominika; Pakanova, Zuzana; Nemcovic, Marek; Barath, Peter; Belicky, Stefan; Bertok, Tomas; Kasak, Peter; Mucha, Jan; Tkac, Jan

    2016-12-01

    The construction of a sensitive electrochemical lectin-based immunosensor for detection of a prostate specific antigen (PSA) is shown here. Three lectins with different carbohydrate specificities were used in this study to glycoprofile PSA, which is the most common biomarker for prostate cancer (PCa) diagnosis. The biosensor showed presence of α-L-fucose and α-(2,6)-linked terminal sialic acid within PSA´s glycan with high abundance, while only traces of α-(2,3)-linked terminal sialic acid were found. MALDI TOF/TOF mass spectrometry was applied to validate results obtained by the biosensor with a focus on determination of a type of sialic acid linkage by two methods. The first direct comparison of electrochemical immunosensor assay employing lectins for PSA glycoprofiling with mass spectrometric techniques is provided here and both methods show significant agreement. Thus, electrochemical lectin-based immunosensor has potential to be applied for prostate cancer diagnosis.

  18. Complementary use of flow and sedimentation field-flow fractionation techniques for size characterizing biodegradable poly(lactic acid) nanospheres

    PubMed Central

    Contado, Catia; Dalpiaz, Alessandro; Leo, Eliana; Zborowski, Maciej; Williams, P. Stephen

    2009-01-01

    Poly(lactic acid) nanoparticles were synthesized using a modified evaporation method, testing two different surfactants (sodium cholate and Pluronic F68) for the process. During their formulation the prodrug 5′-octanoyl-CPA (Oct-CPA) of the antiischemic N6-cyclopentyladenosine (CPA) was encapsulated. Three different purification methods were compared with respect to the influence of surfactant on the size characteristics of the final nanoparticle product. Flow and sedimentation field-flow fractionation techniques (FlFFF and SdFFF, respectively) were used to size characterize the five poly(lactic acid) particle samples. Two different combinations of carrier solution (mobile phase) were employed in the FlFFF analyses, while a solution of poly(vinyl alcohol) was used as mobile phase for the SdFFF runs. The separation performances of the two techniques were compared and the particle size distributions, derived from the fractograms, were interpreted with the support of observations by scanning electron microscopy. Some critical aspects, such as the carrier choice and the channel thickness determination for the FlFFF, have been investigated. This is the first comprehensive comparison of the two FFF techniques for characterizing non standard particulate materials. The two FFF techniques proved to be complementary and gave good, congruent and very useful information on the size distributions of the five poly(lactic acid) particle samples. PMID:17482199

  19. Chemical characterization of the inorganic fraction of aerosols and mechanisms of the neutralization of atmospheric acidity in Athens, Greece

    NASA Astrophysics Data System (ADS)

    Karageorgos, E. T.; Rapsomanikis, S.

    2007-06-01

    with inter-ionic correlations suggested that atmospheric ammonia is the major neutralizing agent of sulfate, while being insufficient to neutralize it to full extend. The formation of NH4NO3 is therefore not favored and additional contribution to the neutralization of acidity has been shown to be provided by Ca2+ and Mg2+. In the coarse particle fraction, the predominantly abundant Ca2+ has been found to correlate well with NO3- and SO42-, indicating its role as important neutralizing agent in this particle size range. The proximity of the location under study to the sea explains the important concentrations of salts with marine origin like NaCl and MgCl2 that were found in the coarse fraction, while chloride depletion in the gaseous phase was found to be limited to the fine particulate fraction. Total analyzed inorganic mass (elemental+ionic) was found to be ranging between approximately 25-33% of the total coarse particle mass and 35-42% of the total fine particle mass.

  20. Antigen-specific T cell-mediated suppression. V. H-2-linked genetic control of distinct antigen-specific defects in the production and activity of L-glutamic acid50-L-tyrosine50 suppressor factor

    PubMed Central

    1980-01-01

    The occurrence of distinct genetic defects affecting the generation of T cell-derived suppressor factor (TsF) or the suppressive activity of such TsF was investigated. For the synthetic polypeptide L-glutamic acid50-L-tyrosine50 (GT), it could be shown that the nonsuppressor strain A/J fails to produce suppressor T cells (Ts1) capable of GT-TsF generation upon challenge with GT. Conversely, B6, another nonsuppressor strain, produces GT-TsF active on other allogeneic strains such as A/J, but itself fails to be suppressed by this material. (B6A)F1 mice both make GT-TsF, and are suppressed by it. Further experiments revealed that the production of GT-TsF and the ability to be suppressed by GT-TsF are under the control of H-2-linked genes. Finally, the defect in GT-TsF activity in B6 mice was shown to be exquisitely antigen specific, in that this strain can be suppressed by a closely related TsF specific for L-glutamic acid60-L-alanine30-L- tyrosine10. It is suggested that H-2 (I) control of suppressor T cell (Ts) activity may reflect the involvement of I-A and I-C gene products in antigen presentation to Ts in analog with other T cell subsets, and that TsF function might also involve such presentation, in this case of the idiotypic structures of the TsF-combining site. Predictions deriving from this hypothesis are discussed, including the possibility that H-2 linked immune response genes regulate auto-anti-idiotypic responses in immune networks. PMID:6445400

  1. Boric acid, “carbonic” acid, and N-containing oxyacids in aqueous solution: Ab initio studies of structure, pKa, NMR shifts, and isotopic fractionations

    NASA Astrophysics Data System (ADS)

    Tossell, J. A.

    2005-12-01

    B(OH) 3 and CO 2 are acidic species of considerable geochemical importance, yet the microscopic nature of the acid dissociation reactions for these B and C species is not well understood. Quantum mechanical methods have recently been applied to the direct ab initio calculation of p Ka values for many organic and inorganic weak acids, but the B and C acids have not yet been considered in detail. In the present study, p Ka values are calculated quantum mechanically for the oxyacids B(OH) 3, H 2CO 3 and HNO 3, which have experimental first p Ka values of 9.2, 6.4 and -1.3, respectively. We calculate the gas-phase reaction free energies at the highly accurate CBS-QB3 ab initio quantum mechanical level and reaction free energies of hydration using a polarizable continuum method. Using a thermodynamic cycle corresponding to the simple dissociation process HA A - + H +, in aqueous solution, we calculate p Ka values of 21.6, 3.8 to 2.2 and -0.8 for the three oxyacids mentioned above, closely matching experiment only for HNO 3. The discrepancies with experiment arise from the more complex nature of the acid dissociation process for B(OH) 3, which involves the addition of H 2O to B(OH) 3 and formation of the B(OH) 4- anion, and from the instability of hypothetical H 2CO 3 compared to the proper hydrated reactant complex CO 2. . . H 2O. When the proper microscopic description of the reactants and products is used the calculated p Ka values for the three acids become 11.1, 7.2 and -0.8, in considerably better agreement with experiment for B(OH) 3 and CO 2. . . H 2O. Thus p Ka calculations using this approach are accurate enough to give information on the actual acid species present in solution and the details of their acid dissociation processes at the microscopic level. 11B and 13C-NMR chemical shifts are also calculated for the various species and compared to experiment. By comparison of our calculations with experiment it is apparent that the 13C-NMR chemical shift has

  2. Multiple amino acids in the capsid structure of canine parvovirus coordinately determine the canine host range and specific antigenic and hemagglutination properties.

    PubMed Central

    Chang, S F; Sgro, J Y; Parrish, C R

    1992-01-01

    Canine parvovirus (CPV) and feline panleukopenia virus (FPV) are over 98% similar in DNA sequence but have specific host range, antigenic, and hemagglutination (HA) properties which were located within the capsid protein gene. In vitro mutagenesis and recombination were used to prepare 16 different recombinant genomic clones, and viruses derived from those clones were analyzed for their in vitro host range, antigenic, and HA properties. The region of CPV from 59 to 91 map units determined the ability to replicate in canine cells. A complex series of interactions was observed among the individual sequence differences between 59 and 73 map units. The canine host range required that VP2 amino acids (aa) 93 and 323 both be the CPV sequence, and those two CPV sequences introduced alone into FPV greatly increased viral replication in canine cells. Changing any one of aa 93, 103, or 323 of CPV to the FPV sequence either greatly decreased replication in canine cells or resulted in an inviable plasmid. The Asn-Lys difference of aa 93 alone was responsible for the CPV-specific epitope recognized by monoclonal antibodies. An FPV-specific epitope was affected by aa 323. Amino acids 323 and 375 together determined the pH dependence of HA. Amino acids involved in the various specific properties were all around the threefold spikes of the viral particle. Images PMID:1331498

  3. Can the development and autolysis of lactic acid bacteria influence the cheese volatile fraction? The case of Grana Padano.

    PubMed

    Lazzi, Camilla; Povolo, Milena; Locci, Francesco; Bernini, Valentina; Neviani, Erasmo; Gatti, Monica

    2016-09-16

    In this study, the relationship between the dynamics of the growth and lysis of lactic acid bacteria in Grana Padano cheese and the formation of the volatile flavor compounds during cheese ripening was investigated. The microbial dynamics of Grana Padano cheeses that were produced in two different dairies were followed during ripening. The total and cultivable lactic microflora, community composition as determined by length heterogeneity-PCR (LH-PCR), and extent of bacterial lysis using an intracellular enzymatic activity assay were compared among cheeses after 2, 6 and 13months of ripening in two dairies. The evolution of whole and lysed microbiota was different between the two dairies. In dairy 2, the number of total cells was higher than that in dairy 1 in all samples, and the number of cells that lysed during ripening was lower. In addition, at the beginning of ripening (2months), the community structure of the cheese from dairy 2 was more complex and was composed of starter lactic acid bacteria (Lactobacillus helveticus and Lactobacillus delbrueckii) and NSLAB, possibly arising from raw milk, including Lactobacillus rhamnosus/Lactobacillus casei and Pediococcus acidilactici. On the other hand, the cheese from dairy 1 that ripened for 2months was mainly composed of the SLAB L. helveticus and L. delbrueckii. An evaluation of the free-DNA fraction through LH-PCR identified those species that had a high degree of lysis. Data on the dynamics of bacterial growth and lysis were evaluated with respect to the volatile profile and the organic acid content of the two cheeses after 13months of ripening, producing very different results. Cheese from dairy 1 showed a higher content of free fatty acids, particularly those deriving from milk fat lipolysis, benzaldehyde and organic acids, such as pGlu and citric. In contrast, cheese from dairy 2 had a greater amount of ketones, alcohols, hydrocarbons, acetic acid and propionic acid. Based on these results, we can conclude that

  4. [Effect of graded dietary protein rations on the amino acid content of crude protein in various parts of the gastrointestinal tract and blood fractions of laying hens].

    PubMed

    Gruhn, K; Wiefel, P

    1985-03-01

    Four groups of four colostomized laying hens each received rations only consisting of wheat, vitamins plus a mineral mixture and a graded daily feed supply of 110 g, 88 g, 66 g and 44 g. The determination of amino acids from faeces was carried out after hydrolysis from a 6-day sampling period from the 16 laying hybrids. At the end of the experiment the animals were slaughtered. The corpuscular fractions of the blood, the contents of crop and stomach as well as of the intestines were also hydrolysed and the amino acid content in the crude protein was determined. In addition, the content of free amino acids in the blood plasma was determined. The content of amino acids in the protein of the corpuscular blood fractions remained uninfluenced by the decreasing amino acid and energy supply. The content of free basic amino acids in the blood plasma decreased with the decreasing supply with amino acids and energy, whereas the content of free amino acids with branched chains and hydroxylized ones increased. The content of glutamic acid in the contents of crop and stomach changed considerably in comparison with feed protein. The amino acid values of the crude protein in the contents of the intestines and in faeces to a large extent differ considerably from those of the wheat fed and are approximate values of body protein. Deficient supply with amino acids and energy did not influence the apparent digestibility of the amino acids.

  5. [SPECTRAL AND ACID-BASE PROPERTIES OF HEMOLYMPH PLASMA AND ITS FRACTIONS FROM GASTROPOD PULMONATE MOLLUSC ACHATINA FULICA].

    PubMed

    Petrova, T A; Lianguzov, A Yu; Malygina, N M

    2016-01-01

    The set of normal biochemical indicators of the hemolymph plasma of gastropod pulmonate mollusc Achatinafulica is described. Comparative analysis of the whole plasma and its subfractions enriched and depleted of oxygen-carrying protein hemocyanin was performed by spectrophotometry and spectrofluorimetry methods. Individual features of the absorption spectra were analyzed using fourth derivatives. The optimum method for estimating protein concentration was chosen. To characterize acid-base properties of plasma hemolymph and its sub-fractions we calculated buffer capacity, equivalence points and pK values of dominant buffer groups. It is shown that the major role in maintaining the buffer capacity of hemolymph belongs to the bicarbonate system. These results are compared with data for Helix pomatia available in literature. In the future the indicators studied in this work will be used to develop ecotoxicological criteria for the environmental assessment.

  6. Antioxidant and antiatherogenic properties of phenolic acid and flavonol fractions of fruits of 'Amari' and 'Hallawi' date (Phoenix dactylifera L.) varieties.

    PubMed

    Borochov-Neori, Hamutal; Judeinstein, Sylvie; Greenberg, Amnon; Volkova, Nina; Rosenblat, Mira; Aviram, Michael

    2015-04-01

    Date (Phoenix dactylifera L.) fruit phenolic-acid or flavonol fractions were examined in vitro for antioxidant and antiatherogenic properties. Two fractions of each subgroup were prepared from two date varieties, 'Amari' and 'Hallawi', by solid phase extraction on C18. The fractions were analyzed for phenolics composition by RP-HPLC and tested for ferric-reducing antioxidant power, free radical scavenging capacity, inhibition of Cu(2+)-induced LDL oxidation, and enhancement of HDL-mediated cholesterol efflux from macrophages. All four fractions exhibited variable capacities to reduce ferric ions, scavenge radicals, and inhibit LDL oxidation. Flavonol fractions were considerably better inhibitors of LDL oxidation compared to phenolic acid fractions, with IC50's of 9-31 nmol GAE mL(-1) compared to 85-116 nmol GAE mL(-1), respectively. Only the flavonol fractions stimulated cholesterol removal from macrophages. Within each subgroup, the levels of all the activities varied with fraction composition. The results demonstrated strong structure-activity relationships for date phenolics and identified date flavonols as potential antiatherogenic bioactives.

  7. Distribution patterns of phthalic acid esters in soil particle-size fractions determine biouptake in soil-cereal crop systems

    PubMed Central

    Tan, Wenbing; Zhang, Yuan; He, Xiaosong; Xi, Beidou; Gao, Rutai; Mao, Xuhui; Huang, Caihong; Zhang, Hui; Li, Dan; Liang, Qiong; Cui, Dongyu; Alshawabkeh, Akram N.

    2016-01-01

    The use of wastewater irrigation for food crops can lead to presence of bioavailable phthalic acid esters (PAEs) in soils, which increase the potential for human exposure and adverse carcinogenic and non-cancer health effects. This study presents the first investigation of the occurrence and distribution of PAEs in a maize-wheat double-cropping system in a wastewater-irrigated area in the North China Plain. PAE levels in maize and wheat were found to be mainly attributed to PAE stores in soil coarse (250–2000 μm) and fine sand (53–250 μm) fractions. Soil particle-size fractions with higher bioavailability (i.e., coarse and fine sands) showed greater influence on PAE congener bioconcentration factors compared to PAE molecular structures for both maize and wheat tissues. More PAEs were allocated to maize and wheat grains with increased soil PAE storages from wastewater irrigation. Additional findings showed that levels of both non-cancer and carcinogenic risk for PAE congeners in wheat were higher than those in maize, suggesting that wheat food security should be prioritized. In conclusion, increased soil PAE concentrations specifically in maize and wheat grains indicate that wastewater irrigation can pose a contamination threat to food resources. PMID:27555553

  8. Fractionation of metals and As in sediments from a biosphere reserve (Odiel salt marshes) affected by acidic mine drainage.

    PubMed

    Morillo, José; Usero, José; Rojas, Raquel

    2008-04-01

    The Odiel salt marshes (Marismas del Odiel) are an important nature area declared a Biosphere Reserve, but they are greatly affected by pollution from the Odiel River. Surface sediments from this area were analysed using the latest version of the BCR sequential extraction procedure to determine the fractionation of As, Cd, Cr, Cu, Fe, Mn, Ni, Pb and Zn among four geochemical phases (acid-soluble, reducible, oxidisable and residual). The total content of each of the metals and As was also determined. The results showed high concentrations of As, Cd, Cu, Pb and Zn, with maximums of 791 mg kg(-1) of As, 8.5 mg kg(-1) of Cd, 2,740 mg kg(-1) of Cu, 1,580 mg kg(-1) of Pb and 3,920 mg kg(-1) of Zn. The concentrations of Cr, Mn and Ni were low since there are no sources of pollution by them in the area. A comparison of the metal and As levels with the sediment quality guidelines showed that the pollution is sufficient to produce noxious effects in aquatic organisms in most of the Odiel salt marshes. Based on the chemical distribution of the elements, it was found that Cd and Zn were the most mobile (i.e., elements that can pass easily into the water under changing environmental conditions). However, Cr, Fe, Ni and As were present in the greatest percentages in the residual fraction, which implies that these elements are strongly linked to the sediments.

  9. Distribution patterns of phthalic acid esters in soil particle-size fractions determine biouptake in soil-cereal crop systems

    NASA Astrophysics Data System (ADS)

    Tan, Wenbing; Zhang, Yuan; He, Xiaosong; Xi, Beidou; Gao, Rutai; Mao, Xuhui; Huang, Caihong; Zhang, Hui; Li, Dan; Liang, Qiong; Cui, Dongyu; Alshawabkeh, Akram N.

    2016-08-01

    The use of wastewater irrigation for food crops can lead to presence of bioavailable phthalic acid esters (PAEs) in soils, which increase the potential for human exposure and adverse carcinogenic and non-cancer health effects. This study presents the first investigation of the occurrence and distribution of PAEs in a maize-wheat double-cropping system in a wastewater-irrigated area in the North China Plain. PAE levels in maize and wheat were found to be mainly attributed to PAE stores in soil coarse (250–2000 μm) and fine sand (53–250 μm) fractions. Soil particle-size fractions with higher bioavailability (i.e., coarse and fine sands) showed greater influence on PAE congener bioconcentration factors compared to PAE molecular structures for both maize and wheat tissues. More PAEs were allocated to maize and wheat grains with increased soil PAE storages from wastewater irrigation. Additional findings showed that levels of both non-cancer and carcinogenic risk for PAE congeners in wheat were higher than those in maize, suggesting that wheat food security should be prioritized. In conclusion, increased soil PAE concentrations specifically in maize and wheat grains indicate that wastewater irrigation can pose a contamination threat to food resources.

  10. Combined HPLC/HPSEC study of Suwannee River Fulvic Acid adsorptive fractionation on α-aluminum oxide.

    PubMed

    Kreller, David I; Schlautman, Mark A; McGunigale, Samantha L

    2013-01-15

    A novel liquid chromatographic (LC) method with repeated injections of Suwannee River Fulvic Acid (SRFA) was used to investigate its adsorptive fractionation by synthetic α-Al(2)O(3). Eluent (i.e., non-retained) SRFA for each injection was monitored by two ultraviolet (UV) absorbance detection channels (300 and 365 nm) and one fluorescence detection channel (λ(ex)=350 nm, λ(em)=450 nm). Preferential adsorption of SRFA constituents was revealed by the different responses of the three LC detection channels. Samples of non-retained SRFA from injections of three independent replicate experiments were collected and aggregated for subsequent analysis by steady state ultraviolet-visible (UV/vis) absorption spectrometry and size exclusion chromatography (SEC). The ratio of absorbance at 254 and 204 nm, a surrogate for specific UV absorbance at 254 nm, increased with increasing injection number for the non-retained SRFA, indicating the preferential adsorption of SRFA constituents containing aromatic moieties. SEC analysis confirmed the preferential adsorption of higher molecular weight (MW) SRFA constituents as the non-adsorbed SRFA fractions increased in MW across the series of injections. The SEC results also suggested that certain SRFA constituents in the ca. 2-5 kDa MW range adsorbed in early injections were displaced by higher MW species (ca. 5-10 kDa) in later injections.

  11. Structure of a new acidic O-antigen of Proteus vulgaris O22 containing O-acetylated 3-acetamido-3,6-dideoxy-D-glucose.

    PubMed

    Toukach, F V; Bartodziejska, B; Senchenkova, S N; Wykrota, M; Shashkov, A S; Rozalski, A; Knirel, Y A

    1999-05-31

    The acidic O-specific polysaccharide of Proteus vulgaris O22 was studied using 1H and 13C NMR spectroscopy, including 2D COSY, TOCSY, NOESY, and H-detected 1H, 13C heteronuclear multiple-quantum coherence (HMQC) experiments, and the following structure for the branched pentasaccharide repeating unit was established: [sequence: see text] where Qui3NAc is 3-acetamido-3,6-dideoxyglucose, O-acetylation of QuiNAc at position 4 is stoichiometric and at position 2 nonstoichiometric. Serological relationships of P. vulgaris O22 with some other Proteus strains were substantiated on the level of the O-antigen structures.

  12. Manganese accumulation in membrane fractions of primary astrocytes is associated with decreased γ-aminobutyric acid (GABA) uptake, and is exacerbated by oleic acid and palmitate.

    PubMed

    Fordahl, Steve C; Erikson, Keith M

    2014-05-01

    Manganese (Mn) exposure interferes with GABA uptake; however, the effects of Mn on GABA transport proteins (GATs) have not been identified. We sought to characterize how Mn impairs GAT function in primary rat astrocytes. Astrocytes exposed to Mn (500 μM) had significantly reduced (3)H-GABA uptake despite no change in membrane or cytosolic GAT3 protein levels. Co-treatment with 100 μM oleic or palmitic acids (both known to be elevated in Mn neurotoxicity), exacerbated the Mn-induced decline in (3)H-GABA uptake. Mn accumulation in the membrane fraction of astrocytes was enhanced with fatty acid administration, and was negatively correlated with (3)H-GABA uptake. Furthermore, control cells exposed to Mn only during the experimental uptake had significantly reduced (3)H-GABA uptake, and the addition of GABA (50 μM) blunted cytosolic Mn accumulation. These data indicate that reduced GAT function in astrocytes is influenced by Mn and fatty acids accumulating at or interacting with the plasma membrane.

  13. Distribution of heavy metals by the molecular-weight fractions of humic acids in the soils of long-term field experiments

    NASA Astrophysics Data System (ADS)

    Karpukhin, A. I.; Bushuev, N. N.

    2007-03-01

    The influence of different agricultural treatments on the contents of Cd, Zn, and Co in the organic matter and humic acids of soddy-podzolic and dark gray forest soils was studied in long-term field experiments. The use of gel chromatography proved the complicated molecular-weight composition of the humic acids in all the studied soils. The contents of heavy metals and their distribution by the molecular-weight fractions of humic acids were determined in the experimental soils and in the virgin podzolic soils. The complex nature of organomineral compounds of Cd and Zn with humic acids was shown by means of gel filtration. The thermodynamic stability of the organomineral complexes increased with the increasing weight of the initial molecular-weight fractions of humic acids.

  14. Measurement of IgG, IgA and IgE antibodies to Dermatophagoides pteronyssinus by antigen-binding assay, using a partially purified fraction of mite extract (F4P1).

    PubMed Central

    Chapman, M D; Platts-Mills, T A

    1978-01-01

    An extract of Dermatophagoides pteronyssinus culture has been fractionated by chromatography on Sephadex G-100 and Pevikon block electrophoresis to obtain a partially purified allergen (F4P1). This preparation has a molecular weight of between 15--25,000 Dalton, migrates slowly on electrophoresis, and is colourless in solution. The skin-test reactivity of F1P1 was comparable to that of crude D. pteronyssinus extract. F4P1 was radio-labelled with 125I and used in an antigen-binding radioimmunoassay to measure IgG, IgA and IgE antibody (ab) to D. pteronyssinus. IgG, ab was detected in serum from 32/34 (94%) mite-allergic persons, and from 10/31 (30%) nonallergic persons. IgA ab and IgE ab were found in sera from 22/34 (65%) and 37/34 (79%) allergic persons respectively. Neither IgA nor IgE ab could be detected in sera from non-allergic persons. An excellent correlation was found between radioallergo-sorbent technique (RAST), using crude D. pteronyssinus extract and IgE-binding activity (BA) for F4P1, (r=0.94, P less than 0.001). The antigen-binding assay for IgE BA was as sensitive as RAST, but less sensitive than PK testing. There was a very good quantitative correlation between IgG BA and IgE BA (r = 0.84, P less than 0.001). IgG BA was shown to rise in the serum of three patients treated with injections of D. pteronyssinus extract. PMID:750116

  15. Compositional changes in the hydrophobic acids fraction of drainage water from different land management practices.

    PubMed

    Byrne, Corinna M P; Hayes, Michael H B; Kumar, Rajeev; Novotny, Etelvino H; Lanigan, Gary; Richards, Karl G; Fay, Deirdre; Simpson, Andre J

    2010-08-01

    Dissolved organic matter (DOM) can play a key role in many environmental processes, including carbon cycling, nutrient transport and the fates of contaminants and of agrochemicals. Hydrophobic acids (Ho), the major components of the DOM, were recovered from the drainage waters from well-drained (WDS) and poorly-drained (PDS) Irish grassland soils in lysimeters, amended with N fertiliser (F) and with bovine urine (U) and were studied using 1D and 2D solution-state Nuclear Magnetic Resonance (NMR) spectroscopy. The Diffusion Edited (DE) (1)H NMR spectra indicated that the Ho consisted largely of larger molecules, or of molecules that formed rigid aggregates, and the 1D and the 2D (Heteronuclear Multiple Quantum Coherence - HMQC, the Total Correlation Spectroscopy - TOCSY, and the Nuclear Overhauser Effect - NOESY) spectra indicated that the samples were composed of lignin residues, carbohydrates, protein/peptides, and aliphatic components derived from plant waxes/cuticular materials and from microbial lipids. The F amendments increased the concentrations of Ho in the waters by 1.5 and 2.5 times those in the controls in the cases of WDS and PDS, respectively. The lignin-derived components were increased by 50% and 300% in the cases of the Ho from the WDS and PDS, respectively. Applications of F + U decreased the losses of Ho, (compared to the F amendments alone) and very significantly decreased those of the lignin-derived materials, indicating that enhanced microbial activity from U gave rise to enhanced metabolism of the Ho components, and especially of lignin. In contrast the less biodegradable aliphatic components containing cuticular materials increased as the result of applications of F + U. This study helps our understanding of how management practices influence the movement of C between terrestrial and aquatic environments.

  16. The amino acid sequence of protein SCMK-B2C from the high-sulphur fraction of wool keratin

    PubMed Central

    Elleman, T. C.

    1972-01-01

    1. The amino acid sequence of a protein from the reduced and carboxymethylated high-sulphur fraction of wool has been determined. 2. The sequence of this S-carboxymethylkerateine (SCMK-B2C) of 151 amino acid residues displays much internal homology and an unusual residue distribution. Thus a ten-residue sequence occurs four times near the N-terminus and five times near the C-terminus with few changes. These regions contain much of the molecule's half-cystine, whereas between them there is a region of 19 residues that are mainly small and devoid of cystine and proline. 3. Certain models of the wool fibre based on its mechanical and physical properties propose a matrix of small compact globular units linked together to form beaded chains. The unusual distribution of the component residues of protein SCMK-B2C suggests structures in the wool-fibre matrix compatible with certain features of the proposed models. PMID:4678578

  17. Antinociceptive activity of Paederosidic Acid Methyl Ester (PAME) from the n-butanol fraction of Paederia scandens in mice.

    PubMed

    Chen, Yu-Feng; Huang, Ying; Tang, Wei-Zhong; Qin, Lu-Ping; Zheng, Han-chen

    2009-08-01

    Antinociceptive activity of Paederosidic Acid Methyl Ester (PAME), a chemical compound isolated from the n-butanol fraction of Paederia scandens, was evaluated in mice using chemical and thermal models of nociception. PAME given by intraperitoneal injection at doses of 20, 40 and 60 mg/kg produced significant inhibitions on chemical nociception induced by intraperitoneal acetic acid, subplantar formalin or capsaicin injections and on thermal nociception in the tail-flick test and the hot plate test. In the pentobarbital sodium-induced sleep time test and the open-field test, PAME neither significantly enhanced the pentobarbital sodium-induced sleep time nor impaired the motor performance, indicating that the observed antinociceptive activity of PAME was unlikely due to sedation or motor abnormality. Core body temperature measurement showed that PAME did not affect temperature within a 2-h period. Moreover, PAME-induced antinociception in the hot plate test was insensitive to naloxone or nimodipine but significantly antagonized by L-NAME (N (G)-nitro-L-arginine methyl ester), methylene blue and glibenclamide. These results suggested that PAME-produced antinociception was possibly related to the pathway of NO-cGMP-ATP sensitive K(+) channels, which merited further studies regarding the precise site and mechanisms of action.

  18. Zoledronic acid induces dose-dependent increase of antigen-specific CD8 T-cell responses in combination with peptide/poly-IC vaccine.

    PubMed

    Park, Hye-Mi; Cho, Hyun-Il; Shin, Chang-Ae; Shon, Hyun-Jung; Kim, Tai-Gyu

    2016-03-04

    Zoledronic acid (ZA) is used for treating osteoporosis and for preventing skeletal fractures in cancer patients suffering from myeloma and prostate cancer. It is also reported to directly induce cancer cell apoptosis and indirectly modulate T-cell immune response as an antitumor agent. In this study, the effect of ZA following peptide/polyinosinic-polycytidylic acid (poly-IC) vaccination was investigated in a murine tumor model. The combination of ZA with peptide/poly-IC vaccine showed a synergistic effect on the induction of antigen-specific CD8 T-cell response. Three consecutive intravenous administrations of ZA was defined to induce the highest CD8 T-cell response. Further, total splenocyte counts and antigen-specific CD8 T-cell response gradually increased depending on the dose of ZA. In tumor-bearing mice, ZA showed a dose-dependent decrease of growth and prolonged survival. Treatment with ZA only decreased the number of CD11b(+)Gr1(+) myeloid cells in blood. Our results demonstrate that the use of ZA could improve antitumor immune responses induced by the peptide/poly-IC vaccine.

  19. A simian virus 40 large T-antigen segment containing amino acids 1 to 127 and expressed under the control of the rat elastase-1 promoter produces pancreatic acinar carcinomas in transgenic mice.

    PubMed Central

    Tevethia, M J; Bonneau, R H; Griffith, J W; Mylin, L

    1997-01-01

    The simian virus 40 large T antigen induces tumors in a wide variety of tissues in transgenic mice, the precise tissues depending on the tissue specificity of the upstream region controlling T-antigen expression. Expression of mutant T antigens that contain a subset of the protein's activities restricts the spectrum of tumors induced. Others showed previously that expression of a mutant large T antigen containing the N-terminal 121 amino acids (T1-121) under control of the lymphotropic papovavirus promoter resulted in slow-growing choroid plexus tumors, whereas full-length T antigen under the same promoter induced rapidly growing CPR tumors, T-cell lymphomas, and B-cell lymphomas. In those instances, the alteration in tumor induction or progression correlated with inability of the mutant large T antigen to bind the tumor suppressor p53. In the study reported here, we investigated the capacity of an N-terminal T antigen segment (T1-127) expressed in conjunction with small t antigen under control of the rat elastase-1 (E1) promoter to induce pancreatic tumors. The results show that pancreases of transgenic mice expressing T1-127 and small t antigen display acinar cell dysplasia at birth that progresses to neoplasia. The average age to death in these mice is within the range reported for transgenic mice expressing full-length T antigen under control of the E1 promoter. These results indicate that sequestering p53 by binding is not required for the development of rapidly growing acinar cell carcinomas. In addition, we provide evidence that small t antigen is unlikely to be required. Finally, we show that the p53 protein in acinar cell carcinomas is wild type in conformation. PMID:9343166

  20. Microwave-assisted extraction of bound phenolic acids in bran and flour fractions from sorghum and maize cultivars varying in hardness.

    PubMed

    Chiremba, Constance; Rooney, Lloyd W; Beta, Trust

    2012-05-09

    To release bound phenolic acids, a microwave-assisted extraction procedure was applied to bran and flour fractions obtained from eight sorghum and eight maize cultivars varying in hardness. The procedure was followed by HPLC analysis, and the identities of phenolic acids were confirmed by MS/MS spectra. The extraction of sorghum and maize bound phenolic acids was done for 90 s in 2 M NaOH to release ferulic acid and p-coumaric acid from bran and flour. Two diferulic acids, 8-O-4'- and 8-5'-benzofuran form, were identified and quantitated in sorghum bran, and only the former was found in maize bran. The contents of ferulic acid and diferulic acids in sorghum bran were 416-827 and 25-179 μg/g, respectively, compared to 2193-4779 and 271-819 μg/g in maize. Phenolic acid levels of sorghum were similar between hard and soft cultivars, whereas those of maize differed significantly (p < 0.05) except for ferulic acid in flour. Sorghum phenolic acids were not correlated with grain hardness as measured using a tangential abrasive decortication device. Maize ferulic acid (r = -0.601, p < 0.01), p-coumaric acid (r = -0.668, p < 0.01), and 8-O-4'-diferulic acid (r = -0.629, p < 0.01) were significantly correlated with hardness.

  1. Acylated Glycosidic Acid Methyl Esters Generated from the Convolvulin Fraction of Rhizoma Jalapae Braziliensis by Treatment with Indium(III) Chloride in Methanol.

    PubMed

    Ono, Masateru; Oda, Satoko; Yasuda, Shin; Mineno, Tomoko; Okawa, Masafumi; Kinjo, Junei; Miyashita, Hiroyuki; Yoshimitsu, Hitoshi; Nohara, Toshihiro; Miyahara, Kazumoto

    2017-01-01

    Four hexaglycosides of methyl 3S,12S-dihydroxyhexadecanoate (1-4) were provided after treatment of the crude convolvulin fraction from Rhizoma Jalapae Braziliensis (the root of Ipomoea operculata (GOMES) MART., Convolvulaceae) with indium(III) chloride in methanol. The structures of 1-4 were elucidated on the basis of spectroscopic and chemical methods. Their sugar moieties were partially acylated with organic acids including (3S,9R)-3,6:6,9-diepoxydecanoic (exogonic) acid, (E)-2-methylbut-2-enoic (tiglic) acid, and isovaleric acid.

  2. Identification of linoleic acid, a main component of the n-hexane fraction from Dryopteris crassirhizoma, as an anti-Streptococcus mutans biofilm agent.

    PubMed

    Jung, Ji-Eun; Pandit, Santosh; Jeon, Jae-Gyu

    2014-01-01

    Dryopteris crassirhizoma is a semi-evergreen plant. Previous studies have shown the potential of this plant as an agent for the control of cariogenic biofilms. In this study, the main antibacterial components of the plant were identified by correlating gas chromatography-mass spectrometry data with the antibacterial activity of chloroform and n-hexane fractions and then evaluating the activity of the most potent antibacterial component against Streptococcus mutans UA159 biofilms. The most potent antibacterial component was linoleic acid, a main component of the n-hexane fraction. Linoleic acid reduced viability in a dose dependent manner and reduced biofilm accumulation during initial and mature biofilm formation. Furthermore, when the biofilms were briefly treated with linoleic acid (10 min/treatment, a total of six times), the dry weight of the biofilms was significantly diminished. In addition, the anti-biofilm activity of the n-hexane fraction was similar to that of linoleic acid. These results suggest that the n-hexane fraction of D. crassirhizoma and linoleic acid may be useful for controlling cariogenic biofilms.

  3. Suppression of antigen-specific antibody responses in mice exposed to perfluorooctanoic acid: Role of PPARα and T- and B-cell targeting.

    PubMed

    DeWitt, Jamie C; Williams, Wanda C; Creech, N Jonathan; Luebke, Robert W

    2016-01-01

    T-cell-dependent antibody responses (TDAR) are suppressed in female C57BL/6N mice exposed to ≥3.75 mg/kg of perfluorooctanoic acid (PFOA) for 15 days. To determine if suppression of humoral immunity by PFOA is peroxisome proliferator activated receptor alpha (PPARα)-dependent and if suppression is associated with specific targeting of T- or B-cells, three separate experiments were conducted: (1) female PPARα constitutive knockout (PPARα KO; B6.129S4-Ppar(tm1Gonz)N12) and wild-type controls (WT; C57BL/6-Tac) exposed to 0, 7.5, or 30 mg PFOA/kg for 15 days were immunized on Day 11 with a T-cell-dependent antigen and sera then collected for measures of antigen-specific IgM titers (TDAR) 5 days later; (2) female C57BL/6N WT mice exposed to 0, 0.94, 1.88, 3.75, or 7.5 mg PFOA/kg for 15 days were immunized with a T-cell-independent antigen on Day 11 and sera were then collected for analyses of antigen-specific IgM titers (TIAR) 7 days later; and (3) splenic lymphocyte phenotypes were assessed in unimmunized female C57BL/6N WT mice exposed to 0, 3.75, or 7.5 mg PFOA/kg for 10 days to investigate effects of PFOA in the absence of specific immunization. Separate groups of mice were immunized with a T-cell-dependent antigen after 11 days of exposure and splenic lymphocyte sub-populations were assessed after 13 or 15 days of exposure to assess numbers of stimulated cells. The results indicated that exposure to ≥1.88 mg PFOA/kg suppressed the TIAR; exposure to 30 mg PFOA/kg suppressed the TDAR in both PPARα KO and WT mice. The percentage of splenic B-cells was unchanged. Results obtained in the PPARα KO mice indicated that PPARα suppression of TDAR was independent of PPARα involvement. Suppression of the TIAR and the TDAR with minimal lymphocyte sub-population effects suggested that effects on humoral immunity are likely mediated by disruption of B-cell/plasma cell function.

  4. Minipool Caprylic Acid Fractionation of Plasma Using Disposable Equipment: A Practical Method to Enhance Immunoglobulin Supply in Developing Countries

    PubMed Central

    El-Ekiaby, Magdy; Vargas, Mariángela; Sayed, Makram; Gorgy, George; Goubran, Hadi; Radosevic, Mirjana; Burnouf, Thierry

    2015-01-01

    Background Immunoglobulin G (IgG) is an essential plasma-derived medicine that is lacking in developing countries. IgG shortages leave immunodeficient patients without treatment, exposing them to devastating recurrent infections from local pathogens. A simple and practical method for producing IgG from normal or convalescent plasma collected in developing countries is needed to provide better, faster access to IgG for patients in need. Methodology/Principal Findings IgG was purified from 10 consecutive minipools of 20 plasma donations collected in Egypt using single-use equipment. Plasma donations in their collection bags were subjected to 5%-pH5.5 caprylic acid treatment for 90 min at 31°C, and centrifuged to remove the precipitate. Supernatants were pooled, then dialyzed and concentrated using a commercial disposable hemodialyzer. The final preparation was filtered online by gravity, aseptically dispensed into storage transfusion bags, and frozen at <-20°C. The resulting preparation had a mean protein content of 60.5 g/L, 90.2% immunoglobulins, including 83.2% IgG, 12.4% IgA, and 4.4% IgM, and residual albumin. There was fourfold to sixfold enrichment of anti-hepatitis B and anti-rubella antibodies. Analyses of aggregates (<3%), prekallicrein (5-7 IU/mL), plasmin (26.3 mU/mL), thrombin (2.5 mU/mL), thrombin-like activity (0.011 U/g), thrombin generation capacity (< 223 nM), and Factor XI (<0.01 U/mL) activity, Factor XI/XIa antigen (2.4 ng/g) endotoxin (<0.5 EU/mL), and general safety test in rats showed the in vitro safety profile. Viral validation revealed >5 logs reduction of HIV, BVDV, and PRV infectivity in less than 15 min of caprylic acid treatment. Conclusions/Significance 90% pure, virally-inactivated immunoglobulins can be prepared from plasma minipools using simple disposable equipment and bag systems. This easy-to-implement process could be used to produce immunoglobulins from local plasma in developing countries to treat immunodeficient patients

  5. Evaluation of an integrated biorefinery based on fractionation of spent sulphite liquor for the production of an antioxidant-rich extract, lignosulphonates and succinic acid.

    PubMed

    Alexandri, Maria; Papapostolou, Harris; Komaitis, Michael; Stragier, Lutgart; Verstraete, Willy; Danezis, Georgios P; Georgiou, Constantinos A; Papanikolaou, Seraphim; Koutinas, Apostolis A

    2016-08-01

    Spent sulphite liquor (SSL) has been used for the production of lignosulphonates (LS), antioxidants and bio-based succinic acid. Solvent extraction of SSL with isopropanol led to the separation of approximately 80% of the total LS content, whereas the fermentations carried out using the pretreated SSL with isopropanol led to the production of around 19g/L of succinic acid by both Actinobacillus succinogenes and Basfia succiniciproducens. Fractionation of SSL via nanofiltration to separate the LS and solvent extraction using ethyl acetate to separate the phenolic compounds produced a detoxified sugar-rich stream that led to the production of 39g/L of succinic acid by B. succiniciproducens. This fractionation scheme resulted also in the production of 32.4g LS and 1.15g phenolic-rich extract per 100g of SSL. Both pretreatment schemes removed significant quantities of metals and heavy metals. This novel biorefinery concept could be integrated in acidic sulphite pulping mills.

  6. Distribution of rare earth elements in anionic, cationic and particulate fractions in boreal humus-rich streams affected by acid sulphate soils.

    PubMed

    Aström, Mats; Corin, Nina

    2003-01-01

    The abundance, fractionation and physicochemical forms of rare earth elements (REEs) were determined in five boreal humus-rich streams (dissolved organic carbon, DOC = 14-40 mg/l) affected by acid sulphate soils. The sampling was carried out during high-water flow in autumn when the acid sulphate soils are extensively flushed. The analytical procedures included ion-exchange experiments in field and ICP-MS determination. There was a general decrease in pH (range 4.5-6.2) and increase in the REE concentrations (La range 0.82-23 microg/l) as the proportion of the catchment cover of acid sulphate soils increased, explained by high amounts of REEs in the acidic runoff from such soils. In each stream, four different REE fractions were identified: (1) A cationic fraction, which is dominant in the REE-rich runoff from the acid sulphate soils and which is depleted in HREEs due to hydrochemical and/or geochemical processes, (2) an anionic fraction identified as humus-REE complexes, which in general is more abundant the higher the DOC concentrations and which also becomes increasingly abundant across the lanthanide series, (3) a fraction having a well-developed MREE enrichment, presumably consisting of colloidal REEs, and (4) a minor uncharacterised particle-associated fraction. The REE pool in the streams thus consists of several coexisting and contrasting REE species. The identification and quantification of such species is a prerequisite for the precise and accurate characterisation of the REE hydrochemistry of the streams.

  7. Antigen detection systems

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Infectious agents or their constituent parts (antigens or nucleic acids) can be detected in fresh, frozen, or fixed tissues or other specimens, using a variety of direct or indirect assays. The assays can be modified to yield the greatest sensitivity and specificity but in most cases a particular m...

  8. Antigen detection systems

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Infectious agents or their constituent parts (antigens or nucleic acids) can be detected in fresh, frozen, or fixed tissue using a variety of direct or indirect assays. The assays can be modified to yield the greatest sensitivity and specificity but in most cases a particular methodology is chosen ...

  9. [Isolation and physico-chemical characteristics of human cancerocerebral antigen].

    PubMed

    Prokopenko, P G; Borisenko, S A; Tatarinov, Iu S

    1984-01-01

    During gel filtration on Sephadex G-200 human cancerocerebral antigen (CCA) was eluted as two protein fractions with molecular mass of 135,000 and 270.000 daltons. Only one band of protein with molecular mass of about 15,000 daltons was noted after electrophoresis in 10% polyacrylamide gel containing SDS. As characteristic properties of CCA were recognized an electrophoretic polymorphism and a distinct trend to polymerization and isomeria. The antigen was not stained with dyes designed for staining base proteins, lipo-,glyco- and ferroproteins; CCA was thermostable (5 min at 80 degrees), it was inactivated by trypsin and protease but was resistant to pronase, hexokinase, alpha-amylase and beta-glucuronidase. A procedure was developed for isolation of CCA from brain, including fractionation with ammonium sulfate, ion exchange chromatography on DEAE-Sephadex A-50. The procedure enabled to obtain the CCA preparations suitable for radioimmunological, immunobiological assays and amino acid analyses.

  10. Expression of the surface antigen 4F2hc affects system-L-like neutral-amino-acid-transport activity in mammalian cells.

    PubMed Central

    Bröer, S; Bröer, A; Hamprecht, B

    1997-01-01

    Mammalian cells possess a variety of amino acid-transport systems with overlapping substrate specificity. System L is one of the major amino acid-transport systems of non-epithelial cells. By expression cloning we have recently demonstrated that the surface antigen 4F2hc (CD98) is a necessary component for expression of system-L-like amino acid-transport activity in C6-BU-1 rat glioma cells [Bröer, Bröer and Hamprecht (1995) Biochem. J. 312, 863-870]. 4F2hc mRNA was detected in CHO cells, COS cells, activated lymphocytes isolated from mouse spleen and primary cultures of astrocytes. In all these cell types, Na+-independent isoleucine transport was mediated by system L. No contribution of system y+L to isoleucine or arginine transport was detected in C6-BU-1 cells. In lymphocytes, both system-L-like amino acid-transport activity and 4F2hc mRNA levels increased after treatment with phorbol ester plus ionomycin. Antisense oligonucleotides caused modest inhibition of Na+-independent isoleucine transport in C6-BU-1 cells and primary cultures of astroglial cells, whereas arginine transport was unaffected. Overexpression of 4F2hc cDNA in CHO cells resulted in an increase in Na+-independent isoleucine transport. PMID:9182715

  11. BIOCHEMICAL STUDIES ON SO-CALLED SYPHILIS ANTIGEN.

    PubMed

    Noguchi, H; Bronfenbrenner, J

    1911-01-05

    of tissue is very variable. (c) Substances Soluble in Ether, Alcohol, and Aceton.-In this group are found varying amounts of fatty acids, both saturated and unsaturated, some neutral fats, cholesterin and many unidentified lipoidal bodies. This group causes either hemolysis or inhibition, of hemolysis. In other words, it is anticomplementary as well as hemolytic in the majority of preparations. At the same time, in some preparations it is, to a certain extent, antigenic. This great variation in the amounts of these substances in given extracts renders their presence in the antigen preparation undesirable. It is not denied, however, that, when added in adequate quantities, some of these substances may improve the activity of the antigenic lipoids. (d) Substances Insoluble in Aceton.-This group of substances consists of phosphatids. The best known among them is, of course, lecithin. Besides lecithin, however, there must be various other phosphatids present in this fraction. It will be noticed that the precipitate formed by mixing the ethereal solution with aceton contains a certain amount of lipoids insoluble in ether as well as in alcohol. Before the fractionation in aceton, all lipoids were soluble in ether or ethyl alcohol. Further analytical work on the nature of the phosphatids contained in this fraction is necessary. This fraction, in general, is more constant in amount in the various liver extracts. Biologically considered, it is the most important. It is usually non-hemolytic, frequently anticomplementary, but much more strongly antigenic than the other fractions. The antigenic strength varies with different preparations, being almost absent in the extracts derived from fatty livers. An aceton insoluble fraction may be strongly antigenic without any other auxiliary effects, or may be accompanied by an anticomplementary property. This fraction does not cause the so-called non-specific reaction with an active human serum. For these reasons it is recommended (as

  12. Serospecific antigens of Legionella pneumophila.

    PubMed Central

    Otten, S; Iyer, S; Johnson, W; Montgomery, R

    1986-01-01

    Serospecific antigens isolated by EDTA extraction from four serogroups of Legionella pneumophila were analyzed for their chemical composition, molecular heterogeneity by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and immunological properties. The antigens were shown to be lipopolysaccharides and to differ from the lipopolysaccharides of other gram-negative bacteria. The serospecific antigens contained rhamnose, mannose, glucosamine, and two unidentified sugars together with 2-keto-3-deoxyoctonate, phosphate, and fatty acids. The fatty acid composition was predominantly branched-chain acids with smaller amounts of 3-hydroxymyristic acid. The antigens contain periodate-sensitive groups; mannosyl residues were completely cleaved by periodate oxidation. Hydrolysis of the total lipopolysaccharide by acetic acid resulted in the separation of a lipid A-like material that cross-reacted with the antiserum to lipid A from Salmonella minnesota but did not comigrate with it on sodium dodecyl sulfate gels. None of the four antigens contained heptose. All of the antigen preparations showed endotoxicity when tested by the Limulus amebocyte lysate assay. The results of this study indicate that the serogroup-specific antigens of L. pneumophila are lipopolysaccharides containing an unusual lipid A and core structure and different from those of other gram-negative bacteria. Images PMID:3017918

  13. The Quinovic Acid Glycosides Purified Fraction from Uncaria tomentosa Protects against Hemorrhagic Cystitis Induced by Cyclophosphamide in Mice

    PubMed Central

    Dietrich, Fabrícia; Pietrobon Martins, Jerônimo; Kaiser, Samuel; Madeira Silva, Rodrigo Braccini; Rockenbach, Liliana; Albano Edelweiss, Maria Isabel; Ortega, George González; Morrone, Fernanda Bueno; Campos, Maria Martha; Battastini, Ana Maria Oliveira

    2015-01-01

    Uncaria tomentosa is widely used in folk medicine for the treatment of numerous diseases, such as urinary tract disease. Hemorrhagic cystitis (HE) is an inflammatory condition of the bladder associated with the use of anticancer drugs such as cyclophosphamide (CYP). Sodium 2-mercaptoethanesulfonate (Mesna) has been used to prevent the occurrence of HE, although this compound is not effective in established lesions. It has been demonstrated that the purinergic system is involved in several pathophysiological events. Among purinergic receptors, P2X7 deserves attention because it is involved in HE induced by CYP and, therefore, can be considered a therapeutic target. The objective of this study was to investigate the potential therapeutic effect of the quinovic acid glycosides purified fraction (QAPF) from U. tomentosa in the mouse model of CYP-induced HE. Pretreatment with QAPF not only had a protective effect on HE-induced urothelial damage (edema, hemorrhage and bladder wet weight) but was also able to control visceral pain, decrease IL-1β levels and down-regulates P2X7 receptors, most likely by inhibit the neutrophils migration to the bladder. This research clearly demonstrates the promising anti-inflammatory properties of QAPF, supporting its use as complementary therapy. QAPF represents a promising therapeutic option for this pathological condition. PMID:26154141

  14. Distribution of soil arsenic species, lead and arsenic bound to humic acid molar mass fractions in a contaminated apple orchard.

    PubMed

    Newton, Kimberly; Amarasiriwardena, Dulasiri; Xing, Baoshan

    2006-09-01

    Excessive application of lead arsenate pesticides in apple orchards during the early 1900s has led to the accumulation of lead and arsenic in these soils. Lead and arsenic bound to soil humic acids (HA) and soil arsenic species in a western Massachusetts apple orchard was investigated. The metal-humate binding profiles of Pb and As were analyzed with size exclusion chromatography-inductively coupled plasma mass spectrometry (SEC-ICP-MS). It was observed that both Pb and As bind "tightly" to soil HA molar mass fractions. The surface soils of the apple orchard contained a ratio of about 14:1 of water soluble As (V) to As (III), while mono-methyl (MMA) and di-methyl arsenic (DMA) were not detectable. The control soil contained comparatively very low levels of As (III) and As (V). The analysis of soil core samples demonstrated that As (III) and As (V) species are confined to the top 20 cm of the soil.

  15. The effect of hydrogen peroxide concentration and solid loading on the fractionation of biomass in formic acid.

    PubMed

    Dussan, K; Girisuta, B; Haverty, D; Leahy, J J; Hayes, M H B

    2014-10-13

    This study investigated the fractionation of biomass using a decomposing mixture of hydrogen peroxide-formic acid as a pretreatment for the biorefining of Miscanthus × giganteus and of sugarcane bagasse. The main parameters investigated were the hydrogen peroxide concentration (2.5, 5.0 and 7.5 wt%) and biomass loading (5.0 and 10.0 wt%). At the highest hydrogen peroxide concentration used (7.5 wt%), the energy released by the decomposition of the H2O2 could heat the reaction mixture up to 180 °C in a short time (6-16 min). As a result, highly delignified pulps, with lignin removal as high as 92 wt%, were obtained. This delignification process also solubilised a significant amount of pentosan (82-98 wt%) from the initial biomass feedstock, and the resulting pulp had a high cellulosic content (92 wt%). The biomass loading only affected the reaction rate of hydrogen peroxide decomposition. Various analytical methods, including Fourier transform infrared spectroscopy, and thermogravimetric and elemental analyses, characterized the lignin obtained.

  16. Saturated fatty acids induce post-transcriptional regulation of HAMP mRNA via AU-rich element-binding protein, human antigen R (HuR).

    PubMed

    Lu, Sizhao; Mott, Justin L; Harrison-Findik, Duygu Dee

    2015-10-02

    Iron is implicated in fatty liver disease pathogenesis. The human hepcidin gene, HAMP, is the master switch of iron metabolism. The aim of this study is to investigate the regulation of HAMP expression by fatty acids in HepG2 cells. For these studies, both saturated fatty acids (palmitic acid (PA) and stearic acid (SA)) and unsaturated fatty acid (oleic acid (OA)) were used. PA and, to a lesser extent, SA, but not OA, up-regulated HAMP mRNA levels, as determined by real-time PCR. To understand whether PA regulates HAMP mRNA at the transcriptional or post-transcriptional level, the transcription inhibitor actinomycin D was employed. PA-mediated induction of HAMP mRNA expression was not blocked by actinomycin D. Furthermore, PA activated HAMP 3'-UTR, but not promoter, activity, as shown by reporter assays. HAMP 3'-UTR harbors a single AU-rich element (ARE). Mutation of this ARE abolished the effect of PA, suggesting the involvement of ARE-binding proteins. The ARE-binding protein human antigen R (HuR) stabilizes mRNA through direct interaction with AREs on 3'-UTR. HuR is regulated by phosphorylation-mediated nucleo-cytoplasmic shuttling. PA activated this process. The binding of HuR to HAMP mRNA was also induced by PA in HepG2 cells. Silencing of HuR by siRNA abolished PA-mediated up-regulation of HAMP mRNA levels. PKC is known to phosphorylate HuR. Staurosporine, a broad-spectrum PKC inhibitor, inhibited both PA-mediated translocation of HuR and induction of HAMP expression. Similarly, rottlerin, a novel class PKC inhibitor, abrogated PA-mediated up-regulation of HAMP expression. In conclusion, lipids mediate post-transcriptional regulation of HAMP throughPKC- and HuR-dependent mechanisms.

  17. Sorption of Aldrich humic acid onto hematite: insights into fractionation phenomena by electrospray ionization with quadrupole time-of-flight mass spectrometry.

    PubMed

    Reiller, Pascal; Amekraz, Badia; Moulin, Christophe

    2006-04-01

    Sorption induced fractionation of purified Aldrich humic acid (PAHA) on hematite is studied through the modification of electrospray ionization (ESI) quadrupole time-of-flight (QToF) mass spectra of supernatants from retention experiments. The ESI mass spectra show an increase of the "mean molecular masses" of the molecules that constitutes humic aggregates. The low molecular weight fraction (LMWF; m/z < or = 600 Da) is preferentially sorbed compared to two other fractions. The resolution provided by ESI-QToF mass spectrometer in the low-mass range provided evidence of further fractionation induced by sorption within the LMWF. Among the two latter fractions, the high molecular weight fraction (HMWF; m/z approximately 1700 Da) seems to be more prone to sorption compared to the intermediate molecular weight fraction (IMWF; m/z approximately 900 Da). The IMWF seems to be more hydrophilic as it should be richer in O, N, and alkyl C from the proportion of even mass, and poorer in aromatic structures from mass defect analysis in ESI mass spectra.

  18. Point-Counterpoint: A Nucleic Acid Amplification Test for Streptococcus pyogenes Should Replace Antigen Detection and Culture for Detection of Bacterial Pharyngitis.

    PubMed

    Pritt, Bobbi S; Patel, Robin; Kirn, Thomas J; Thomson, Richard B

    2016-10-01

    Nucleic acid amplification tests (NAATs) have frequently been the standard diagnostic approach when specific infectious agents are sought in a clinic specimen. They can be applied for specific agents such as S. pyogenes, or commercial multiplex NAATs for detection of a variety of pathogens in gastrointestinal, bloodstream, and respiratory infections may be used. NAATs are both rapid and sensitive. For many years, S. pyogenes testing algorithms used a rapid and specific group A streptococcal antigen test to screen throat specimens, followed, in some clinical settings, by a throat culture for S. pyogenes to increase the sensitivity of its detection. Now S. pyogenes NAATs are being used with increasing frequency. Given their accuracy, rapidity, and ease of use, should they replace antigen detection and culture for the detection of bacterial pharyngitis? Bobbi Pritt and Robin Patel of the Mayo Clinic, where S. pyogenes NAATs have been used for well over a decade with great success, will explain the advantages of this approach, while Richard (Tom) Thomson and Tom Kirn of the NorthShore University HealthSystem will discuss their concerns about this approach to diagnosing bacterial pharyngitis.

  19. Novel prostate acid phosphatase-based peptide vaccination strategy induces antigen-specific T-cell responses and limits tumour growth in mice.

    PubMed

    Saif, Jaimy M S; Vadakekolathu, Jayakumar; Rane, Shraddha S; McDonald, Danielle; Ahmad, Murrium; Mathieu, Morgan; Pockley, A Graham; Durrant, Lindy; Metheringham, Rachael; Rees, Robert C; McArdle, Stephanie E B

    2014-04-01

    Treatment options for patients with advanced prostate cancer remain limited and rarely curative. Prostatic acid phosphatase (PAP) is a prostate-specific protein overexpressed in 95% of prostate tumours. An FDA-approved vaccine for the treatment of advanced prostate disease, PROVENGE® (sipuleucel-T), has been shown to prolong survival, however the precise sequence of the PAP protein responsible for the outcome is unknown. As the PAP antigen is one of the very few prostate-specific antigens for which there is a rodent equivalent with high homology, preclinical studies using PAP have the potential to be directly relevant to clinical setting. Here, we show three PAP epitopes naturally processed and presented in the context of HHDII/DR1 (114-128, 299-313, and 230-244). The PAP-114-128 epitope elicits CD4(+) and CD8(+) T-cell-specific responses in C57BL/6 mice. Furthermore, when immunised in a DNA vector format (ImmunoBody®), PAP-114-128 prevents and reduces the growth of transgenic adenocarcinoma of mouse prostate-C1 prostate cancer cell-derived tumours in both prophylactic and therapeutic settings. This anti-tumour effect is associated with infiltration of CD8(+) tumour-infiltrating lymphocytes and the generation of high avidity T cells secreting elevated levels of IFN-γ. PAP-114-128 therefore appears to be a highly relevant peptide on which to base vaccines for the treatment of prostate cancer.

  20. Separation of silver nanoparticles by hollow fiber flow field-flow fractionation: Addition of tannic acid into carrier liquid as a modifier.

    PubMed

    Saenmuangchin, Rattaporn; Mettakoonpitak, Jaruwan; Shiowatana, Juwadee; Siripinyanond, Atitaya

    2015-10-09

    A homemade hollow fiber flow-field fractionation (Hf-FlFFF) coupled with inductively coupled plasma mass spectrometry (ICP-MS) was set-up for silver nanoparticles (AgNPs) separation by using polysulfone hollow fiber membrane (30,000 MW cutoff) as a separation channel. Tannic acid and citrate stabilized AgNPs were synthesized and introduced into Hf-FlFFF. The effects of carrier liquid and stabilizing agent on retention behavior of AgNPs were investigated. Different elution behaviors were observed as follows: with 0.02% (w/v) FL-70, all of AgNPs were eluted from Hf-FlFFF but differences in retention behaviors were observed for AgNPs with tannic acid and citrate stabilizing agents; and with 30mM TRIS buffer, only tannic acid stabilized AgNPs were eluted from Hf-FlFFF, whereas citrate stabilized AgNPs were not eluted. In this work, tannic acid addition into carrier liquid was proposed to modify the surface of AgNPs and the surface of the membrane, and thereby adjusting the retention behaviors of AgNPs. Various concentrations of tannic acid were added into FL-70 and TRIS buffer. With the use of 0.1mM tannic acid in 30mM TRIS buffer as the carrier liquid, retention behaviors of both tannic acid stabilized- and citrate stabilized-AgNPs were similar and with similar fractionation recovery.

  1. A single amino acid substitution (R441A) in the receptor-binding domain of SARS coronavirus spike protein disrupts the antigenic structure and binding activity

    SciTech Connect

    He Yuxian . E-mail: yhe@nybloodcenter.org; Li Jingjing; Jiang Shibo

    2006-05-26

    The spike (S) protein of severe acute respiratory syndrome coronavirus (SARS-CoV) has two major functions: interacting with the receptor to mediate virus entry and inducing protective immunity. Coincidently, the receptor-binding domain (RBD, residues 318-510) of SAR-CoV S protein is a major antigenic site to induce neutralizing antibodies. Here, we used RBD-Fc, a fusion protein containing the RBD and human IgG1 Fc, as a model in the studies and found that a single amino acid substitution in the RBD (R441A) could abolish the immunogenicity of RBD to induce neutralizing antibodies in immunized mice and rabbits. With a panel of anti-RBD mAbs as probes, we observed that R441A substitution was able to disrupt the majority of neutralizing epitopes in the RBD, suggesting that this residue is critical for the antigenic structure responsible for inducing protective immune responses. We also demonstrated that the RBD-Fc bearing R441A mutation could not bind to soluble and cell-associated angiotensin-converting enzyme 2 (ACE2), the functional receptor for SARS-CoV and failed to block S protein-mediated pseudovirus entry, indicating that this point mutation also disrupted the receptor-binding motif (RBM) in the RBD. Taken together, these data provide direct evidence to show that a single amino acid residue at key position in the RBD can determine the major function of SARS-CoV S protein and imply for designing SARS vaccines and therapeutics.

  2. Evolution of the capsid protein genes of foot-and-mouth disease virus: antigenic variation without accumulation of amino acid substitutions over six decades.

    PubMed Central

    Martínez, M A; Dopazo, J; Hernández, J; Mateu, M G; Sobrino, F; Domingo, E; Knowles, N J

    1992-01-01

    The genetic diversification of foot-and-mouth disease virus (FMDV) of serotype C over a 6-decade period was studied by comparing nucleotide sequences of the capsid protein-coding regions of viruses isolated in Europe, South America, and The Philippines. Phylogenetic trees were derived for VP1 and P1 (VP1, VP2, VP3, and VP4) RNAs by using the least-squares method. Confidence intervals of the derived phylogeny (significance levels of nodes and standard deviations of branch lengths) were placed by application of the bootstrap resampling method. These procedures defined six highly significant major evolutionary lineages and a complex network of sublines for the isolates from South America. In contrast, European isolates are considerably more homogeneous, probably because of the vaccine origin of several of them. The phylogenetic analysis suggests that FMDV CGC Ger/26 (one of the earliest FMDV isolates available) belonged to an evolutionary line which is now apparently extinct. Attempts to date the origin (ancestor) of the FMDVs analyzed met with considerable uncertainty, mainly owing to the stasis noted in European viruses. Remarkably, the evolution of the capsid genes of FMDV was essentially associated with linear accumulation of silent mutations but continuous accumulation of amino acid substitutions was not observed. Thus, the antigenic variation attained by FMDV type C over 6 decades was due to fluctuations among limited combinations of amino acid residues without net accumulation of amino acid replacements over time. PMID:1316467

  3. Nucleic acid sequence of an internal image-bearing monoclonal anti-idiotype and its comparison to the sequence of the external antigen.

    PubMed Central

    Bruck, C; Co, M S; Slaoui, M; Gaulton, G N; Smith, T; Fields, B N; Mullins, J I; Greene, M I

    1986-01-01

    The monoclonal anti-idiotypic antibody (mAb2) 87.92.6 directed against the 9B.G5 antibody specific for the virus neutralizing epitope on the mammalian reovirus type 3 hemagglutinin was previously demonstrated to express an internal image of the receptor binding epitope of the reovirus type 3. Furthermore, this mAb2 has autoimmune reactivity to the cell surface receptor of the reovirus. The nucleotide and deduced amino acid sequences of the 87.92.6 mAb2 heavy and light chains are described in this report. The sequence analysis reveals that the same heavy chain variable and joining (VH and JH) gene segments are used by the 87.92.6 anti-idiotypic mAb2 and by the dominant idiotypes of the BALB/c anti-GAT (cGAT) and anti-NP (NPa) responses. [GAT; random polymer that is 60% glutamic acid, 30% alanine, and 10% tyrosine. NP; (4-hydroxy-3-nitrophenyl)-acetyl.] Despite extensive homology at the level of the heavy chain variable regions, the NPa positive BALB/c anti-NP monoclonal antibody 17.2.25 binds neither 9B.G5 nor the cellular receptor for the hemagglutinin. Amino acid sequence comparison between the viral hemagglutinin and the 87.92.6 mAb2 light chain "internal image," reveals an area of significant homology indicating that antigen mimicry by antibodies may be achieved by sharing primary structure. PMID:2428036

  4. Characterization of a 14,000 dalton antigen of Dirofilaria immitis infective third stage larvae

    SciTech Connect

    Fuller, S.A.; Cachia, P.J.; Wong, M.M.; Hurrell, J.G.R.

    1986-05-01

    Immunogenic proteins of Dirofilaria immitis (canine heartworm) were identified by probing extracts of adult worms or their excretory-secretory proteins (ESP) blotted to nitrocellulose following SDS-PAGE with control or infected dog sera. A 14,000 dalton antigen (a prominent component of ESP by protein staining) was consistently recognized both in extracts and ESP by dog sera as early as three months post infection. This indicates a larval origin for the antigen since no adult worms are present until approximately five months post infection. Monoclonal antibodies (MAbs) prepared against the 14,000 dalton antigen confirmed by immunoblotting that this antigen is expressed by infective third stage larvae, adults and microfilariae and is present intact in the sera of infected dogs. Surface-labelling of whole adult D. immitis with Na/sup 125/I produced radiolabelled antigens closely corresponding to those of ESP. An anti-14,000 dalton MAb was able to immunoprecipitate radiolabelled antigen which strongly suggest a surface or membrane location in the intact organism. Gel filtration data suggests that the protein is a native monomer. A MAb-affinity column has been used to purify the 14,000 dalton antigen to at least 98% homogeneity in one step from crude worm extracts. Further fractionation by HPLC yields a homogeneous preparation. Amino acid analysis and the N-terminal amino acid sequence data will be presented.

  5. Effects of soluble and insoluble fractions from bilberries, black currants, and raspberries on short-chain fatty acid formation, anthocyanin excretion, and cholesterol in rats.

    PubMed

    Jakobsdottir, Greta; Nilsson, Ulf; Blanco, Narda; Sterner, Olov; Nyman, Margareta

    2014-05-14

    Dietary fiber and flavonoids, important components in berries, are suggested to improve metabolic health. This study investigates whether soluble and insoluble fractions isolated from bilberry, black currant, and raspberry affect the formation of short-chain fatty acids (SCFAs), uptake and excretion of flavonoids, and levels of cholesterol differently. Cecal SCFA pools were higher in rats fed the soluble than the insoluble fractions (525 vs 166 μmol, P < 0.001), whereas higher concentrations of butyric acid were found in the distal colon and serum of rats fed the insoluble fractions (5 vs 3 μmol/g and 58 vs 29 μmol/L, respectively, P < 0.001). The soluble bilberry fraction gave lower amounts of liver cholesterol (56 mg) than the other berry fractions (87 ± 5 mg), formed the highest amount of SCFAs (746 vs 266 ± 21 μmol), and contributed the highest intake of anthocyanins. Cyanidin-3-glucoside monoglucuronide was detected in the urine of all groups, whereas anthocyanins were found only in groups fed soluble black currant and raspberry.

  6. Effects of Quality Composts and Other Organic Amendments and Their Humic and Fulvic Acid Fractions on the Germination and Early Growth of Slickspot Peppergrass (Lepidium Papilliferum) and Switchgrass in Various Experimental Conditions

    DTIC Science & Technology

    2008-09-01

    which appear slightly different from GCC-DOM, and the possible presence of benzoic acid derivatives such as phthalic and salicylic acids . 4...FULVIC ACID FRACTIONS ON THE GERMINATION AND EARLY GROWTH OF SLICKSPOT PEPPERGRASS (LEPIDIUM PAPILLIFERUM) AND SWITCHGRASS IN VARIOUS EXPERIMENTAL...Effects of Quality Composts and Other Organic Amendments and Their Humic and Fulvic Acid Fractions on the Germination and Early Growth of Slickspot

  7. Selective fermentation of carbohydrate and protein fractions of Scenedesmus, and biohydrogenation of its lipid fraction for enhanced recovery of saturated fatty acids.

    PubMed

    Lai, YenJung Sean; Parameswaran, Prathap; Li, Ang; Aguinaga, Alyssa; Rittmann, Bruce E

    2016-02-01

    Biofuels derived from microalgae have promise as carbon-neutral replacements for petroleum. However, difficulty extracting microalgae-derived lipids and the co-extraction of non-lipid components add major costs that detract from the benefits of microalgae-based biofuel. Selective fermentation could alleviate these problems by managing microbial degradation so that carbohydrates and proteins are hydrolyzed and fermented, but lipids remain intact. We evaluated selective fermentation of Scenedesmus biomass in batch experiments buffered at pH 5.5, 7, or 9. Carbohydrates were fermented up to 45% within the first 6 days, protein fermentation followed after about 20 days, and lipids (measured as fatty acid methyl esters, FAME) were conserved. Fermentation of the non-lipid components generated volatile fatty acids, with acetate, butyrate, and propionate being the dominant products. Selective fermentation of Scenedesmus biomass increased the amount of extractable FAME and the ratio of FAME to crude lipids. It also led to biohydrogenation of unsaturated FAME to more desirable saturated FAME (especially to C16:0 and C18:0), and the degree of saturation was inversely related to the accumulation of hydrogen gas after fermentation. Moreover, the microbial communities after selective fermentation were enriched in bacteria from families known to perform biohydrogenation, i.e., Porphyromonadaceae and Ruminococcaceae. Thus, this study provides proof-of-concept that selective fermentation can improve the quantity and quality of lipids that can be extracted from Scenedesmus.

  8. Characterization of fulvic acid fractions of dissolved organic matter during ice-out in a hyper-eutrophic, coastal pond in Antarctica

    NASA Astrophysics Data System (ADS)

    Cawley, Kaelin M.; McKnight, Diane M.; Miller, Penney; Cory, Rose; Fimmen, Ryan L.; Guerard, Jennifer; Dieser, Markus; Jaros, Christopher; Chin, Yu-Ping; Foreman, Christine

    2013-12-01

    Dissolved humic material (HDOM) is ubiquitous to all natural waters and its source material influences its chemical structure, reactivity, and bioavailability. While terrestrially derived HDOM reference materials distributed by the International Humic Substances Society (IHSS) have been readily available to engineering and scientific communities, a microbially derived reference HDOM was not, despite the well-characterized differences in the chemistry and reactivity of HDOM derived from terrestrial versus microbial sources. To address this gap, we collected a microbial reference fulvic acid from Pony Lake (PLFA) for distribution through the IHSS. Pony Lake is a saline coastal pond on Ross Island, Antarctica, where the landscape is devoid of terrestrial plants. Sample collection occurred over a 17-day period in the summer season at Pony Lake. During this time, the dissolved organic carbon (DOC) concentrations increased nearly two-fold, and the fulvic acid fraction (collected using the XAD-8 method) accounted for 14.6% of the DOC. During the re-concentration and desalting procedures we isolated two other chemically distinct fulvic acid fractions: (1) PLFA-2, which was high in carbohydrates and (2) PLFA-CER, which was high in nitrogen. The chemical characteristics (elemental analysis, optical characterization with UV-vis and fluorescence spectroscopy, and 13C NMR spectroscopy) of the three fulvic acid fractions helped to explain their behavior during isolation.

  9. Comparison of Xpert Flu rapid nucleic acid testing with rapid antigen testing for the diagnosis of influenza A and B.

    PubMed

    DiMaio, Michael A; Sahoo, Malaya K; Waggoner, Jesse; Pinsky, Benjamin A

    2012-12-01

    Influenza infections are associated with thousands of hospital admissions and deaths each year. Rapid detection of influenza is important for prompt initiation of antiviral therapy and appropriate patient triage. In this study the Cepheid Xpert Flu assay was compared with two rapid antigen tests, BinaxNOW Influenza A & B and BD Directigen EZ Flu A+B, as well as direct fluorescent antibody testing for the rapid detection of influenza A and B. Using real-time, hydrolysis probe-based, reverse transcriptase PCR as the reference method, influenza A sensitivity was 97.3% for Xpert Flu, 95.9% for direct fluorescent antibody testing, 62.2% for BinaxNOW, and 71.6% for BD Directigen. Influenza B sensitivity was 100% for Xpert Flu and direct fluorescent antibody testing, 54.5% for BinaxNOW, and 48.5% for BD Directigen. Specificity for influenza A was 100% for Xpert Flu, BinaxNOW, and BD Directigen, and 99.2% for direct fluorescent antibody testing. All methods demonstrated 100% specificity for influenza B. These findings support the use of the Xpert Flu assay in settings requiring urgent diagnosis of influenza A and B.

  10. Effects of specific amino acid changes on the antigenicity of hemagglutinin molecules of avian influenza isolates from Mexico

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Amino acid (aa) changes between the hemagglutinin (HA) proteins of a vaccine avian influenza virus and more recent field isolates were detected following prolonged vaccination of Mexican poultry. Using site-directed mutagenesis and reverse genetics (rg), viruses containing identical backbones but d...

  11. Effect of mineral surface properties (alumina, kaolinite) on the sorptive fractionation mechanisms of soil fulvic acids: Molecular-scale ESI-MS studies

    NASA Astrophysics Data System (ADS)

    Fleury, Guillaume; Del Nero, Mirella; Barillon, Rémi

    2017-01-01

    We addressed the effects of mineral surface properties (kaolinite versus Al-oxide) on the sorption-driven fractionation of a soil fulvic acid (FA) at acidic pH, mainly by means of ESI(-)-FTMS analysis of initial and supernatant solutions of FA sorption batch experiments. The MS data provided clear molecular-scale evidence of distinct mechanisms and molecular parameters controlling the FA fractionation upon its sorption on clay and oxide surfaces, respectively. Identification of sorbing and not-sorbing FA compounds in kaolinite-solution systems revealed a weak fractionation among members of sbnd CO2 series of aliphatics or not-condensed aromatics (NCAs) at pH 3.8, and almost no sorption of poorly-oxygenated polycyclic aromatic compounds (PACs) and NCAs. This first molecular-scale description of a FA fractionation in a clay-solution system suggests that H-bonding with low affinity sites (aluminol/silanol) on the basal planes of the clay particles is the main mechanism of sorption. Due to the predominance of such weak and poorly-selective mechanism, the sorption of aliphatic and NCA molecules bearing oxygenated functionalities was prevented at pH 5, due to dissolved Al competing successfully for their coordination. In contrast, a strong FA fractionation was observed onto alumina, with a preferential retention of PACs and highly-oxygenated aliphatics and NCAs. The major part of the poorly oxygenated aliphatics was left in solution. The sorption degree of NCAs and aliphatics was strongly correlated with molecular acidity. For PACs and poorly-oxygenated NCAs, the sorption was driven by reactions of surface ligand exchange (for the most oxygenated compounds) or by hydrophobic interactions (for the least oxygenated compounds).

  12. Sorption of selected organic compounds from water to a peat soil and its humic-acid and humin fractions: Potential sources of the sorption nonlinearity

    USGS Publications Warehouse

    Chiou, C.T.; Kile, D.E.; Rutherford, D.W.; Sheng, G.; Boyd, S.A.

    2000-01-01

    The sorption isotherms of ethylene dibromide (EDB), diuron (DUN), and 3,5-dichlorophenol (DCP) from water on the humic acid and humin fractions of a peat soil and on the humic-acid of a muck soil have been measured. The data were compared with those of the solutes with the whole peat from which the humic-acid (HA) and humin (HM) fractions were derived and on which the sorption of the solutes exhibited varying extents of nonlinear capacities at low relative concentrations (C(e)/S(w)). The HA fraction as prepared by the density-fractionated method is relatively pure and presumably free of high- surface-area carbonaceous material (HSACM) that is considered to be responsible for the observed nonlinear sorption for nonpolar solutes (e.g., EDB) on the peat; conversely, the base-insoluble HM fraction as prepared is presumed to be enriched with HSACM, as manifested by the greatly higher BET- (N2) surface area than that of the whole peat. The sorption of EDB on HA exhibits no visible nonlinear effect, whereas the sorption on HM shows an enhanced nonlinearity over that on the whole peat. The sorption of polar DUN and DCP on HA and HM display nonlinear effects comparable with those on the whole peat; the effects are much more significant than those with nonpolar EDB. These results conform to the hypothesis that adsorption onto a small amount of strongly adsorbing HSACM is largely responsible for the nonlinear sorption of nonpolar solutes on soils and that additional specific interactions with the active groups of soil organic matter are responsible for the generally higher nonlinear sorption of the polar solutes.

  13. Distribution of metals in acid-base-neutral fractions of Cerro Negro 550-700 degree C distillate and 700 degree C+ residue

    SciTech Connect

    Pearson, C.D.; Green, J.A.; Green, J.B.; Anderson, R.P. )

    1989-04-01

    Heavy crude oils generally have high metals content and are difficult to upgrade. Vanadium and nickel are among the most common metals and can cause catalyst poisoning and corrosion during processing. The continuing decrease in reserves of conventional crude oil has made it increasingly important that new techniques be developed for demetallization to permit upgrading and use of these difficult resources. Knowledge of the chemical composition of nickel and vanadium complexes in petroleum will assist the process designer to devise better demetallization processes. This paper describes a new approach to the speciation of metals in crude oils and residues. The Cerro Negro 550-700{degree}C distillate and 700{degree}C+ residue are broken down into acid, base, and neutral fractions and the nickel, vanadium, and iron determined in each fraction. An attempt is made to answer the questions: (1) is distribution governed by the chemical structure of the metalloorganic molecules; (2) within a given chemical fraction, are subfractions present which contain a major portion of the total metal content; (3) does the particular metal (i.e., Ni vs V) affect the separation behavior This paper is the sixth and final chapter in a book describing the development of analytical methodology for the analytical characterization of Cerro Negro Orinoco belt crude oil. Chapters already published address the distillation and determination of routine physical and chemical properties, the separation of acid, base, and neutral fractions and saturate/aromatic split of the neutral fractions, and the detailed separation and analysis of sulfur compounds and aromatic hydrocarbons. The chapters on detailed separation and analysis of acidic and basic compounds are currently being written. They are based on previously reported analytical procedures.

  14. Comparison of the phosphorus and mineral concentrations in bran and abraded kernel fractions of a normal barley (Hordeum vulgare) cultivar versus four low phytic acid isolines.

    PubMed

    Liu, Keshun; Peterson, Kevin L; Raboy, Victor

    2007-05-30

    Phytic acid consists of 65-80% of the total phosphorus (P) in cereal grains. Its salts are concentrated in the germ and aleurone layers, which are typically removed during milling. We hypothesize that concentrations of different types of P and minerals in milled products will be greatly altered in low phytic acid (lpa) barleys. Seeds of cv. Harrington (control) and four lpa isolines-lpa1-1, lpa2-1, lpa3-1, and M955-were abraded by a laboratory method into five surface layer and four remaining kernel fractions. Results show that phytic acid in the four lpa lines ranged from 75% to 5% of the control. The decrease in phytic acid P concentration was matched almost equally by an increase in inorganic P, so that the rest of P (the sum of all P-containing compounds other than phytic acid P and inorganic P) and total P levels remained relatively unchanged among the five genotypes. These trends were also observed for the processed fractions. The major mineral elements in barley seeds were P, K, Mg, S, and Ca, while minor ones were Fe, Zn, Mn, Cu, and Ba. All types of P and other minerals measured were generally concentrated in the outer layers of the grain. Although there were substantial differences in mineral contents of bran fractions among genotypes, the level of phytic acid P had little effect on mineral contents in whole or abraded kernels. One major exception was Fe, which had the highest level in all tissues of M955 genotype. The above findings were all confirmed by analyzing another set of barley samples grown in a different environment. Thus, in general, breeding lpa barleys does not lead to reduced mineral contents in whole grains or elevated mineral levels in milled products.

  15. Fractionation of Cu and Zn isotopes during adsorption onto amorphous Fe(III) oxyhydroxide: Experimental mixing of acid rock drainage and ambient river water

    USGS Publications Warehouse

    Balistrieri, L.S.; Borrok, D.M.; Wanty, R.B.; Ridley, W.I.

    2008-01-01

    Fractionation of Cu and Zn isotopes during adsorption onto amorphous ferric oxyhydroxide is examined in experimental mixtures of metal-rich acid rock drainage and relatively pure river water and during batch adsorption experiments using synthetic ferrihydrite. A diverse set of Cu- and Zn-bearing solutions was examined, including natural waters, complex synthetic acid rock drainage, and simple NaNO3 electrolyte. Metal adsorption data are combined with isotopic measurements of dissolved Cu (65Cu/63Cu) and Zn (66Zn/64Zn) in each of the experiments. Fractionation of Cu and Zn isotopes occurs during adsorption of the metal onto amorphous ferric oxyhydroxide. The adsorption data are modeled successfully using the diffuse double layer model in PHREEQC. The isotopic data are best described by a closed system, equilibrium exchange model. The fractionation factors (??soln-solid) are 0.99927 ?? 0.00008 for Cu and 0.99948 ?? 0.00004 for Zn or, alternately, the separation factors (??soln-solid) are -0.73 ?? 0.08??? for Cu and -0.52 ?? 0.04??? for Zn. These factors indicate that the heavier isotope preferentially adsorbs onto the oxyhydroxide surface, which is consistent with shorter metal-oxygen bonds and lower coordination number for the metal at the surface relative to the aqueous ion. Fractionation of Cu isotopes also is greater than that for Zn isotopes. Limited isotopic data for adsorption of Cu, Fe(II), and Zn onto amorphous ferric oxyhydroxide suggest that isotopic fractionation is related to the intrinsic equilibrium constants that define aqueous metal interactions with oxyhydroxide surface sites. Greater isotopic fractionation occurs with stronger metal binding by the oxyhydroxide with Cu > Zn > Fe(II).

  16. Fractionation of Cu and Zn isotopes during adsorption onto amorphous Fe(III) oxyhydroxide: Experimental mixing of acid rock drainage and ambient river water

    NASA Astrophysics Data System (ADS)

    Balistrieri, Laurie S.; Borrok, David M.; Wanty, Richard B.; Ridley, W. Ian

    2008-01-01

    Fractionation of Cu and Zn isotopes during adsorption onto amorphous ferric oxyhydroxide is examined in experimental mixtures of metal-rich acid rock drainage and relatively pure river water and during batch adsorption experiments using synthetic ferrihydrite. A diverse set of Cu- and Zn-bearing solutions was examined, including natural waters, complex synthetic acid rock drainage, and simple NaNO3 electrolyte. Metal adsorption data are combined with isotopic measurements of dissolved Cu (65Cu/63Cu) and Zn (66Zn/64Zn) in each of the experiments. Fractionation of Cu and Zn isotopes occurs during adsorption of the metal onto amorphous ferric oxyhydroxide. The adsorption data are modeled successfully using the diffuse double layer model in PHREEQC. The isotopic data are best described by a closed system, equilibrium exchange model. The fractionation factors (αsoln-solid) are 0.99927 ± 0.00008 for Cu and 0.99948 ± 0.00004 for Zn or, alternately, the separation factors (Δsoln-solid) are -0.73 ± 0.08‰ for Cu and -0.52 ± 0.04‰ for Zn. These factors indicate that the heavier isotope preferentially adsorbs onto the oxyhydroxide surface, which is consistent with shorter metal-oxygen bonds and lower coordination number for the metal at the surface relative to the aqueous ion. Fractionation of Cu isotopes also is greater than that for Zn isotopes. Limited isotopic data for adsorption of Cu, Fe(II), and Zn onto amorphous ferric oxyhydroxide suggest that isotopic fractionation is related to the intrinsic equilibrium constants that define aqueous metal interactions with oxyhydroxide surface sites. Greater isotopic fractionation occurs with stronger metal binding by the oxyhydroxide with Cu > Zn > Fe(II).

  17. Common antigen structures of HL-A antigens

    PubMed Central

    Miyakawa, Y.; Tanigaki, N.; Yagi, Y.; Pressman, D.

    1973-01-01

    Antigenic determinants recognizable by rabbits were found to be present on the molecular fragments (48,000 Daltons) which were obtained by papain-solubilization of the membrane fractions of cultured human lymphoid cells and which carried the HL-A determinants. Results were obtained which suggest that these antigenic determinants are present in common on these molecular fragments carrying HL-A determinants regardless of their HL-A specificity and are restricted to the molecular fragments which carry HL-A determinants. The study was made by use of radioimmune methods involving the binding of radioiodine-labelled soluble HL-A antigen preparations by anti-HL-A alloantisera and by rabbit antisera raised against the membrane fractions of cultured human lymphoid cells. PMID:4119543

  18. Protective cellular antigen of Clostridium chauvoei.

    PubMed

    Stevenson, J R; Stonger, K A

    1980-04-01

    Cellular antigens of Clostridium chauvoei, strain IRP-128, were demonstrated to be important in induction of immunity against this bacterium in guinea pigs. At least one major component of the cellular antigen complex was heat-labile. Acid extraction of the bacterial cells, followed by selective purification for flagella, led to the preparation of an acid extract antigen that possessed a high degree of immunogenicity. The acid extract antigen contained flagellar components and was resolved into two major and approximately five minor protein components by polyacrylamide-gel electrophoresis.

  19. Therapeutic effect of ferulic acid, an ethereal fraction of ethanolic extract of seed of Syzygium cumini against streptozotocin-induced diabetes in male rat.

    PubMed

    Mandal, S; Barik, B; Mallick, C; De, D; Ghosh, D

    2008-03-01

    Diabetic therapeutic and antioxidative effects of an ethereal fraction of the ethanolic extract of the seed of Syzygium cumini was studied in streptozotocin (STZ)-induced diabetic rats. Diabetes resulted in a significant elevation in the fasting blood glucose level and in the activity of hepatic glucose-6-phosphatase. There was diminution in the levels of glycogen in the liver and skeletal muscle along with diminution in the activities of hepatic glucose-6-phosphate dehydrogenase, catalase and peroxidase in diabetic rats when compared with controls. Hepatic levels of thiobarbituric acid reactive substance (TBARS) and conjugated dienes (CD) were elevated in respect to control. Oral coadministration of the above fraction to diabetic rats resulted in significant protection in all these parameters. Histological studies of the pancreas showed a qualitative diminution in the area and volume of the islet's of Langerhans, but coadministration of the specific fraction resulted in a significant recovery of the islet's of Langerhans. Chromatography study revealed that the used fraction was ferulic acid (FA). Treatment with FA in normoglycemic rats did not show any significant change in the levels of the selected biosensors. The possible hypothesis for the therapeutic effect of FA against diabetes may be due to its pancreatic beta-cell regenerative effect and/or due to its antioxidant properties.

  20. Changes in the Total Lipid, Neutral Lipid, Phospholipid and Fatty Acid Composition of Phospholipid Fractions during Pastırma Processing, a Dry-Cured Meat Product

    PubMed Central

    Aksu, Muhammet Irfan; Dogan, Mehmet

    2017-01-01

    Pastırma is a dry-cured meat product, produced from whole beef or water buffalo muscles. This study was carried out to investigate the effect of production stages (raw meat, after curing, after 2nd drying and pastırma) on the total lipid, neutral lipid, phospholipid and fatty acid composition of phospholipid fraction of pastırma produced from beef M. Longissimus dorsi muscles. The pH and colour (L*, a* and b*) analyses were also performed in raw meat and pastırma. It was found that pastırma production stages had significant effects (p<0.01) on the total amounts of lipid, neutral lipid and phospholipid, and the highest amounts of lipid, neutral lipid and phospholipid were detected in pastırma. In pastırma, neutral lipid ratio was determined as 79.33±2.06% and phospholipid ratio as 20.67±2.06%. Phospholipids was proportionately lower in pastırma than raw meat. Pastırma production stages affected pentadecanoic acid (15:1) (p<0.01), linoleic acid (18:2n-6) (p<0.05), γ-linoleic acid (18:3n-6) (p<0.05), erucic acid (22:1n-9) (p<0.05), docosapentaenoic acid (22:5n-6) (p<0.05), total unsaturated fatty acid (ΣUSFA) (p<0.05) and total saturated fatty acid (ΣSFA) (p<0.05) ratios of phospholipid fraction and also the moisture content (p<0.01). Pastırma process also affected pH and colour (L*, a* and b*) values (p<0.01), and these values were higher in pastırma than raw meat. PMID:28316467

  1. K99 surface antigen of Escherichia coli: antigenic characterization.

    PubMed Central

    Isaacson, R E

    1978-01-01

    K99 prepared by acid precipitation hemagglutinated guinea pig erythrocytes, whereas K99 prepared by chromatography on diethylaminoethyl-Sephadex did not. K99 purified by either procedure hemagglutinated horse erythrocytes. K99 prepared by acid precipitation contained a second antigen not presnet in the K99 prepared by chromatography on diethylaminoethyl-Sephadex. This antigen could be detected by immunoprecipitation with some, but not all, sera prepared against K99-positive Escherichia coli strains. It was assumed that this second antigen is not K99 and is responsible for the guinea pig erythrocyte hemagglutination reaction. Furthermore, the second antigen has an isoelectric point of 4.2, which has been reported by Morris and co-workers to be the isoelectric point of K99. Images PMID:83300

  2. Characterization of acidic and basic variants of IgG1 therapeutic monoclonal antibodies based on non-denaturing IEF fractionation.

    PubMed

    Dada, Oluwatosin O; Jaya, Nomalie; Valliere-Douglass, John; Salas-Solano, Oscar

    2015-08-20

    Characterization of both the acidic and basic regions of imaged capillary isoelectric focusing (icIEF) profile of an IgG1 antibody was achieved through preparative immobilized pH gradient isoelectric focusing (IPG-IEF) fractionation. Recent attempts at using this method to fractionate charge variants of monoclonal antibodies (mAbs) have shown promising results, but identification of the chemical modifications in the variants was limited to the basic species. We have optimized the method to achieve enrichment of each variant across the icIEF profile of an IgG1 mAb. The fractionation was followed by extended characterization to elucidate the composition of the acidic, main, and basic species observed in the icIEF profile. Deamidation, sialylation, glycation, and fragmentation were identified as the main modifications contributing to acidic variants of the mAb while C-terminal lysine, C-terminal proline amidation, and uncyclized N-terminal glutamine were the major species contributing to the basic variants. This characterization allows a better understanding of the modifications that contribute to the charge variants observed by icIEF, facilitating the evaluation of impacts on product safety and efficacy.

  3. Analysis of heavy oils: Method development and application to Cerro Negro heavy petroleum: Preliminary separation and analysis of acid, base, saturate, and neutral-aromatic fractions

    SciTech Connect

    Green, J.B.; Sturm, G.P. Jr.; Reynolds, J.W.; Thomson, J.S.; Yu, S.K-T.; Grigsby, R.D.; Tang, S.Y.; Shay, J.Y.; Hirsch, D.E.; Sanchez, V.

    1988-10-01

    Cerro Negro 200-425/degree/, 425-550/degree/, and 550-700/degree/C distillates and >700/degree/C residue were separated into acid, base, and neutral concentrates using an improved nonaqueous ion exchange liquid chromatographic procedure. Neutral concentrates were further separated into neutral aromatic and saturated hydrocarbon fractions. A dual column, normal phase high performance liquid chromatographic method was developed for the saturate-aromatic separation. Mass and elemental balances are given for separations of all distillates and the residue. In addition, fractions from the 200-425/degree/C and 425-550/degree/C distillates were analyzed by high resolution mass spectrometry. The applicability of published separation approaches and methods to heavy oil analysis is critically reviewed; the bulk of the available methodology developed for conventional petroleum and synfuels was found to be unproven or unsuitable for heavy oil analysis. Cerro Negro was found to contain 18.2 weight percent acids, 17.6 weight percent bases, 46.9 weight percent neutral aromatics, and 14.7 weight percent saturated hydrocarbons. Saturate fractions contained predominantly cycloparaffins, neutral-aromatics were largely comprised of aromatic hydrocarbons and sulfur compounds, bases were largely nitrogen-containing compounds, and acids were mostly oxygen-containing compounds and nitrogen compounds of pyrrolic type. 145 refs., 24 figs., 21 tabs.

  4. Radioimmunoassays of hidden viral antigens

    SciTech Connect

    Neurath, A.R.; Strick, N.; Baker, L.; Krugman, S.

    1982-07-01

    Antigens corresponding to infectious agents may be present in biological specimens only in a cryptic form bound to antibodies and, thus, may elude detection. We describe a solid-phase technique for separation of antigens from antibodies. Immune complexes are precipitated from serum by polyethylene glycol, dissociated with NaSCN, and adsorbed onto nitrocellulose or polystyrene supports. Antigens remain topographically separated from antibodies after removal of NaSCN and can be detected with radiolabeled antibodies. Genomes from viruses immobilized on nitrocellulose can be identified by nucleic acid hybridization. Nanogram quantities of sequestered hepatitis B surface and core antigens and picogram amounts of hepatitis B virus DNA were detected. Antibody-bound adenovirus, herpesvirus, and measles virus antigens were discerned by the procedure.

  5. Radioimmunoassays of hidden viral antigens.

    PubMed Central

    Neurath, A R; Strick, N; Baker, L; Krugman, S

    1982-01-01

    Antigens corresponding to infectious agents may be present in biological specimens only in a cryptic form bound to antibodies and, thus, may elude detection. We describe a solid phase technique for separation of antigens from antibodies. Immune complexes are precipitated from serum by polyethylene glycol, dissociated with NaSCN, and adsorbed onto nitrocellulose or polystyrene supports. Antigens remain topographically separated from antibodies after removal of NaSCN and can be detected with radiolabeled antibodies. Genomes from viruses immobilized on nitrocellulose can be identified by nucleic acid hybridization. Nanogram quantities of sequestered hepatitis B surface and core antigens and picogram amounts of hepatitis B virus DNA were detected. Antibody-bond adenovirus, herpesvirus, and measles virus antigens were discerned by the procedure. Images PMID:6956871

  6. Isolation and Characterization of Chinese Standard Fulvic Acid Sub-fractions Separated from Forest Soil by Stepwise Elution with Pyrophosphate Buffer

    NASA Astrophysics Data System (ADS)

    Bai, Yingchen; Wu, Fengchang; Xing, Baoshan; Meng, Wei; Shi, Guolan; Ma, Yan; Giesy, John P.

    2015-03-01

    XAD-8 adsorption technique coupled with stepwise elution using pyrophosphate buffers with initial pH values of 3, 5, 7, 9, and 13 was developed to isolate Chinese standard fulvic acid (FA) and then separated the FA into five sub-fractions: FApH3, FApH5, FApH7, FApH9 and FApH13, respectively. Mass percentages of FApH3-FApH13 decreased from 42% to 2.5%, and the recovery ratios ranged from 99.0% to 99.5%. Earlier eluting sub-fractions contained greater proportions of carboxylic groups with greater polarity and molecular mass, and later eluting sub-fractions had greater phenolic and aliphatic content. Protein-like components, as well as amorphous and crystalline poly(methylene)-containing components were enriched using neutral and basic buffers. Three main mechanisms likely affect stepwise elution of humic components from XAD-8 resin with pyrophosphate buffers including: 1) the carboxylic-rich sub-fractions are deprotonated at lower pH values and eluted earlier, while phenolic-rich sub-fractions are deprotonated at greater pH values and eluted later. 2) protein or protein-like components can be desorbed and eluted by use of stepwise elution as progressively greater pH values exceed their isoelectric points. 3) size exclusion affects elution of FA sub-fractions. Successful isolation of FA sub-fractions will benefit exploration of the origin, structure, evolution and the investigation of interactions with environmental contaminants.

  7. Isolation and characterization of Chinese standard fulvic acid sub-fractions separated from forest soil by stepwise elution with pyrophosphate buffer.

    PubMed

    Bai, Yingchen; Wu, Fengchang; Xing, Baoshan; Meng, Wei; Shi, Guolan; Ma, Yan; Giesy, John P

    2015-03-04

    XAD-8 adsorption technique coupled with stepwise elution using pyrophosphate buffers with initial pH values of 3, 5, 7, 9, and 13 was developed to isolate Chinese standard fulvic acid (FA) and then separated the FA into five sub-fractions: FApH3, FApH5, FApH7, FApH9 and FApH13, respectively. Mass percentages of FApH3-FApH13 decreased from 42% to 2.5%, and the recovery ratios ranged from 99.0% to 99.5%. Earlier eluting sub-fractions contained greater proportions of carboxylic groups with greater polarity and molecular mass, and later eluting sub-fractions had greater phenolic and aliphatic content. Protein-like components, as well as amorphous and crystalline poly(methylene)-containing components were enriched using neutral and basic buffers. Three main mechanisms likely affect stepwise elution of humic components from XAD-8 resin with pyrophosphate buffers including: 1) the carboxylic-rich sub-fractions are deprotonated at lower pH values and eluted earlier, while phenolic-rich sub-fractions are deprotonated at greater pH values and eluted later. 2) protein or protein-like components can be desorbed and eluted by use of stepwise elution as progressively greater pH values exceed their isoelectric points. 3) size exclusion affects elution of FA sub-fractions. Successful isolation of FA sub-fractions will benefit exploration of the origin, structure, evolution and the investigation of interactions with environmental contaminants.

  8. Isolation and Characterization of Chinese Standard Fulvic Acid Sub-fractions Separated from Forest Soil by Stepwise Elution with Pyrophosphate Buffer

    PubMed Central

    Bai, Yingchen; Wu, Fengchang; Xing, Baoshan; Meng, Wei; Shi, Guolan; Ma, Yan; Giesy, John P.

    2015-01-01

    XAD-8 adsorption technique coupled with stepwise elution using pyrophosphate buffers with initial pH values of 3, 5, 7, 9, and 13 was developed to isolate Chinese standard fulvic acid (FA) and then separated the FA into five sub-fractions: FApH3, FApH5, FApH7, FApH9 and FApH13, respectively. Mass percentages of FApH3-FApH13 decreased from 42% to 2.5%, and the recovery ratios ranged from 99.0% to 99.5%. Earlier eluting sub-fractions contained greater proportions of carboxylic groups with greater polarity and molecular mass, and later eluting sub-fractions had greater phenolic and aliphatic content. Protein-like components, as well as amorphous and crystalline poly(methylene)-containing components were enriched using neutral and basic buffers. Three main mechanisms likely affect stepwise elution of humic components from XAD-8 resin with pyrophosphate buffers including: 1) the carboxylic-rich sub-fractions are deprotonated at lower pH values and eluted earlier, while phenolic-rich sub-fractions are deprotonated at greater pH values and eluted later. 2) protein or protein-like components can be desorbed and eluted by use of stepwise elution as progressively greater pH values exceed their isoelectric points. 3) size exclusion affects elution of FA sub-fractions. Successful isolation of FA sub-fractions will benefit exploration of the origin, structure, evolution and the investigation of interactions with environmental contaminants. PMID:25735451

  9. Fermentation of alfalfa wet-fractionation liquids to volatile fatty acids by Streptococcus bovis and Megasphaera elsdenii

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A “green juice”, obtained by squeezing freshly harvested alfalfa leaves amended with a commercial lactic acid bacterial inoculant, was readily fermented by 7- to 21-d incubation at room temperature to obtain lactic acid at concentrations of 12-46 g l-1, along with additional acetic and succinic acid...

  10. Immunoreactivity reduction of soybean meal by fermentation, effect on amino acid composition and antigenicity of commercial soy products.

    PubMed

    Song, Y-S; Frias, J; Martinez-Villaluenga, C; Vidal-Valdeverde, C; de Mejia, E Gonzalez

    2008-05-15

    Food allergy has become a public health problem that continues to challenge both the consumer and the food industry. The objectives of this study were to evaluate the reduction of immunoreactivity by natural and induced fermentation of soybean meal (SBM) with Lactobacillus plantarum, Bifidobacterium lactis, Saccharomyces cereviseae, and to assess the effect on amino acid concentration. Immunoreactivity of commercially available fermented soybean products and ingredients was also evaluated. ELISA and western blot were used to measure IgE immunoreactivity using plasma from soy sensitive individuals. Commercial soy products included tempeh, miso and yogurt. Fermented SBM showed reduced immunoreactivity to human plasma, particularly if proteins were <20kDa. S. cereviseae and naturally fermented SBM showed the highest reduction in IgE immunoreactivity, up to 89% and 88%, respectively, against human pooled plasma. When SBM was subjected to fermentation with different microorganisms, most of the total amino acids increased significantly (p<0.05) and only few of them suffered a decrease depending on the type of fermentation. All commercial soy containing products tested showed very low immunoreactivity. Thus, fermentation can decrease soy immunoreactivity and can be optimized to develop nutritious hypoallergenic soy products. However, the clinical relevance of these findings needs to be determined by human challenge studies.

  11. Molecular level description of the sorptive fractionation of a fulvic acid on aluminum oxide using electrospray ionization Fourier transform mass spectrometry.

    PubMed

    Galindo, Catherine; Del Nero, Mirella

    2014-07-01

    We addressed here, by means of electrospray ionization mass spectrometry (ESI-MS) with ultrahigh resolution, the molecular level fractionation of a reference fulvic acid (SRFA) during its sorption at an alumina surface, taken as a model for surfaces of natural aluminum oxide hydrates. Examination of ESI-MS spectra of a native SRFA solution and of supernatants collected in sorption experiments at acidic pH showed that the ∼5700 compounds identified in the native solution were partitioned between the solution and alumina surface to quite varying degrees. Compounds showing the highest affinity for the surface were aromatic compounds with multiple oxygenated functionalities, polycyclic aromatic compounds depleted of hydrogen and carrying few oxygenated groups, and aliphatic compounds with very high O/C values, highlighting the fact that SRFA constituents were sorbed mainly via chemical sorption involving their oxygenated functionalities. We observed an inverse correlation between the degree of sorption of a molecule within a CH2 series and its number of CH2 groups and a positive correlation between the degree of sorption and the number of CO2 groups in a COO series, which was remarkable. These correlations provide evidence at the molecular scale that molecule acidity is the key parameter governing fulvic acid (FA) sorptive fractionation, and they are useful for predicting sorption of FA at a natural oxide surface.

  12. Rat small-intestinal β-galactosidases. Studies on the fractionation of `acid' β-galactosidase with isoelectric focusing, gel filtration and ion-exchange chromatography

    PubMed Central

    Asp, Nils-Georg

    1970-01-01

    1. Different forms of the rat small-intestinal `acid' β-galactosidase were separated by using the isoelectric-focusing technique. The isoelectric points of the different forms were at pH4.2, 4.6, 5.4, 6.1 and approx. 8. 2. The two forms of `acid' β-galactosidase isoelectric at pH4.2 and 4.6 were completely excluded from the Sephadex G-200 gel, whereas the form isoelectric at pH8 had Kav. 0.4. The concentration and pH of the elution buffer influenced the distribution of enzyme activity between different forms. Thus, under certain conditions of ionic strength and pH, the enzyme seems to form high-molecular-weight aggregates with low isoelectric points. These may be homopolymeric aggregates or the result of binding of enzyme to, for example, membrane fragments. The forms isoelectric at pH5.4 and 6.1 are probably aggregates of intermediate size. 3. During ion-exchange chromatography at pH6.0 one fraction of `acid' β-galactosidase was not retained on the column and was isoelectric at pH8 and another fraction was eluted when the buffer concentration in the eluate had increased to about 50mm. The main part of enzyme eluted in this second fraction was also isoelectric at pH8, indicating that the elution of this fraction is not a simple ion-exchange procedure but probably also involves a splitting of high-molecular-weight aggregates, originally retained because of their low isoelectric points. The enzyme subunits have a higher isoelectric point, and are therefore no longer bound to the ion-exchange resin. PMID:5420050

  13. An investigation into the antigenic cross-reactivity of Ophiophagus hannah (king cobra) venom neurotoxin, phospholipase A2, hemorrhagin and L-amino acid oxidase using enzyme-linked immunosorbent assay.

    PubMed

    Tan, N H; Lim, K K; Jaafar, M I

    1993-07-01

    The antigenic cross-reactivity of four Ophiophagus hannah (king cobra) venom components, the neurotoxin (OH-NTX), phospholipase A2 (OH-PLA2), hemorrhagin (OH-HMG) and L-amino acid oxidase (OH-LAAO) were examined by indirect and double sandwich ELISAs. The indirect ELISAs for OH-NTX, OH-PLA2 and OH-HMG were very specific when assayed against the various heterologous snake venoms and O. hannah venom components, at 25 ng/ml antigen level. At higher antigen concentrations (100-400 ng/ml), there were moderate to strong indirect ELISA cross-reactions between anti-O. hannah neurotoxin and venoms from various species of cobra as well as two short neurotoxins. However, anti-O. hannah hemorrhagin did not cross-react with any of the venoms tested, even at these high antigen concentrations, indicating that O. hannah hemorrhagin is antigenically very different from other venom hemorrhagins. Examination of the indirect ELISA cross-reactions between anti-O. hannah PLA2 and several elapid PLA2 enzymes suggests that the elapid PLA2 antigenic class has more than two subgroups. The antibodies to O. hannah L-amino acid oxidase, however, yielded indirect ELISA cross-reactions with many venoms as well as with OH-NTX, OH-PLA2 and OH-HMG, indicating that OH-LAAO shares common epitopes even with unrelated proteins. The double sandwich ELISAs for the four anti-O. hannah venom components, on the other hand, generally exhibited a higher degree of selectivity than the indirect ELISA procedure.

  14. Detection of a low-molecular-weight antigen on melanoma cells by a human antiserum in leukocyte-dependent antibody assays.

    PubMed

    Hersey, P; Murray, E; Werkmeister, J; McCarthy, W H

    1979-10-01

    Biochemical characterization of serologically detected human melanoma antigens was undertaken for the development of immunodiagnostic assays in melanoma. An antiserum from a human melanoma patient, which detected melanoma antigens expressed on a large proportion of different melanoma cells, was used in leucocyte-dependent cytotoxic antibody (LDA) 51Cr-release assays to monitor the purification of melanoma antigens in urea/acetate extracts of lactoperoxidase 125I-labelled melanoma cell membranes. The separation procedures included affinity chromatography on Concanavalin A, gel filtration on porous polyacrylamide beads and preparative isoelectric focusing. The fractions were also monitored by polyacrylamide electrophoresis in sodium dodecyl sulphate and by measurement of beta 2 microglobulin and carcinoembryonic antigen content. The antigens detected by this antiserum appeared to be acidic (pI 3.5) low-mol.-wt glycoproteins of approximately 15,000 daltons which were resistant to heating at 56 degrees C and digestion with neuraminidase, but susceptible to repeated freeze-thawing and trypsin digestion. They did not appear to be related to HLA antigens, beta 2 microglobulin or known foetal antigens. The nature of the antigens detected in these studies is as yet unknown, but they appear similar to those described in the sera and urine of melanoma patients in previous reports. Thes combined results and the frequent expression of these antigens on melanoma cells from different patients suggest that assays to detect this antigen may provide a valuable immunodiagnostic aid in the management of melanoma.

  15. Sodium Taurocholate Modifies the Bile Acid-Independent Fraction of Canalicular Bile Flow in the Rhesus Monkey

    PubMed Central

    Baker, Alfred L.; Wood, R. A. B.; Moossa, A. R.; Boyer, James L.

    1979-01-01

    Bile acid-independent secretion and the choleretic response to taurocholate were determined in rhesus monkeys fitted with indwelling silastic cannulas in the common bile ducts. Bile acids were infused intravenously in random order at 3.5, 7.0, or 10.5 μmol/min for 1.5 h each. When data were analyzed with a single regression line, bile flow increased in proportion to the level of bile acid secretion, although the y-intercepts (the conventional measurement of bile acid-independent secretion) varied widely (77.9±40.9 ml/24 h). The variation in y-intercepts was observed between animals and with repeated studies in the same animal and could not be explained by sex differences or the effects of the indwelling silastic cannulas, but seemed to be related to the order of bile acid infusion. With only two taurocholic acid infusion rates (7.0 and 3.5 μmol/min), [14C]erythritol clearance was greater per mole of secreted bile acid when the initial bile acid infusion was at the high level, but approached zero at low bile acid secretion rates, which suggests that so-called bile acid-independent canalicular flow is closely related to bile acid secretion or is small in size. The augmentation in [14C]erythritol clearance when the high infusion rate was given first was also associated with an increase in biliary clearance of [3H]inulin, which indicates that the premeability to inulin was also enhanced. Identical experiments which substituted equimolar infusions of a nonmicelle-forming bile acid (taurodehydrocholate) for taurocholate failed to demonstrate any difference in choleretic response or biliary clearance of [3H]inulin with the order of bile acid infusion. These experiments demonstrate that a micelleforming bile acid, taurocholate, can increase the permeability of the biliary system to large molecular weight solutes and simultaneously modify the y-intercept and the volume of bile secreted in response to the transported bile acid. Taurocholate may, therefore, modify its own

  16. Carcinoembryonic antigen: assay following heat compared with perchloric acid extraction in patients with colon cancer, non-neoplastic gastrointestinal diseases, or chronic renal failure.

    PubMed

    Witherspoon, L R; Shuler, S E; Alyea, K; Husserl, F E

    1983-10-01

    Heat inactivation has been proposed as an alternative to perchloric acid (PCA) precipitation for the extraction of carcinoembryonic antigen (CEA) from human plasma. We examined a commercial RIA kit using heat inactivation, and compared results with those obtained with PCA precipitation. Adequate sensitivity (1.5 micrograms CEA/l plasma), satisfactory analytical recovery of CEA added to plasma, and dilutional linearity of samples found to have elevated CEA concentrations, were demonstrated for the heat-inactivation assay. Between-assay precision was better with the heat inactivation than with the PCA assay. Although the absolute concentration of CEA estimated after heat inactivation was consistently lower than that estimated after PCA extraction of plasma specimens, there was excellent correlation between results obtained with the two methods in colon cancer patients free of disease, colon cancer patients with residual or recurrent disease, patients with benign gastrointestinal disease, and in patients with chronic renal failure. We conclude that the heat-inactivation assay is an excellent alternative to the PCA assay.

  17. In situ acidity and pH of size-fractionated aerosols during a recent smoke-haze episode in Southeast Asia.

    PubMed

    Behera, Sailesh N; Cheng, Jinping; Balasubramanian, Rajasekhar

    2015-10-01

    The characterization of aerosol acidity has received increased attention in recent years due to its influence on atmospheric visibility, climate change and human health. Distribution of water soluble inorganic (WSI) ions in 12 different size fractions of aerosols was investigated under two different atmospheric conditions (smoke-haze and non-haze periods) in 2012 using the Micro-Orifice Uniform Deposit Impactor (MOUDI) and nano-MOUDI for the first time in Singapore. To estimate the in situ acidity ([H(+)]Ins) and in situ aerosol pH (pHIS), the Aerosol Inorganic Model version-IV under deliquescent mode of airborne particles was used at prevailing ambient temperature and relative humidity. The study revealed an increase in the levels of airborne particulate matter (PM) mass and concentrations of WSI ions for all size fractions during the smoke-haze period, which was caused by the trans-boundary transport of biomass burning-impacted air masses from Indonesia. A bimodal distribution was observed for concentrations of SO4(2-), NO3(-), Cl(-), K(+) and Na(+), whereas concentrations of NH4(+), Ca(2+) and Mg(2+) showed a single mode distribution. The concentration of WSI ions in PM1.8 during the smoke-haze period increased by 3.8 (for SO4(2-)) to 10.5 (for K(+)) times more than those observed during the non-haze period. The pHIS were observed to be lower during the smoke-haze period than that during the non-haze period for all size fractions of PM, indicating that atmospheric aerosols were more acidic due to the influence of biomass burning emissions. The particles in the accumulation mode were more acidic than those in the coarse mode.

  18. Comparable hydrogen isotopic fractionation of plant leaf wax n-alkanoic acids in arid and humid subtropical ecosystems

    NASA Astrophysics Data System (ADS)

    Gao, Li; Zheng, Mei; Fraser, Matthew; Huang, Yongsong

    2014-02-01

    Leaf wax hydrogen isotope proxies have been widely used to reconstruct past hydrological changes. However, published reconstructions have given little consideration for the potentially variable hydrogen isotopic fractionation relative to precipitation (ɛwax-p) under different climate and environmental settings. Chief among various potential factors controlling fractionation is relative humidity, which is known to strongly affect oxygen isotopic ratios of plant cellulose, but its effect on hydrogen isotopic fractionation of leaf waxes is still ambiguous. Analyses of lake surface sediments and individual modern plants have provided valuable information on the variability of ɛwax-p, but both approaches have significant limitations. Here, we present an alternative method to obtain the integrated, time-resolved ecosystem-level ɛwax-p values, by analyzing modern aerosol samples collected weekly from arid (Arizona lowlands) and humid subtropical (Atlanta, Georgia) environments during the main growth season. Because aerosol samples mainly reflect regional leaf wax resources, the extreme contrast in the hydroclimate and associated vegetation assemblages between our study sites allows us to rigorously assess the impact of relative humidity and associated vegetation assemblages on leaf wax hydrogen isotopic fractionation. We show there is only minor difference (mostly <10‰) in the mean ɛwax-p values in the two end-member environments. One possible explanation is that the positive isotopic effects of low relative humidity are offset by progressive replacement of trees with grasses that have a more negative apparent fractionation. Our results represent an important step toward quantitative interpretation of leaf wax hydrogen isotopic records.

  19. Purification and partial characterization of a shed 66 kDa melanoma-associated antigen identified by autologous antibody.

    PubMed

    Vlock, D R; Aul, D J; Toporowicz, A; McCoy, J P; Brown, W E

    1991-10-11

    We have previously reported the isolation of a 66 kDa melanoma-associated antigen, identified by autologous antibody, in serum and unfractionated spent tissue culture media by Western blot analysis. The antigen, detected by autologous serum S150, was found to be broadly represented on melanoma, glioma, renal cell carcinoma, neuroblastoma and head and neck carcinoma cell lines. S150 did not react with bladder or colon carcinoma, fetal fibroblasts, pooled platelets, lymphocytes and red blood cells, autologous cultured lymphocytes or fetal calf serum. To further characterize the antigen, spent tissue culture media, obtained from autologous melanoma cell line, Y-Mel 84:420, was separated by an isoelectric focusing column. Unabsorbed control serum S150 was noted to have a maximum titer of 1:2040 against autologous melanoma cells as measured by protein A hemadsorption. Following isoelectric focusing the greatest decrease in autologous antibody titer (30-fold) occurred with fractions having a pI between 2 and 3. Further resolution of the antigen was accomplished with high-pressure ion-exchange chromatography. One of these fractions showed a significantly higher concentration of antigen and was distinctly resolved from bulk serum albumin. Subsequent Western blot analysis, with autologous antibody, of the isolated antigen-containing fraction, confirmed the presence of a single 66 kDa band. Exposure of the antigen, purified by high-pressure ion-exchange chromatography, to neuraminidase ablated recognition by autologous antibody and suggests that sialic acid is present on the protein and may be part of the antigenic epitope. Binding of antigen, obtained following DEAE anion exchange chromatography, was noted to lectins derived from Triticum vulgaris, Dolichos biflorus and Lycopersicon esculentum. Preparative purification of the antigen was accomplished by anion exchange followed by lectin affinity chromatography with a Dolichos biflorus column. Antigen obtained following

  20. Activity-guided fractionation to characterize a coffee beverage that effectively down-regulates mechanisms of gastric acid secretion as compared to regular coffee.

    PubMed

    Rubach, Malte; Lang, Roman; Skupin, Carola; Hofmann, Thomas; Somoza, Veronika

    2010-04-14

    In some individuals, the consumption of coffee beverages is related to symptoms of gastric irritation. Hot water steam-treatment of raw coffee beans is hypothesized to reduce the contents of stomach irritating compounds, and products to which this technology is applied are launched as stomach-friendly coffee. However, data on the effect of steam-treated coffee on gastric acid secretion are conflicting and it has not been proven yet as to which coffee components act as pro- or antisecretory stimulants. The work presented here aimed at the characterization of a coffee beverage that effectively down-regulates mechanisms of proton secretion in human gastric cells (HGT-1). At first, a regular coffee beverage was fractionated by using solvents of different polarity: water, ethylacetate, dichloromethane, and pentane. Functional assays on the proton secretory activity (PSA) of these solvent fractions revealed the least pronounced effect for the water fraction, for which quantitative analyses demonstrated the highest distribution of chlorogenic acid (95%), (beta)N-alkanoyl-5-hydroxytryptamides (55%), and N-methylpyridinium (N-MP, >99%) among all fractions. Following experiments demonstrated that HGT-1 cells treated with regular coffee fortified with N-MP at a concentration of about 20 mg/mL N-MP showed a significantly decreased PSA as compared to cells which were exposed to coffee beverages containing higher (32-34 mg/L) or lower (5 mg/L) N-MP concentrations. Results from cellular pathway analyses of transcription (ATF-1 and Akt1) and signaling (cAMP and EGFr) factors and kinases (ERK1/2), and experiments on the gene expression of pro (histamine-HRH2 and acetylcholine-CHRM3)- and anti (somatostatin-SSTR1)-secretory receptors and H(+),K(+)-ATPase verified this antisecretory activity of N-MP in coffee beverages.

  1. Total zinc absorption in young women, but not fractional zinc absorption, differs between vegetarian and meat-based diets with equal phytic acid content.

    PubMed

    Kristensen, Mette Bach; Hels, Ole; Morberg, Catrine M; Marving, Jens; Bügel, Susanne; Tetens, Inge

    2006-05-01

    Zn bioavailability is often lower in vegetarian diets mainly due to low Zn and high phytic acid contents. The objective of the present study was to determine the fractional and total absorption of Zn from a vegetarian diet in comparison with meat diets with equal concentrations of phytic acid. A randomized cross-over design, comprising three whole-day diet periods of 5 d each, with a vegetarian diet or diets containing Polish-produced meat or Danish-produced meat, was conducted. Twelve healthy female subjects completed the study. All diets had a high content of phytic acid (1250 micromol/d) and in the meat diets the main meals contained 60 g pork meat. All main meals were extrinsically labelled with the radioactive isotope 65Zn and absorption of Zn was measured in a whole-body counter. The mean Zn content of the whole-day diet was: Polish meat diet 9.9 (SE 0.14) mg, Danish meat diet 9.4 (SE 0.19) mg and vegetarian diet 7.5 (SE 0.18) mg. No difference was observed in the fractional absorption of Zn (Polish meat diet: 27 (SE 1.2) %, Danish meat diet: 27 (SE 1.9) % and vegetarian diet: 23 (SE 2.6) %). A significantly lower amount of total Zn was absorbed from the vegetarian diet (mean Zn absorption of Polish meat diet: 2.7 (SE 0.12) mg/d (P<0.001), Danish meat diet: 2.6 (SE 0.17) mg/d (P=0.006) and vegetarian diet: 1.8 (SE 0.20) mg/d). In conclusion, the vegetarian diet compared with the meat-based diets resulted in lower amounts of absorbed Zn due to a higher content of Zn in the meat diets, but no difference was observed in the fractional absorption of Zn.

  2. Bioconversion of lignocellulosic fraction of water-hyacinth (Eichhornia crassipes) hemicellulose acid hydrolysate to ethanol by Pichia stipitis.

    PubMed

    Kumar, Ashish; Singh, L K; Ghosh, Sanjoy

    2009-07-01

    Fermentation of acid hydrolysate of water-hyacinth (Eichhornia crassipes), a free floating aquatic plant has been investigated for ethanol production. The dilute acid treatment has been applied to utilize the maximum hemicellulosic content of the water-hyacinth. The goal of this work was to investigate, both experimentally and theoretically using mathematical tools, a fermentative system utilizing water-hyacinth (Eichhornia crassipes) hemicellulose acid hydrolysate as a substrate for ethanol production using Pichia stipitis. It was found that 72.83% of xylose was converted to ethanol with a yield of 0.425 g(p)/g(s) and productivity of 0.176 g(p)/L/h. An appropriate mathematical model was developed to explain theoretically the bioconversion of this hemicellulose acid hydrolysate to ethanol and the model was tested statistically to check the validity of the model.

  3. Analysis of Free Drug Fractions in Serum by Ultrafast Affinity Extraction and Two-Dimensional Affinity Chromatography using α1-Acid Glycoprotein Microcolumns

    PubMed Central

    Bi, Cong; Zheng, Xiwei; Hage, David S.

    2016-01-01

    In the circulatory system, many drugs are reversibly bound to serum proteins such as human serum albumin (HSA) and alpha1-acid glycoprotein (AGP), resulting in both free and protein-bound fractions for these drugs. This report examined the use of microcolumns containing immobilized AGP for the measurement of free drug fractions by ultrafast affinity extraction and a two-dimensional affinity system. Several drugs known to bind AGP were used as models to develop and evaluate this approach. Factors considered during the creation of this method included the retention of the drugs on the microcolumns, the injection flow rate, the microcolumn size, and the times at which a second AGP column was placed on-line with the microcolumn. The final system had residence times of only 110–830 ms during sample passage through the AGP microcolumns and allowed free drug fractions to be determined within 10–20 min when using only 3–10 µL of sample per injection. This method was used to measure the free fractions of the model drugs at typical therapeutic levels in serum, giving good agreement with the results obtained by ultrafiltration. This approach was also used to estimate the binding constants for each drug with AGP in serum, even for drugs that had significant interactions with both AGP and HSA in such samples. These results indicated that AGP microcolumns could be used with ultrafast affinity extraction to measure free drug fractions in a label-free manner and to study the binding of drugs with AGP in complex samples such as serum. PMID:26797422

  4. Effects of organic ligands on fractionation of rare earth elements (REEs) in hydroponic plants: an application to the determination of binding capacities by humic acid for modeling.

    PubMed

    Ding, ShiMing; Liang, Tao; Zhang, ChaoSheng; Yan, JunCai; Zhang, ZiLi

    2006-12-01

    Previous studies have revealed the fractionation processes of rare earth elements (REEs) in hydroponic plants, with a heavy REE (HREE, the elements from Gd to Lu) enrichment in leaves. In this study, effects on the HREE enrichment in soybean leaves with additions of carboxylic acids (acetate, malate, citrate, NTA, EDTA and DTPA) and two soil humic acids (HAs) were investigated. REE speciation in carboxylic acid and HA solutions was simulated using Visual MINTEQ and Model V, respectively. The results showed that the effects caused by carboxylic acids were strongly dependent on the differences between their binding strengths for light REEs (LREEs, the elements from La to Eu) and those for HREEs. A good correlation existed between these effects and the changes of free REE ions in solutions. This relationship was also observed for the HA treatments, provided that the intrinsic equilibrium constants of REEs for cation-proton exchange with HA (i.e., pK(MHA)) in Model V were estimated using a free-energy relationship with the stability constants for REE complexation with lactic acid. It is suggested that this set of pK(MHA) values is more suitable for use in Model V for the simulation of REE complexation with HA.

  5. Simultaneous extraction of nucleic acids and proteins from tissue specimens by ultracentrifugation: A protocol using the high-salt protein fraction for quantitative proteome analysis.

    PubMed

    Grzendowski, Michael; Riemenschneider, Markus J; Hawranke, Eva; Stefanski, Anja; Meyer, Helmut E; Reifenberger, Guido; Stühler, Kai

    2009-11-01

    Comprehensive molecular profiling of human tumor tissue specimens at the DNA, mRNA and protein level is often obstructed by a limited amount of available material. Homogenization of frozen tissue samples in guanidine isothiocyanate followed by ultracentrifugation over cesium chloride allows the simultaneous extraction of high-molecular weight DNA and RNA. Here, we present a protocol for quantitative proteome analysis using the high-salt protein fraction obtained as supernatant after ultracentrifugation for nucleic acid extraction. We applied this method to extracts from primary human brain tumors and demonstrate its successful application for protein expression profiling in these tumors using 2-D DIGE, MS and Western blotting.

  6. Selective elimination of the free fatty acid fraction from esterified fatty acids in rat plasma through chemical derivatization and immobilization on amino functionalized silica nano-particles.

    PubMed

    Chen, Jun; Lyu, Qiang; Yang, Mingqing; Chen, Zhi; He, Junhui

    2016-01-29

    A high throughput and low cost approach to separate free fatty acids (FFAs) from phospholipid and acylglycerols (esterified fatty acids, EFAs) has been demonstrated, which may be widely used as a sample preparation method in the metabolomics and lipid research. The optimal conditions for FFAs reacting with N-hydroxysuccinimide (NHS) only need 10min at room temperature to obtain a 93.5% yield of FFAs-NHS ester. The rest 6% FFA transformed into N-cyclohexyl-fatty acid-amide which is stable to methyl esterification adopted for fatty acids analysis. 10min are taken for FFAs-NHS ester to react with amino functionalized silica nanoparticles to immobilize the FFAs. The separation of FFAs from EFAs could be carried out readily by centrifugation. The whole process including derivatization, immobilization, and centrifugation takes less than 40min. Much more accurate fatty acids composition of rat plasma EFAs could be obtained by this approach than the previous reported methods.

  7. Effects of diets containing high or low amounts of stearic acid on plasma lipoprotein fractions and fecal fatty acid excretion of men.

    PubMed

    Dougherty, R M; Allman, M A; Iacono, J M

    1995-05-01

    Ten middle-aged males participated in a crossover study to determine the cholesterolemic effect of high amounts of stearic acid in a natural diet. They consumed a 20-d stabilization diet followed by two 40-d intervention diets containing either 1.5% of energy as stearic (18:0) acid and 7.3% of energy as palmitic (16:0) acid (low stearate: LS) or 2.4% of energy as 16:0 and 7.3% of energy as 18:0 (high stearate: HS). The experimental diets also contained approximately 10% of energy each as saturated and monounsaturated fatty acids and 7.2-8% of energy as polyunsaturated fatty acids. The primary source of 18:0 in the HS diet was sheanut oil (commercially referred to as shea butter) and palm oil and butter in the LS diet. Plasma total, low-density-lipoprotein, and high-density-lipoprotein cholesterol were significantly lower with the HS than with the LS diet. Total fecal fatty acid excretion was higher throughout the HS period. Apparent digestibility of the major dietary fatty acids showed that all of the selected fatty acids, except 18:0, were > or = 95% absorbed. These data demonstrate that feeding diets containing about two times the usual amount of stearic acid consumed in the United States, contributed to an increase in plasma lipoprotein concentrations at 40 d from an earlier decrease at 20 d. The time required to achieve stable cholesterol concentrations appears to vary depending on the kind of saturated fatty acids present in the diet.

  8. Interactive effects of elevated CO2 and nitrogen deposition on fatty acid molecular and isotope composition of above- and belowground tree biomass and forest soil fractions.

    PubMed

    Griepentrog, Marco; Eglinton, Timothy I; Hagedorn, Frank; Schmidt, Michael W I; Wiesenberg, Guido L B

    2015-01-01

    Atmospheric carbon dioxide (CO2) and reactive nitrogen (N) concentrations have been increasing due to human activities and impact the global carbon (C) cycle by affecting plant photosynthesis and decomposition processes in soil. Large amounts of C are stored in plants and soils, but the mechanisms behind the stabilization of plant- and microbial-derived organic matter (OM) in soils are still under debate and it is not clear how N deposition affects soil OM dynamics. Here, we studied the effects of 4 years of elevated (13C-depleted) CO2 and N deposition in forest ecosystems established in open-top chambers on composition and turnover of fatty acids (FAs) in plants and soils. FAs served as biomarkers for plant- and microbial-derived OM in soil density fractions. We analyzed above- and belowground plant biomass of beech and spruce trees as well as soil density fractions for the total organic C and FA molecular and isotope (δ13C) composition. FAs did not accumulate relative to total organic C in fine mineral fractions, showing that FAs are not effectively stabilized by association with soil minerals. The δ13C values of FAs in plant biomass increased under high N deposition. However, the N effect was only apparent under elevated CO2 suggesting a N limitation of the system. In soil fractions, only isotope compositions of short-chain FAs (C16+18) were affected. Fractions of 'new' (experimental-derived) FAs were calculated using isotope depletion in elevated CO2 plots and decreased from free light to fine mineral fractions. 'New' FAs were higher in short-chain compared to long-chain FAs (C20-30), indicating a faster turnover of short-chain compared to long-chain FAs. Increased N deposition did not significantly affect the quantity of 'new' FAs in soil fractions, but showed a tendency of increased amounts of 'old' (pre-experimental) C suggesting that decomposition of 'old' C is retarded by high N inputs.

  9. Bacillus subtilis-specific poly-gamma-glutamic acid regulates development pathways of naive CD4(+) T cells through antigen-presenting cell-dependent and -independent mechanisms.

    PubMed

    Kim, Sunghoon; Yang, Jun Young; Lee, Kyuheon; Oh, Kyu Heon; Gi, Mia; Kim, Jung Mogg; Paik, Doo Jin; Hong, Seokmann; Youn, Jeehee

    2009-08-01

    Peripheral naive CD4(+) T cells selectively differentiate to type 1 T(h), type 2 T(h) and IL-17-producing T(h) (T(h)17) cells, depending on the priming conditions. Since these subsets develop antagonistically to each other to elicit subset-specific adaptive immune responses, balance between these subsets can regulate the susceptibility to diverse immune diseases. The present study was undertaken to determine whether poly-gamma-glutamic acid (gamma-PGA), an edible and safe exopolymer that is generated by microorganisms such as Bacillus subtilis, could modulate the development pathways of T(h) subsets. The presence of gamma-PGA during priming promoted the development of T(h)1 and T(h)17 cells but inhibited development of T(h)2 cells. gamma-PGA up-regulated the expression of T-bet and ROR-gammat, the master genes of T(h)1 and T(h)17 cells, respectively, whereas down-regulating the level of GATA-3, the master gene of T(h)2 cells. gamma-PGA induced the expression of IL-12p40, CD80 and CD86 in dendritic cells (DC) and macrophages in a Toll-like receptor-4-dependent manner, and the effect of gamma-PGA on T(h)1/T(h)2 development was dependent on the presence of antigen-presenting cells (APC). Furthermore, gamma-PGA-stimulated DC favored the polarization of naive CD4(+) T cells toward T(h)1 cells rather than T(h)2 cells. In contrast, gamma-PGA affected T(h)17 cell development, regardless of the presence or absence of APC. Thus, these data demonstrate that gamma-PGA has the potential to regulate the development pathways of naive CD4(+) T cells through APC-dependent and -independent mechanisms and to be applicable to treating T(h)2-dominated diseases.

  10. Fractionation of iron isotopes during leaching of natural particles by acidic and circumneutral leaches and development of an optimal leach for marine particulate iron isotopes

    NASA Astrophysics Data System (ADS)

    Revels, Brandi N.; Zhang, Ruifeng; Adkins, Jess F.; John, Seth G.

    2015-10-01

    Iron (Fe) is an essential nutrient for life on land and in the oceans. Iron stable isotope ratios (δ56Fe) can be used to study the biogeochemical cycling of Fe between particulate and dissolved phases in terrestrial and marine environments. We have investigated the dissolution of Fe from natural particles both to understand the mechanisms of Fe dissolution, and to choose a leach appropriate for extracting labile Fe phases of marine particles. With a goal of finding leaches which would be appropriate for studying dissolved-particle interactions in an oxic water column, three particle types were chosen including oxic seafloor sediments (MESS-3), terrestrial dust (Arizona Test Dust - A2 Fine), and ocean sediment trap material from the Cariaco basin. Four leaches were tested, including three acidic leaches similar to leaches previously applied to marine particles and sediments (25% acetic acid, 0.01 N HCl, and 0.5 N HCl) and a pH 8 oxalate-EDTA leach meant to mimic the dissolution of particles by organic complexation, as occurs in natural seawater. Each leach was applied for three different times (10 min, 2 h, 24 h) at three different temperatures (25 °C, 60 °C, 90 °C). MESS-3 was also leached under various redox conditions (0.02 M hydroxylamine hydrochloride or 0.02 M hydrogen peroxide). For all three sample types tested, we find a consistent relationship between the amount of Fe leached and leachate δ56Fe for all of the acidic leaches, and a different relationship between the amount of Fe leached and leachate δ56Fe for the oxalate-EDTA leach, suggesting that Fe was released through proton-promoted dissolution for all acidic leaches and by ligand-promoted dissolution for the oxalate-EDTA leach. Fe isotope fractionations of up to 2‰ were observed during acidic leaching of MESS-3 and Cariaco sediment trap material, but not for Arizona Test Dust, suggesting that sample composition influences fractionation, perhaps because Fe isotopes are greatly fractionated

  11. Effects of Low-Molecular-Weight Organic Acids on the Dissolution of Hydroxyapatite Nanoparticles in Batch and Column Experiments: A Perspective from Phosphate Oxygen Isotope Fractionation

    NASA Astrophysics Data System (ADS)

    Wang, D.; Jaisi, D. P.; Jin, Y.

    2015-12-01

    Hydroxyapatite nanoparticles (HANPs) are increasingly being advocated as an efficient and environment-friendly "green" phosphorus nanofertilizer attributed to their nanoscale dimension, large reactive surface area, and low leaching potential. However, knowledge of how naturally occurring low-molecular-weight organic acids (LMWOAs) that are secreted by plant roots mediate the dissolution of HANPs (releasing PO43- ion for plant growth) is nonexistent. Here three most commonly encountered LMWOAs (acetic acid, oxalic acid, and citric acid) at environmentally relevant concentration (1 mM) were evaluated for their effects on HANPs' dissolution in static batch and dynamic column systems. Particularly, phosphate oxygen isotope fractionation of HANPs during dissolution was examined to disentangle mechanisms controlling the evolution of O-isotopic composition of dissolved PO43- ion. Our results reveal that in batch experiments the dissolution of HANPs was fast but the overall dissolution efficiency of HANPs was limited (≤30%). In contrast, ~100% HANPs were dissolved in columns where LMWOAs were continuously injected. The limited dissolution of HANPs in static batch systems was due primarily to pH buffer effect (pH increased sharply when LMWOA was added in HANPs suspension), whereas in dynamic column systems the HANPs were continuously dissolved by low pH LMWOAs and leached away. Regardless of LMWOA type and experimental system, the isotopically light phosphate (P16O4) was preferentially released during dissolution and the O-isotopic composition of dissolved PO43- ion increased gradually with increasing dissolution due to equilibrium isotope effect between dissolved PO43- ion and HANPs. However, the overall magnitude of O-isotopic fractionation of dissolved PO43- ion was less in batch than in column systems, due to less mass transfer between dissolved PO43- ions and HANPs in batch relative to column experiments. Our findings provide new insights into bioavailability

  12. Isotopic composition of Murchison organic compounds: Intramolecular carbon isotope fractionation of acetic acid. Simulation studies of cosmochemical organic syntheses

    NASA Technical Reports Server (NTRS)

    Yuen, G. U.; Cronin, J. R.; Blair, N. E.; Desmarais, D. J.; Chang, S.

    1991-01-01

    Recently, in our laboratories, samples of Murchison acetic acid were decarboxylated successfully and the carbon isotopic composition was measured for the methane released by this procedure. These analyses showed significant differences in C-13/C-12 ratios for the methyl and carboxyl carbons of the acetic acid molecule, strongly suggesting that more than one carbon source may be involved in the synthesis of the Murchison organic compounds. On the basis of this finding, laboratory model systems simulating cosmochemical synthesis are being studied, especially those processes capable of involving two or more starting carbon sources.

  13. Toxicity of oil sands acid-extractable organic fractions to freshwater fish: Pimephales promelas (fathead minnow) and Oryzias latipes (Japanese medaka).

    PubMed

    Bauer, Anthony E; Frank, Richard A; Headley, John V; Peru, Kerry M; Farwell, Andrea J; Dixon, D George

    2017-03-01

    The Alberta oil sands are one of the largest global petroleum deposits and, due to non-release practices for oil sands process-affected waters, produced tailings are stored in large ponds. The acid extractable organic (AEO) compounds in oil sands process-affected water are of greatest concern due to their persistence and toxicity to a variety of aquatic biota. The present study evaluated the toxicity of the five AEO fractions to two fish species: Oryzias latipes (Japanese medaka) and Pimephales promelas (fathead minnow). The fractions (F1-F5) were comprised of AEO with increasing mean molecular weight and subsequent increases in cyclicity, aromaticity, degree of oxygenation, and heteroatom content. The lowest molecular weight fraction, F1, displayed the lowest acute toxicity to both fish species. For fathead minnow, F5 displayed the greatest toxic potency, while F2 to F4 displayed intermediate toxicities. For Japanese medaka, F2 and F3 displayed the greatest acute toxicities and F1, F4 and F5 were significantly less potent. Overall, fathead minnow were more acutely sensitive to AEO than Japanese medaka. The present study indicates that AEO toxicity may not be solely driven by a narcotic mode of action, but chemical composition such as aromaticity and heteroatom content and their relation to toxicity suggest other drivers indicative of additional modes of toxic action.

  14. Antigenic determinants and functional domains in core antigen and e antigen from hepatitis B virus.

    PubMed Central

    Salfeld, J; Pfaff, E; Noah, M; Schaller, H

    1989-01-01

    The precore/core gene of hepatitis B virus directs the synthesis of two polypeptides, the 21-kilodalton subunit (p21c) forming the viral nucleocapsid (serologically defined as core antigen [HBcAg]) and a secreted processed protein (p17e, serologically defined as HBe antigen [HBeAg]). Although most of their primary amino acid sequences are identical, HBcAg and HBeAg display different antigenic properties that are widely used in hepatitis B virus diagnosis. To locate and to characterize the corresponding determinants, segments of the core gene were expressed in Escherichia coli and probed with a panel of polyclonal or monoclonal antibodies in radioimmunoassays or enzyme-linked immunosorbent assays, Western blots, and competition assays. Three distinct major determinants were characterized. The single conformational determinant responsible for HBc antigenicity in the assembled core (HBc) and a linear HBe-related determinant (HBe1) were both mapped to an overlapping hydrophilic sequence around amino acid 80; a second HBe determinant (HBe2) was assigned to a location in the vicinity of amino acid 138 but found to require for its antigenicity the intramolecular participation of the extended sequence between amino acids 10 and 140. It is postulated that HBcAg and HBeAg share common basic three-dimensional structure exposing the common linear determinant HBe1 but that they differ in the presentation of two conformational determinants that are either introduced (HBc) or masked (HBe2) in the assembled core. The simultaneous presentation of HBe1 and HBc, two distinctly different antigenic determinants with overlapping amino acid sequences, is interpreted to indicate the presence of slightly differently folded, stable conformational states of p21c in the hepatitis B virus nucleocapsid. Images PMID:2463383

  15. Lipids and Fatty Acids in Algae: Extraction, Fractionation into Lipid Classes, and Analysis by Gas Chromatography Coupled with Flame Ionization Detector (GC-FID).

    PubMed

    Guihéneuf, Freddy; Schmid, Matthias; Stengel, Dagmar B

    2015-01-01

    Despite the number of biochemical studies exploring algal lipids and fatty acid biosynthesis pathways and profiles, analytical methods used by phycologists for this purpose are often diverse and incompletely described. Potential confusion and potential variability of the results between studies can therefore occur due to change of protocols for lipid extraction and fractionation, as well as fatty acid methyl esters (FAME) preparation before gas chromatography (GC) analyses. Here, we describe a step-by-step procedure for the profiling of neutral and polar lipids using techniques such as solid-liquid extraction (SLE), thin-layer chromatography (TLC), and gas chromatography coupled with flame ionization detector (GC-FID). As an example, in this protocol chapter, analyses of neutral and polar lipids from the marine microalga Pavlova lutheri (an EPA/DHA-rich haptophyte) will be outlined to describe the distribution of fatty acid residues within its major lipid classes. This method has been proven to be a reliable technique to assess changes in lipid and fatty acid profiles in several other microalgal species and seaweeds.

  16. Production of furfural from xylose, water-insoluble hemicelluloses and water-soluble fraction of corncob via a tin-loaded montmorillonite solid acid catalyst.

    PubMed

    Li, Huiling; Ren, Junli; Zhong, Linjie; Sun, Runcang; Liang, Lei

    2015-01-01

    The conversion of xylose, water-insoluble hemicelluloses (WIH) and water-soluble fraction (WSF) of corncob to furfural was performed using montmorillonite with tin ions (Sn-MMT) containing double acid sites as a solid acid catalyst. The co-existence of Lewis acids and Brønsted acids in Sn-MMT was shown to improve the furfural yield and selectivity. 76.79% furfural yield and 82.45% furfural selectivity were obtained from xylose using Sn-MMT as a catalyst in a biphasic system with 2-s-butylphenol (SBP) as the organic extracting layer and dimethyl sulfoxide (DMSO) as the co-solvent in contact with an aqueous phase saturated with NaCl (SBP/NaCl-DMSO) at 180°C for 30min. Furthermore, Sn-MMT also demonstrated the excellent catalytic performance in the conversion of pentose-rich materials of corncob and 39.56% and 54.15% furfural yields can be directly obtained from WIH and WSF in the SBP/NaCl-DMSO system, respectively.

  17. Purification of human seminal plasma no. 7 antigen by immunoaffinity chromatography on bound monoclonal antibody.

    PubMed Central

    Isojima, S; Koyama, K; Fujiwara, N

    1982-01-01

    Human seminal plasma (HSP) No. 7 antigen was purified by immunoaffinity chromatography on bound 1C4 monoclonal antibody (Moab) (Shigeta et al., 1980b). The pooled HSP protein was applied to a CNBr-activated Sepharose 4B column of bound 1C4 Moab gamma globulin and the antibody bound fraction (fr) eluted was further purified by rechromatography in the same way. The purified antigen in the antibody bound fr obtained by rechromatography gave a single band on SDS-PAGE in a position corresponding to a molecular weight of 15,000 daltons. This preparation was 196.2 times more effective than the original HSP protein in neutralizing the sperm immobilizing activity of 1C4 Moab. The purified HSP No. 7 antigen contained iron, but was different from lactoferrin and transferrin. It did not show any enzymatic activities, such as those of acid phosphatase, LDH or trypsin inhibitor, and shared antigenicity with human milk protein. It was present in seminal plasma as a molecule with a higher molecular weight but seemed to be cleaved to a monomer of 15,000 daltons during purification procedures. This antigen is present on spermatozoa as sperm-coating antigen and the corresponding antibody can immobilize spermatozoa with complement. Images Fig. 3 PMID:7127911

  18. Separation of soluble Brucella antigens by gel-filtration chromatography.

    PubMed

    McGhee, J R; Freeman, B A

    1970-07-01

    Soluble precipitating antigens of Brucella suis have been, in various degrees, purified by filtration on Sephadex gels. The most useful gels employed were Sephadex G-150, Sephadex G-200, and Sepharose 4B. Although not all fractions proved to be immunologically pure, some crude molecular-size estimates of most of the 13 soluble antigens of the Brucella cell could be given. In addition, monospecific antisera to three purified Brucella antigens have been prepared. By using purified preparations, physical and chemical data were obtained on two major antigens, E and 1, and a minor antigen, f. Antigen E is not an agglutinogen and may be toxic. Antigen 1 is of low molecular weight and is neither toxic nor agglutinogenic. The minor antigen f is an agglutinogen as well as a precipitinogen and is found on the cell surface. Both major antigens, when purified, were immunogenic in rabbits.

  19. Fractionated illumination after topical application of 5-aminolevulinic acid on normal skin of hairless mice: the influence of the dark interval.

    PubMed

    de Bruijn, H S; van der Ploeg-van den Heuvel, A; Sterenborg, H J C M; Robinson, D J

    2006-12-01

    We have previously shown that light fractionation during topical aminolevulinic acid based photodynamic therapy (ALA-PDT) with a dark interval of 2h leads to a significant increase in efficacy in both pre-clinical and clinical PDT. However this fractionated illumination scheme required an extended overall treatment time. Therefore we investigated the relationship between the dark interval and PDT response with the aim of reducing the overall treatment time without reducing the efficacy. Five groups of mice were treated with ALA-PDT using a single light fraction or the two-fold illumination scheme with a dark interval of 30 min, 1, 1.5 and 2h. Protoporphyrin IX fluorescence kinetics were monitored during illumination. Visual skin response was monitored in the first seven days after PDT and assessed as PDT response. The PDT response decreases with decreasing length of the dark interval. Only the dark interval of 2h showed significantly more damage compared to all the other dark intervals investigated (P<0.05 compared to 1.5h and P<0.01 compared to 1h, 30 min and a single illumination). No relationship could be shown between the utilized PpIX fluorescence during the two-fold illumination and the PDT response. The rate of photobleaching was comparable for the first and the second light fraction and not dependent of the length of dark interval used. We conclude that in the skin of the hairless mouse the dark interval cannot be reduced below 2h without a significant reduction in PDT efficacy.

  20. Bulk vs. amino acid stable N isotope estimations of metabolic status and contributions of nitrogen fixation to size-fractionated zooplankton biomass in the subtropical N Atlantic

    NASA Astrophysics Data System (ADS)

    Mompeán, Carmen; Bode, Antonio; Gier, Elizabeth; McCarthy, Matthew D.

    2016-08-01

    A comparative analysis of natural abundance of stable N isotopes (δ15N) in individual amino acids and bulk organic matter of size-fractionated plankton revealed the differential impact of nitrogen fixation through the food web in a transect across the subtropical North Atlantic. All δ15N measurements showed low values in the central region, followed by the western zone, while maximum δ15N values were found in the eastern zone. These results were consistent with the prevalence of nitrogen fixation in the central and western zones, and the influence of the west Africa upwelling in the eastern zone. Use of compound-specific amino acid isotope data (CSI-AA) revealed relatively low variability in the impact of diazotrophic nitrogen within the different plankton size fractions, while δ15N of bulk organic matter showed high variability with size. Explicit CSI-AA trophic position estimates showed a small increase with mean plankton size class and varied in a relatively narrow range 1.8-2.5), with the lowest values in the central zone. High correlations between bulk plankton δ15N and individual amino acids (in particular Phe and Thr), as well as reconstructed total protein δ15N values, suggest a set of new relationships that may be important to tracing direct plankton contributions to nitrogen recycling in the ocean, including detrital organic nitrogen pools. Overall, these new results represent the most detailed investigation of CSI-AA data in plankton size classes to date, and indicated a greater importance of diazotrophic N than suggested by concurrent measurements of bulk δ15N, abundance of large nitrogen fixing organisms or nitrogen fixation rates.

  1. Fractional randomness

    NASA Astrophysics Data System (ADS)

    Tapiero, Charles S.; Vallois, Pierre

    2016-11-01

    The premise of this paper is that a fractional probability distribution is based on fractional operators and the fractional (Hurst) index used that alters the classical setting of random variables. For example, a random variable defined by its density function might not have a fractional density function defined in its conventional sense. Practically, it implies that a distribution's granularity defined by a fractional kernel may have properties that differ due to the fractional index used and the fractional calculus applied to define it. The purpose of this paper is to consider an application of fractional calculus to define the fractional density function of a random variable. In addition, we provide and prove a number of results, defining the functional forms of these distributions as well as their existence. In particular, we define fractional probability distributions for increasing and decreasing functions that are right continuous. Examples are used to motivate the usefulness of a statistical approach to fractional calculus and its application to economic and financial problems. In conclusion, this paper is a preliminary attempt to construct statistical fractional models. Due to the breadth and the extent of such problems, this paper may be considered as an initial attempt to do so.

  2. Isolation and characterization of a human liver and kidney-specific protein: the hepato-renal (H-R) antigen.

    PubMed Central

    Nerenberg, S T; Prasad, R; Inboriboon, P; Biskup, N; Pedersen, L; Faiferman, I

    1980-01-01

    This paper reports the isolation and characterization of a soluble antigen shared by the liver and kidney of human and some other animal species. Homogenates of human liver in saline were centrifugated at 27,000 g and the supernatants were fractionated by preparative polyacrylamide gel electrophoresis. The gels were divided in sections and each was injected into rabbits; after absorption with polymerized normal human serum, the antiserum obtained by injecting one of the sections reacted only with saline extracts of human liver and kidney when tested against a variety of human tissue extracts. The absorbed antiserum, polymerized and insolubilized with glutaraldehyde, was used to purify the antigen by affinity chromatography. The purified antigen proved to be a glycoprotein containing 19 percent carbohydrate, had a molecular weight of 5.8-6.0 x 10(4) Daltons and a pI of 7.2-7.4. The antigen, relatively thermostable, was precipitated by 35-55 percent ammonium sulphate; its antigenic activity was not affected by extraction with 0.6 N perchloric acid or by incubation with ribonuclease, deoxyribonuclease or neuraminidase but was destroyed by incubation with ttypsin or chymotrypsin. Immunoperoxidase studies showed that the antigen appeared concentrated in the neclei of liver and kidney glomerular epithelial and tubular epithelial cells in humans and rats. The antigen could not be detected in human hepatomas or hypernephromas or in the rat Morris hepatoma 5123. Images FIG. 1 FIG. 7 PMID:6155231

  3. Natural selection promotes antigenic evolvability.

    PubMed

    Graves, Christopher J; Ros, Vera I D; Stevenson, Brian; Sniegowski, Paul D; Brisson, Dustin

    2013-01-01

    The hypothesis that evolvability - the capacity to evolve by natural selection - is itself the object of natural selection is highly intriguing but remains controversial due in large part to a paucity of direct experimental evidence. The antigenic variation mechanisms of microbial pathogens provide an experimentally tractable system to test whether natural selection has favored mechanisms that increase evolvability. Many antigenic variation systems consist of paralogous unexpressed 'cassettes' that recombine into an expression site to rapidly alter the expressed protein. Importantly, the magnitude of antigenic change is a function of the genetic diversity among the unexpressed cassettes. Thus, evidence that selection favors among-cassette diversity is direct evidence that natural selection promotes antigenic evolvability. We used the Lyme disease bacterium, Borrelia burgdorferi, as a model to test the prediction that natural selection favors amino acid diversity among unexpressed vls cassettes and thereby promotes evolvability in a primary surface antigen, VlsE. The hypothesis that diversity among vls cassettes is favored by natural selection was supported in each B. burgdorferi strain analyzed using both classical (dN/dS ratios) and Bayesian population genetic analyses of genetic sequence data. This hypothesis was also supported by the conservation of highly mutable tandem-repeat structures across B. burgdorferi strains despite a near complete absence of sequence conservation. Diversification among vls cassettes due to natural selection and mutable repeat structures promotes long-term antigenic evolvability of VlsE. These findings provide a direct demonstration that molecular mechanisms that enhance evolvability of surface antigens are an evolutionary adaptation. The molecular evolutionary processes identified here can serve as a model for the evolution of antigenic evolvability in many pathogens which utilize similar strategies to establish chronic infections.

  4. Ethanol production from residual wood chips of cellulose industry: acid pretreatment investigation, hemicellulosic hydrolysate fermentation, and remaining solid fraction fermentation by SSF process.

    PubMed

    Silva, Neumara Luci Conceição; Betancur, Gabriel Jaime Vargas; Vasquez, Mariana Peñuela; Gomes, Edelvio de Barros; Pereira, Nei

    2011-04-01

    Current research indicates the ethanol fuel production from lignocellulosic materials, such as residual wood chips from the cellulose industry, as new emerging technology. This work aimed at evaluating the ethanol production from hemicellulose of eucalyptus chips by diluted acid pretreatment and the subsequent fermentation of the generated hydrolysate by a flocculating strain of Pichia stipitis. The remaining solid fraction generated after pretreatment was subjected to enzymatic hydrolysis, which was carried out simultaneously with glucose fermentation [saccharification and fermentation (SSF) process] using a strain of Saccharomyces cerevisiae. The acid pretreatment was evaluated using a central composite design for sulfuric acid concentration (1.0-4.0 v/v) and solid to liquid ratio (1:2-1:4, grams to milliliter) as independent variables. A maximum xylose concentration of 50 g/L was obtained in the hemicellulosic hydrolysate. The fermentation of hemicellulosic hydrolysate and the SSF process were performed in bioreactors and the final ethanol concentrations of 15.3 g/L and 28.7 g/L were obtained, respectively.

  5. Advances in mass spectrometric characterization of naphthenic acids fraction compounds in oil sands environmental samples and crude oil--A review.

    PubMed

    Headley, John V; Peru, Kerry M; Barrow, Mark P

    2016-01-01

    There has been a recent surge in the development of mass spectrometric methods for detailed characterization of naphthenic acid fraction compounds (all C(c)H(h)N(n)O(o)S(s), species, including heteroatomic and aromatic components in the acid-extractable fraction) in environmental samples. This surge is driven by the increased activity in oil sands environmental monitoring programs in Canada, the exponential increase in research studies on the isolation and toxicity identification of components in oil sands process water (OSPW), and the analytical requirements for development of technologies for treatment of OSPW. There has been additional impetus due to the parallel studies to control corrosion from naphthenic acids during the mining and refining of heavy bitumen and crude oils. As a result, a range of new mass spectrometry tools have been introduced since our last major review of this topic in 2009. Of particular significance are the developments of combined mass spectrometric methods that incorporate technologies such as gas chromatography, liquid chromatography, and ion mobility. There has been additional progress with respect to improved visualization methods for petroleomics and oil sands environmental forensics. For comprehensive coverage and more reliable characterization of samples, an approach based on multiple-methods that employ two or more ionization modes is recommended. On-line or off-line fractionation of isolated extracts, with or without derivatization, might also be used prior to mass spectrometric analyses. Individual ionization methods have their associated strengths and weaknesses, including biases, and thus dependence upon a single ionization method is potentially misleading. There is also a growing trend to not rely solely on low-resolution mass spectrometric methods (<20,000 resolving power at m/z 200) for characterization of complex samples. Future research is anticipated to focus upon (i) structural elucidation of components to determine

  6. Relationship of spermatoscopy, prostatic acid phosphatase activity and prostate-specific antigen (p30) assays with further DNA typing in forensic samples from rape cases.

    PubMed

    Romero-Montoya, Lydia; Martínez-Rodríguez, Hugo; Pérez, Miguel Antonio; Argüello-García, Raúl

    2011-03-20

    In the forensic laboratory the biological analyses for rape investigation commonly include vaginal swabs as sample material combined to biochemical tests including sperm cytology (SC) and detection of acid phosphatase activity (AP) and prostate-specific antigen (PSA, p30) for the conclusive identification of semen components. Most reports comparing these tests relied on analysis of semen samples or donor swabs taken under controlled conditions; however their individual or combined efficacy under real live sampling conditions in different laboratories is largely unknown. We carried out SC, APA and PSA analyses in vaginal swabs collected from casework rapes submitted to Mexican Forensic Laboratories at Texcoco and Toluca. On the basis of positive and negative results from each assay and sample, data were classified into eight categories (I-VIII) and compared with those obtained in the two only similar studies reported in Toronto, Canada and Hong Kong, China. SC and APA assays had the higher overall positivity in Toluca and Texcoco samples respectively and otherwise PSA had a lower but very similar positivity between these two laboratories. When compared to the previous studies some similarities were found, namely similar frequencies (at a ratio of approximately 1 out of 3) of samples being positive or negative by all techniques (Categories I and VI respectively) and a comparable overall positivity of APA and SC but higher than that of PSA. Indeed the combined results of using SC, APA and PSA tests was considered as conclusive for semen detection from approximately 1 out of 3 cases (Category I) to approximately 1 out of 2 cases in a scenario where at least SC is positive, strongly presumptive in 2 out of 3 cases (with at least one test positive) and the remainder 1 out of 3 cases (Category VI) suggested absence of semen. By determining Y-STR polymorphisms (12-loci) in additional samples obtained at Toluca laboratory, complete DNA profiles were determined from all

  7. Formation mechanisms of monochloropropanediol (MCPD) fatty acid diesters in refined palm (Elaeis guineensis) oil and related fractions.

    PubMed

    Destaillats, Frédéric; Craft, Brian D; Sandoz, Laurence; Nagy, Kornél

    2012-01-01

    Monochloropropanediol (MCPD) fatty acid esters are process contaminants generated during the deodorisation of edible oils. In particular, MCPD diesters are found in higher abundance in refined palm oil than other edible oils. In the present study, a series of model reactions mimicking palm oil deodorisation has been conducted with pure acylglycerols in the presence or absence of either organic or inorganic chlorine-containing compounds. Results showed that the bulk of MCPD diesters are formed above 200°C through the reaction of organochlorines with triacylglycerols (TAG). Additional experiments confirmed that this reaction can be initiated during palm oil deodorisation by hydrogen chloride (HCl) gas evolved through the thermal degradation of organochlorines present in the oil. Therein, the majority of the ultimately produced MCPD diesters are the result of HCl reacting with TAG, via protonation, followed by the elimination of a fatty acid residue. Two possible MCPD diester formation mechanisms are highlighted, both of which involve acyloxonium ion reactive intermediates. Investigations with pure TAG regio-isomers showed that MCPD ester formation is regioselective and the sn-1(3) position of the glycerol backbone is favoured.

  8. Aluminium (Al) fractionation and speciation; getting closer to describing the factors influencing Al(3+) in water impacted by acid mine drainage.

    PubMed

    Chamier, Jessica; Wicht, Merrill; Cyster, Lilburne; Ndindi, Nosintu P

    2015-07-01

    Acid mine drainage (AMD) severely impacts the water chemistry of a receiving resource, changing the occurrence, speciation and toxicity of metals such as Aluminium (Al). The toxicity of Al is determined by its speciation represented by the labile monomer Al fraction or Al(3+). The purpose of the study was to combine fractionation and Visual MINTEQ speciation to calculate the effect of AMD altered water chemistry on Al speciation and Al(3+) concentration. Water in rivers impacted by AMD presented with monomeric Al (Almon) concentrations between 0.35 and 15.37mgL(-)(1) which existed almost exclusively in the toxic labile form (98%). For the reference site, Almon was less than 2% (10μgL(-1)), suggesting significantly lower Al toxicity. Principal component analysis plots illustrated that labile Al was directly related to the total Al and iron concentrations and strongly influenced by parameters such as pH, electrical conductivity, sulphate and dissolved organic carbon. Visual MINTEQ modelling was used to determine the primary Al species distribution. The dominant form of Al in AMD impacted water was AlSO4(+), which increased proportionally with the sulphate and Al(3+) concentration. Heavily impacted areas, presented with an average of 1mgmL(-)(1) Al(3+), which poses a potential human health risk. A novel centrifugal ultrafiltration method was investigated as an alternative to determining Almon to simplify the speciation of Al. Monomeric and centrifugal ultrafiltrated (<10kD) Al fractions were significantly similar (p=0.74), suggesting that ultrafiltration may present a time, energy and cost saving alternative to organic extraction of Almon.

  9. Lipase-catalyzed acidolysis of palm mid fraction oil with palmitic and stearic Fatty Acid mixture for production of cocoa butter equivalent.

    PubMed

    Mohamed, Ibrahim O

    2013-10-01

    Cocoa butter equivalent (CBE) was prepared by enzymatic acidolysis reaction of substrate consisting of refined palm mid fraction oil and palmitic-stearic fatty acid mixture. The reactions were performed in a batch reactor at a temperature of 60 °C in an orbital shaker operated at 160 RPM. Different mass ratios of substrates were explored, and the composition of the five major triacylglycerols (TAGs) of the structured lipids was identified and quantified using cocoa butter certified reference material IRMM-801. The reaction resulted in production of cocoa butter equivalent with the TAGs' composition (1,3-dipalmitoyl-2-oleoyl-glycerol 30.7%, 1-palmitoyl-2-oleoyl-3-stearoyl-rac-glycerol 40.1%, 1-palmitoy-2,3- dioleoyl glycerol 9.0%, 1,3-distearoyl-2-oleoyl-glycerol 14.5 %, and 1-stearoyl-2,3-dioleoyl glycerol 5.7%) and with onset melting temperature of 31.6 °C and peak temperature of 40.4 °C which are close to those of cocoa butter. The proposed kinetics model for the acidolysis reaction presented the experimental data very well. The results of this research showed that palm mid fraction oil TAGs could be restructured to produce value added product such as CBE.

  10. Characterization of the O-antigen polymerase (Wzy) of Francisella tularensis.

    PubMed

    Kim, Tae-Hyun; Sebastian, Shite; Pinkham, Jessica T; Ross, Robin A; Blalock, LeeAnn T; Kasper, Dennis L

    2010-09-03

    The O-antigen polymerase of gram-negative bacteria has been difficult to characterize. Herein we report the biochemical and functional characterization of the protein product (Wzy) of the gene annotated as the putative O-antigen polymerase, which is located in the O-antigen biosynthetic locus of Francisella tularensis. In silico analysis (homology searching, hydropathy plotting, and codon usage assessment) strongly suggested that Wzy is an O-antigen polymerase whose function is to catalyze the addition of newly synthesized O-antigen repeating units to a glycolipid consisting of lipid A, inner core polysaccharide, and one repeating unit of the O-polysaccharide (O-PS). To characterize the function of the Wzy protein, a non-polar deletion mutant of wzy was generated by allelic replacement, and the banding pattern of O-PS was observed by immunoblot analysis of whole-cell lysates obtained by SDS-PAGE and stained with an O-PS-specific monoclonal antibody. These immunoblot analyses showed that O-PS of the wzy mutant expresses only one repeating unit of O-antigen. Further biochemical characterization of the subcellular fractions of the wzy mutant demonstrated that (as is characteristic of O-antigen polymerase mutants) the low molecular weight O-antigen accumulates in the periplasm of the mutant. Site-directed mutagenesis based on protein homology and topology, which was carried out to locate a catalytic residue of the protein, showed that modification of specific residues (Gly(176), Asp(177), Gly(323), and Tyr(324)) leads to a loss of O-PS polymerization. Topology models indicate that these amino acids most likely lie in close proximity on the bacterial surface.

  11. Characterization of the O-antigen Polymerase (Wzy) of Francisella tularensis*

    PubMed Central

    Kim, Tae-Hyun; Sebastian, Shite; Pinkham, Jessica T.; Ross, Robin A.; Blalock, LeeAnn T.; Kasper, Dennis L.

    2010-01-01

    The O-antigen polymerase of Gram-negative bacteria has been difficult to characterize. Herein we report the biochemical and functional characterization of the protein product (Wzy) of the gene annotated as the putative O-antigen polymerase, which is located in the O-antigen biosynthetic locus of Francisella tularensis. In silico analysis (homology searching, hydropathy plotting, and codon usage assessment) strongly suggested that Wzy is an O-antigen polymerase whose function is to catalyze the addition of newly synthesized O-antigen repeating units to a glycolipid consisting of lipid A, inner core polysaccharide, and one repeating unit of the O-polysaccharide (O-PS). To characterize the function of the Wzy protein, a non-polar deletion mutant of wzy was generated by allelic replacement, and the banding pattern of O-PS was observed by immunoblot analysis of whole-cell lysates obtained by SDS-PAGE and stained with an O-PS-specific monoclonal antibody. These immunoblot analyses showed that O-PS of the wzy mutant expresses only one repeating unit of O-antigen. Further biochemical characterization of the subcellular fractions of the wzy mutant demonstrated that (as is characteristic of O-antigen polymerase mutants) the low molecular weight O-antigen accumulates in the periplasm of the mutant. Site-directed mutagenesis based on protein homology and topology, which was carried out to locate a catalytic residue of the protein, showed that modification of specific residues (Gly176, Asp177, Gly323, and Tyr324) leads to a loss of O-PS polymerization. Topology models indicate that these amino acids most likely lie in close proximity on the bacterial surface. PMID:20605777

  12. Gallic Acid Enriched Fraction of Phyllanthus emblica Potentiates Indomethacin-Induced Gastric Ulcer Healing via e-NOS-Dependent Pathway.

    PubMed

    Chatterjee, Ananya; Chatterjee, Sirshendu; Biswas, Angshuman; Bhattacharya, Sayanti; Chattopadhyay, Subrata; Bandyopadhyay, Sandip K

    2012-01-01

    The healing activity of gallic acid enriched ethanolic extract (GAE) of Phyllanthus emblica fruits (amla) against the indomethacin-induced gastric ulceration in mice was investigated. The activity was correlated with the ability of GAE to alter the cyclooxygenase- (COX-) dependent healing pathways. Histology of the stomach tissues revealed maximum ulceration on the 3rd day after indomethacin (18 mg/kg, single dose) administration that was associated with significant increase in inflammatory factors, namely, mucosal myeloperoxidase (MPO) activity and inducible nitric oxide synthase (i-NOS) expression. Proangiogenic parameters such as the levels of prostaglandin (PG) E(2), vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), von Willebrand Factor VIII, and endothelial NOS (e-NOS) were downregulated by indomethacin. Treatment with GAE (5 mg/kg/day) and omeprazole (3 mg/kg/day) for 3 days led to effective healing of the acute ulceration, while GAE could reverse the indomethacin-induced proinflammatory changes of the designated biochemical parameters. The ulcer healing activity of GAE was, however, compromised by coadministration of the nonspecific NOS inhibitor, N-nitro-L-arginine methyl ester (L-NAME), but not the i-NOS-specific inhibitor, L-N6-(1-iminoethyl) lysine hydrochloride (L-NIL). Taken together, these results suggested that the GAE treatment accelerates ulcer healing by inducing PGE(2) synthesis and augmenting e-NOS/i-NOS ratio.

  13. To assess of behavior of natural colloid (soil extraction and fractionation of natural water) humus acid in comparison with synthetic humus acid

    NASA Astrophysics Data System (ADS)

    Dinu, Marina; Shkinev, Valery

    2015-04-01

    To study and predict the fate of natural colloid - nanoparticles in surface water or soil extraction - necessary to understand the features of the migration and physic-chemical activity of biocolloids. Comparison of the behavior of natural biocolloids, such as humus acid extracts of soil or natural water with artificial, synthetic humic acids (introduced into the environment) allows you to explore the mechanism of formation and transformation biocolloids under the influence of a number of parameters. In this work, we studied these interactions in natural surface waters from lakes and soil (Russian Federation, Kola North and Western Siberia) which displaying contrast organic and inorganic compositions. During the study, researches identified zonal features influence on the qualitative and quantitative composition of colloids, their stability and chemical activity. A model approach was also followed with synthetic water of comparable composition in order to better understand the driving mechanisms. We investigated the size, zeta potential and other physical and chemical parameters of the system. Particular attention is given to the process of complexation with heavy metal ions. As humic substances have excellent complexion properties and reduce the toxicity of many metal ions. The study of such non-static natural systems allow studying the features of the existence of natural colloidal components. The use of synthetic humic substances, which were introduced into the natural environment possible to study the standard mechanisms of formation, development and destruction of colloidal polymer systems. The obtained results allowed with used computer programs MatnLab, MathCad, Statistics simulate the processes of formation, development and functioning of natural colloids.

  14. Profile of Free Fatty Acids and Fractions of Phospholipids, Cholesterol Esters and Triglycerides in Serum of Obese Youth with and without Metabolic Syndrome

    PubMed Central

    Bermúdez-Cardona, Juliana; Velásquez-Rodríguez, Claudia

    2016-01-01

    The study evaluated the profile of circulating fatty acids (FA) in obese youth with and without metabolic syndrome (MetS) to determine its association with nutritional status, lifestyle and metabolic variables. A cross-sectional study was conducted in 96 young people, divided into three groups: obese with MetS (OBMS), obese (OB) and appropriate weight (AW). FA profiles were quantified by gas chromatography; waist circumference (WC), fat folds, lipid profile, high-sensitivity C-reactive protein, glucose, insulin, the homeostasis model assessment (HOMA index), food intake and physical activity (PA) were assessed. The OBMS group had significantly greater total free fatty acids (FFAs), palmitic-16:0 in triglyceride (TG), palmitoleic-16:1n-7 in TG and phospholipid (PL); in the OB group, these FAs were higher than in the AW group. Dihomo-gamma-linolenic (DHGL-20:3n-6) was higher in the OBMS than the AW in PL and FFAs. Linoleic-18:2n-6 in TG and PL had the lowest proportion in the OBMS group. WC, PA, total FFA, linoleic-18:2n-6 in TG and DHGL-20:3n-6 in FFAs explained 62% of the HOMA value. The OB group presented some higher proportions of FA and biochemical values than the AW group. The OBMS had proportions of some FA in the TG, PL and FFA fractions that correlated with disturbances of MetS. PMID:26891317

  15. Evaluation of antifungal activity of free fatty acids methyl esters fraction isolated from Algerian Linum usitatissimum L. seeds against toxigenic Aspergillus

    PubMed Central

    Abdelillah, Amrouche; Houcine, Benmehdi; Halima, Dalile; Meriem, Chabane sari; Imane, Zaaboub; Eddine, Smahi Djamal; Abdallah, Moussaoui; Daoudi, Chabane sari

    2013-01-01

    Objective The aim of this study was to evaluate the antifungal activity of the major fraction of fatty acids methyl esters (FAMEs) isolated from Linum usitatissimum L. seeds oil collected from Bechar department (Algeria). Methods The assessment of antifungal activity was carried out in terms of percentage of radial growth on solid medium (potatoes dextrose agar PDA) and biomass growth inhibition on liquid medium (potatoes dextrose broth PDB) against two fungi. Results The FAMEs was found to be effective in inhibiting the radial mycelial growth of Aspergillus flavus more than Aspergillus ochraceus on all tested concentrations. The highest antifungal index was found to be (54.19%) compared to Aspergillus ochraceus (40.48%). The results of the antifungal activity of the FAMEs inhibition of biomass on liquid medium gave no discounted results, but this does not exclude the antifungal activity. Conclusions We can assume that the observed antifungal potency may be due to the abundance of linoleic and α-linolenic acids in linseed oil which appears to be promising to treat fungal infections, storage fungi and food spoilage in food industry field. PMID:23730556

  16. Calcium-dependent antigen binding as a novel modality for antibody recycling by endosomal antigen dissociation.

    PubMed

    Hironiwa, N; Ishii, S; Kadono, S; Iwayanagi, Y; Mimoto, F; Habu, K; Igawa, T; Hattori, K

    2016-01-01

    The pH-dependent antigen binding antibody, termed a recycling antibody, has recently been reported as an attractive type of second-generation engineered therapeutic antibody. A recycling antibody can dissociate antigen in the acidic endosome, and thus bind to its antigen multiple times. As a consequence, a recycling antibody can neutralize large amounts of antigen in plasma. Because this approach relies on histidine residues to achieve pH-dependent antigen binding, which could limit the epitopes that can be targeted and affect the rate of antigen dissociation in the endosome, we explored an alternative approach for generating recycling antibodies. Since calcium ion concentration is known to be lower in endosome than in plasma, we hypothesized that an antibody with antigen-binding properties that are calcium-dependent could be used as recycling antibody. Here, we report a novel anti-interleukin-6 receptor (IL-6R) antibody, identified from a phage library that binds to IL-6R only in the presence of a calcium ion. Thermal dynamics and a crystal structure study revealed that the calcium ion binds to the heavy chain CDR3 region (HCDR3), which changes and possibly stabilizes the structure of HCDR3 to make it bind to antigen calcium dependently (PDB 5AZE). In vitro and in vivo studies confirmed that this calcium-dependent antigen-binding antibody can dissociate its antigen in the endosome and accelerate antigen clearance from plasma, making it a novel approach for generating recycling antibody.

  17. Characterization of antigenic components from circulating immune complexes in patients with gestational trophoblastic neoplasia.

    PubMed

    Lahey, S J; Steele, G; Rodrick, M L; Berkowitz, R; Goldstein, D P; Ross, D S; Ravikumar, T S; Wilson, R E; Byrn, R; Thomas, P

    1984-03-15

    The authors have studied serial circulating immune complex (CIC) levels in 15 patients with gestational trophoblastic neoplasia (GTN) for several reasons. Gestational trophoblastic neoplasia can easily be followed from presentation to remission, and CIC changes can be compared with changes in human chorionic gonadotropin (HCG) which is a specific and quantitative marker of trophoblastic tumor load. Twelve patients with hydatidiform molar pregnancy presented with normal CIC levels (255 delta OD450 +/- 97, mean SE [standard error]) as measured by our antigen nonspecific polyethylene glycol (PEG) turbidity assay. Only after reduction in tumor load as monitored by a fall in HCG did CIC rise. In contrast, three patients with choriocarcinoma presented with significantly elevated CIC levels (513 delta OD450 +/- 147, P less than 0.05 compared to normals) which slowly declined in parallel with HCG levels following evacuation and chemotherapy. Sera at peak PEG-CIC from three patients with molar pregnancy or choriocarcinoma were precipitated with 3.75% polyethylene glycol to concentrate circulating immune complexes. Circulating immune complex levels were fractionated on Sephadex G-200 in an acid buffer (pH = 2.8). An identifiable antigenic component of the CIC in both diseases was found to be paternal HLA antigen. This was demonstrated by the ability of the latest eluting CIC fraction to inhibit paternal lymphocyte lysis using anti-HLA antisera against the husband's HLA tissue type. In each case, this fraction contained no immunoglobulin or beta-2 microglobulin and was antigenically crossreactive with only one of the husband's HLA haplotypes. The authors believe the PEG-CIC assay has allowed them to define the kinetics of host humoral response in GTN, and has provided a method for recovering immunogenic tumor-associated antigens from these complexes which may apply to other solid tumors.

  18. Cyclohexane-1,2-dicarboxylic acid diisononyl ester and metabolite effects on rat epididymal stromal vascular fraction differentiation of adipose tissue

    SciTech Connect

    Campioli, Enrico; Duong, Tam B.; Deschamps, François; Papadopoulos, Vassilios

    2015-07-15

    Plastics are generally mixed with additives like plasticizers to enhance their flexibility, pliability, and elasticity proprieties. Plasticizers are easily released into the environment and are absorbed mainly through ingestion, dermal contact, and inhalation. One of the main classes of plasticizers, phthalates, has been associated with endocrine and reproductive diseases. In 2002, 1,2-cyclohexane dicarboxylic acid diisononyl ester (DINCH) was introduced in the market for use in plastic materials and articles intended to come into contact with food, and it received final approval from the European Food Safety Authority in 2006. At present, there is limited knowledge about the safety and potential metabolic and endocrine-disrupting properties of DINCH and its metabolites. The purpose of this study was to evaluate the biological effects of DINCH and its active metabolites, cyclohexane-1,2-dicarboxylic acid (CHDA) and cyclohexane-1,2-dicarboxylic acid mono isononyl ester (MINCH), on rat primary stromal vascular fraction (SVF) of adipose tissue. DINCH and its metabolite, CHDA, were not able to directly affect SVF differentiation. However, exposure of SVF to 50 μM and 100 μM concentrations of MINCH affected the expression of Cebpa and Fabp4, thus inducing SVF preadipocytes to accumulate lipids and fully differentiate into mature adipocytes. The effect of MINCH was blocked by the specific peroxisome proliferator-activated receptor (PPAR)-α antagonist, GW6471. Taken together, these results suggest that MINCH is a potent PPAR-α agonist and a metabolic disruptor, capable of inducing SVF preadipocyte differentiation, that may interfere with the endocrine system in mammals. - Highlights: • DINCH and CHDA did not affect the adipogenesis of the SVF. • MINCH affected the adipogenesis of the SVF. • MINCH effect was blocked by the specific PPAR-α antagonist GW6471. • MINCH exerted a similar effect as MEHP on SVF adipogenesis. • DINCH/MINCH are potential metabolic

  19. Immune responses to vaccines involving a combined antigen-nanoparticle mixture and nanoparticle-encapsulated antigen formulation.

    PubMed

    Zhang, Weifeng; Wang, Lianyan; Liu, Yuan; Chen, Xiaoming; Liu, Qi; Jia, Jilei; Yang, Tingyuan; Qiu, Shaohui; Ma, Guanghui

    2014-07-01

    Many physicochemical characteristics significantly influence the adjuvant effect of micro/nanoparticles; one critical factor is the kinetics of antigen exposure to the immune system by particle-adjuvanted vaccines. Here, we investigated how various antigen-nanoparticle formulations impacted antigen exposure to the immune system and the resultant antigen-specific immune responses. We formulated antigen with poly(lactic-co-glycolic acid) (PLGA) nanoparticles by encapsulating antigen within nanoparticles or by simply mixing soluble antigen with the nanoparticles. Our results indicated that the combined formulation (composed of antigen encapsulated in nanoparticles and antigen mixed with nanoparticles) induced more powerful antigen-specific immune responses than each single-component formulation. Mice immunized with the combined vaccine formulation displayed enhanced induction of antigen-specific IgG antibodies with high avidity, increased cytokine secretion by splenocytes, and improved generation of memory T cell. Enhanced immune responses elicited by the combined vaccine formulation might be attributed to the antigen-depot effect at the injection site, effective provision of both adequate initial antigen exposure and long-term antigen persistence, and efficient induction of dendritic cell (DC) activation and follicular helper T cell differentiation in draining lymph nodes. Understanding the effect of antigen-nanoparticle formulations on the resultant immune responses might have significant implications for rational vaccine design.

  20. Use of HLA peptidomics and whole exome sequencing to identify human immunogenic neo-antigens

    PubMed Central

    Kalaora, Shelly; Qutob, Nouar; Teer, Jamie K.; Shimony, Nilly; Schachter, Jacob; Rosenberg, Steven A.; Samuels, Yardena

    2016-01-01

    The antigenicity of cells is demarcated by the peptides bound by their Human Leucocyte Antigen (HLA) molecules. Through this antigen presentation, T cell specificity response is controlled. As a fraction of the expressed mutated peptides is presented on the HLA, these neo-epitopes could be immunogenic. Such neoantigens have recently been identified through screening for predicted mutated peptides, using synthetic peptides or ones expressed from minigenes, combined with screening of patient tumor-infiltrating lymphocytes (TILs). Here we present a time and cost-effective method that combines whole-exome sequencing analysis with HLA peptidome mass spectrometry, to identify neo-antigens in a melanoma patient. Of the 1,019 amino acid changes identified through exome sequencing, two were confirmed by mass spectrometry to be presented by the cells. We then synthesized peptides and evaluated the two mutated neo-antigens for reactivity with autologous bulk TILs, and found that one yielded mutant-specific T-cell response. Our results demonstrate that this method can be used for immune response prediction and promise to provide an alternative approach for identifying immunogenic neo-epitopes in cancer. PMID:26819371

  1. Absorption of inhaled antigen into the circulation of isolated lungs from normal and immunized rabbits.

    PubMed Central

    Braley, J F; Dawson, C A; Moore, V L; Cozzini, B O

    1978-01-01

    The purpose of this study was to determine whether the absorption of inhaled antigen (Ag) across the pulmonary air-blood barrier of the isolated perfused lung can be modulated by immunologic mechanisms. Lungs from immunized or nonimmunized rabbits were removed, ventilated, and perfused with autochthonous blood. Radioiodinated Ag (human serum albumin or ovalbumin) was introduced as an aerosol into the isolated lung for 15 min and blood samples were taken over a 4-h period. The results showed that radioactivity fom inhaled Ag entered the perfusing blood as two fractions. One fraction was precipitable by 5% trichloroacetic acid or antiserum. The TCA-soluble fraction chromatographed differently from iodide and may have represented metabolites of the Ag. Immunization specifically reduced the amount of antigenically intact protein entering the blood. On the other hand, the metabolite reached higher concentrations in the blood of immunized lungs. We conclude that the alveolar capillary barrier of the normal rabbit lung could provide a significant route of entry for inhaled antigen into the systemic circulation and that immunization reduces absorption via this route and enhances pulmonary metabolism of the Ag. PMID:678344

  2. Fractionation and identification of 9c, 11t, 13t-conjugated linolenic acid as an activator of PPARalpha in bitter gourd (Momordica charantia L.).

    PubMed

    Chuang, Chia-Ying; Hsu, Chin; Chao, Che-Yi; Wein, Yung-Shung; Kuo, Yueh-Hsiung; Huang, Ching-jang

    2006-11-01

    Bitter gourd (Momordica charantia L.) is a common vegetable in Asia that has been used in traditional medicine for the treatment of Diabetes. PPARs are ligand-dependent transcription factors that belong to the steroid hormone nuclear receptor family and control lipid and glucose homeostasis in the body. We previously reported that the ethyl acetate (EA) extract of bitter gourd activated peroxisome proliferator receptors (PPARs) alpha and gamma. To identify the active compound that activated PPARalpha, wild bitter gourd EA extract was partitioned between n-hexane and 90% methanol/10% H(2)O, and the n-hexane soluble fraction was further separated by silica gel column chromatography and finally by preparative HPLC. A transactivation assay employing a clone of CHOK1 cells stably transfected with a (UAS)(4)-tk-alkaline phosphatase reporter and a chimeric receptor of GAL4-rPPARalpha LBD was used to track the active component. Based on Mass, NMR, and IR spectroscopy, 9cis, 11trans, 13trans-conjugated linolenic acid (9c, 11t, 13t-CLN) was identified as a PPARalpha activator in wild bitter gourd. The isolated 9c, 11t, 13t-CLN rich fraction also significantly induced acyl CoA oxidase (ACO) activity in a peroxisome proliferator-responsive murine hepatoma cell line, H4IIEC3, implying that 9c, 11t, 13t-CLN was able to act on a natural PPARalpha signaling pathway as well. The content of 9c, 11t, 13t-CLN was estimated to be about 7.1 g/kg of our dried wild bitter gourd sample. The concentration of 9c, 11t, 13t-CLN and activation activity in the hydrolyzed EA extract of the seeds was higher than that of the flesh. The potential health benefits of 9c, 11t, 13t-CLN through the PPARalpha regulated mechanism are worthy to be further characterized in in vivo studies.

  3. Detection of functional class II-associated antigen: role of a low density endosomal compartment in antigen processing

    PubMed Central

    1995-01-01

    We have developed a functional assay to identify processed antigen in subcellular fractions from antigen-presenting cells; stimulatory activity in this assay may be caused by either free peptide fragments or by complexes of peptide fragments and class II molecules present on organellar membrane sheets and vesicles. In addition, we have developed a functional assay to identify proteolytic activity in subcellular fractions capable of generating antigenic peptides from intact proteins. These techniques permit the direct identification of intracellular sites of antigen processing and class II association. Using a murine B cell line stably transfected with a phosphorylcholine (PC)-specific membrane-bound immunoglobulin (Ig), we show that PC- conjugated antigens are rapidly internalized and efficiently degraded to generate processed antigen within an early low density compartment. Proteolytic activity capable of generating antigenic peptide fragments from intact proteins is found within low density endosomes and a dense compartment consistent with lysosomes. However, neither processed peptide nor peptide-class II complexes are detected in lysosomes from antigen-pulsed cells. Furthermore, blocking the intracellular transport of internalized antigen from the low density endosome to lysosomes does not inhibit the generation of processed antigen. Therefore, antigens internalized in association with membrane Ig on B cells can be efficiently processed in low density endosomal compartments without the contribution of proteases present within denser organelles. PMID:7722450

  4. Genetic and antigenic changes in porcine rubulavirus.

    PubMed

    Sánchez-Betancourt, José I; Trujillo, María E; Mendoza, Susana E; Reyes-Leyva, Julio; Alonso, Rogelio A

    2012-01-01

    Blue eye disease, caused by a porcine rubulavirus (PoRV), is an emergent viral swine disease that has been endemic in Mexico since 1980. Atypical outbreaks were detected in 1990 and 2003. Growing and adult pigs presented neurological signs, mild neurological signs were observed in piglets, and severe reproductive problems were observed in adults. Amino acid sequence comparisons and phylogenetic analysis of the hemagglutinin-neuraminidase (HN) protein revealed genetically different lineages. We used cross-neutralization assays, with homologous and heterologous antisera, to determine the antigenic relatedness values for the PoRV isolates. We found antigenic changes among several strains and identified a highly divergent one, making up a new serogroup. It seems that genetically and antigenically different PoRV strains are circulating simultaneously in the swine population in the geographical region studied. The cross neutralization studies suggest that the HN is not the only antigenic determinant participating in the antigenic changes among the different PoRV strains.

  5. A Substantial Fraction of Barley (Hordeum vulgare L.) Low Phytic Acid Mutations Have Little or No Effect on Yield across Diverse Production Environments.

    PubMed

    Raboy, Victor; Peterson, Kevin; Jackson, Chad; Marshall, Juliet M; Hu, Gongshe; Saneoka, Hirofumi; Bregitzer, Phil

    2015-04-29

    The potential benefits of the low phytic acid (lpa) seed trait for human and animal nutrition, and for phosphorus management in non-ruminant animal production, are well documented. However, in many cases the lpa trait is associated with impaired seed or plant performance, resulting in reduced yield. This has given rise to the perception that the lpa trait is tightly correlated with reduced yield in diverse crop species. Here we report a powerful test of this correlation. We measured grain yield in lines homozygous for each of six barley (Hordeum vulgare L.) lpa mutations that greatly differ in their seed phytic acid levels. Performance comparisons were between sibling wild-type and mutant lines obtained following backcrossing, and across two years in five Idaho (USA) locations that greatly differ in crop yield potential. We found that one lpa mutation (Hvlpa1-1) had no detectable effect on yield and a second (Hvlpa4-1) resulted in yield losses of only 3.5%, across all locations. When comparing yields in three relatively non-stressful production environments, at least three lpa mutations (Hvlpa1-1, Hvlpa3-1, and Hvlpa4-1) typically had yields similar to or within 5% of the wild-type sibling isoline. Therefore in the case of barley, lpa mutations can be readily identified that when simply incorporated into a cultivar result in adequately performing lines, even with no additional breeding for performance within the lpa line. In conclusion, while some barley lpa mutations do impact field performance, a substantial fraction appears to have little or no effect on yield.

  6. Understanding Multiplication of Fractions.

    ERIC Educational Resources Information Center

    Sweetland, Robert D.

    1984-01-01

    Discussed the use of Cuisenaire rods in teaching the multiplication of fractions. Considers whole number times proper fraction, proper fraction multiplied by proper fraction, mixed number times proper fraction, and mixed fraction multiplied by mixed fractions. (JN)

  7. A study on the use of caprylic acid and ammonium sulfate in combination for the fractionation of equine antivenom F(ab')(2).

    PubMed

    Simsiriwong, Pavinee; Eursakun, Sukanya; Ratanabanangkoon, Kavi

    2012-09-01

    This study involved the fractionation of equine antivenom F(ab')(2) by combined stepwise ammonium sulfate (AS) and caprylic acid (CA) precipitation without intermediate separation of precipitate. Using a microplate format, 55 conditions with combinations of AS (0-20% saturation) and CA (0-5% v/v), were tested. AS significantly reduced the turbidity raised by CA. High specific antibody activity was observed in the area containing 2-5% CA and 10-20% AS. From these results, 12 precipitation conditions were selected for detailed quantitative studies. Two combinations, one with 4% CA and 15% AS and another with 5% CA and 20% AS, gave the highest fold-purification (1.79 and 1.83) with antibody recoveries at 68% and 59%, respectively. These combinations offered a benefit over CA alone in reducing the turbidity and in increasing the purity but not the recovery of antibody. The conditions giving more favorable overall results were with 2% CA alone and another with a combination of 1.5% CA and 10% AS. These preparations of F(ab')(2) were homogeneous and without protein aggregate under size-exclusion HPLC. Lastly, 1 h precipitation showed better results than those of overnight precipitation. These results could be useful for the production of therapeutic antivenoms.

  8. Split-face comparative study of 1550 nm fractional photothermolysis and trichloroacetic acid 15% chemical peeling for facial melasma in Asian skin.

    PubMed

    Hong, Seung-Phil; Han, Seung-Seog; Choi, Seok-Joo; Kim, Myoung-Shin; Won, Chong-Hyun; Lee, Mi-Woo; Choi, Jee-Ho; Moon, Kee-Chan; Kim, Youn Jin; Chang, Sung-Eun

    2012-04-01

    Fractional photothermolysis (FP) therapy and chemical peels have been reported to be effective in patients with recalcitrant melasma. However, there is little information to compare the efficacy of single treatment session in Asian women. The aim of this study was to examine the efficacy, long-lasting outcomes and safety of a single session of 1550-nm erbium-doped FP in Asian patients, compared with trichloroacetic acid (TCA) peel with a medium depth. Eighteen Korean women (Fitzpatrick skin type III or IV) with moderate-to-severe bilateral melasma were randomly treated with a single session of 1550-nm FP on one cheek, and with a 15% TCA peel on the other cheek. Outcome measures included an objective melasma area severity index and subjective patient-rated overall improvement at 4 and 12 weeks after treatment. Melasma lesions were significantly improved 4 weeks after either treatment, but melasma recurred at 12 weeks. Post-inflammatory hyperpigmentation developed in 28% of patients at 4 weeks but resolved in all but one patient by 12 weeks. There was no difference between FP treatment and TCA peeling with respect to any outcome measure. FP laser and TCA peel treatments were equally effective and safe when used to treat moderate-to-severe melasma, but neither treatment was long-lasting. We suggest that multiple or periodic maintenance treatments and/or supplemental procedures may be required for the successful treatment of melasma in Asian women.

  9. Diagnostic performances of antigen detection compared to conventional and nucleic acid detection of Entamoeba histolytica in a non-endemic setting.

    PubMed

    Calderaro, Adriana; Piergianni, Maddalena; Piccolo, Giovanna; Rossi, Sabina; Montecchini, Sara; Buttrini, Mirko; Arcangeletti, Maria Cristina; Medici, Maria Cristina; Chezzi, Carlo; De Conto, Flora

    2016-04-01

    This study evaluated the immunochromatographic (IC) assay "TECHLAB(®) E. HISTOLYTICA QUIK CHEK™" analysing 36 faecal samples and 7 cultured strains. This assay was compared to the methods performed in our laboratory for the diagnosis of amoebiasis. The IC assay revealed a detection limit of 103 trophozoites/g faeces and no cross-reactivity with other parasites and failed to detect E. histolytica antigen in frozen faeces. In our laboratory located in a non-endemic setting this assay could not replace the methods currently used for the diagnosis of amoebiasis.

  10. Mystery Fractions

    ERIC Educational Resources Information Center

    Bhattacharyya, Sonalee; Namakshi, Nama; Zunker, Christina; Warshauer, Hiroko K.; Warshauer, Max

    2016-01-01

    Making math more engaging for students is a challenge that every teacher faces on a daily basis. These authors write that they are constantly searching for rich problem-solving tasks that cover the necessary content, develop critical-thinking skills, and engage student interest. The Mystery Fraction activity provided here focuses on a key number…

  11. Pitch Fractionation.

    DTIC Science & Technology

    1981-12-15

    13 3. Solvent Fractionation Experiments .................................... 15 4. Fourier Transform Infrared Spectra for A240 Petrolem Pitch AG 12...34 and Mesophase Pitch AG 164B ............................... 21 5. Fourier Transform Infrared Spectra ................................... 23 6...compared by Fourier transform infrared (FTIR) analysis using a Digilab Model FTS 14 spectrophotometer (Rockwell International, Anaheim, California

  12. Proteomic analysis of different period excretory secretory products from Clonorchis sinensis adult worms: molecular characterization, immunolocalization, and serological reactivity of two excretory secretory antigens-methionine aminopeptidase 2 and acid phosphatase.

    PubMed

    Zheng, Minghui; Hu, Kunhua; Liu, Wei; Li, Hongyu; Chen, Jingfang; Yu, Xinbing

    2013-03-01

    The excretory secretory products (ESP) of Clonorchis sinensis are the causative agents of clonorchiasis and biliary diseases. The parasites' ESP play important roles in host-parasite interactions. The protein compositions of ESP at different secretory times are different and have not been systemically investigated so far. In this study, we collected ESP from six different periods (0-3 h, 3-6 h, 6-12 h, 12-24 h, 24-36 h, and 36-48 h) from C. sinensis adults. Using a shotgun LC-MS/MS analysis, we found 187, 80, 103, 58, 248, and 383 proteins, respectively. Among these proteins, we selected methionine aminopeptidase 2 (MAP-2, presented in 24-36 h and 36-48 h ESP) and acid phosphatase (AP, presented in 3-6 h, 12-24 h, 24-36 h, and 36-48 h ESP) for further study. Bioinformatics analysis showed that CsMAP-2 has metallopeptidase family M24, unique lysine residue-rich and acidic residue-rich domain, SGTS motif, and auto-cleavage point; and that CsAP has possible signal sequence cleavage site, acid phosphate domain, and two histidine acid phosphatases active regions. CsMAP-2 and CsAP's cDNA have 1,425 bp and1,410 bp ORF, encoding 475 and 470 amino acid proteins and weighing 55.3840 kDa and 55.2875 kDa, respectively. MAP-2 and AP were identified as antigens present in the ESP and circulating antigens by immunoblot analysis, which were also found expressing in the eggs, metacercaria, and adult stages of C. sinensis. Immunofluorescence analysis showed that they were located in tegument and intestinal cecum of adult. MTT assay showed that they could inhibit hepatic stellate cell line (LX-2) proliferation. These findings presented the compositions of different period excretory secretary products from C. sinensis adults.

  13. Adsorption and fractionation of a muck fulvic acid on kaolinite and goethite at pH 3.7,6, and 8

    USGS Publications Warehouse

    Namjesnik-Dejanovic, K.; Maurice, P.A.; Aiken, G.R.; Cabaniss, S.; Chin, Y.-P.; Pullin, M.J.

    2000-01-01

    Molecular weight (MW) of humic materials is a key factor controlling proton and metal binding and organic pollutant partitioning. Several studies have suggested preferential adsorption of higher MW, more aromatic moieties to mineral surfaces; quantification of such processes is fundamental to development of predictive models. We used high pressure size exclusion chromatography (HPSEC) to quantify MW changes upon adsorption of a muck fulvic acid (MFA) extracted from a peat deposit to kaolinite and goethite, at pH 3.7,6, and 8 at 22 ??C, I = 0.01 (NaC1), 24-h reaction time. MFA adsorption affinity was greater for goethite than for kaolinite. At concentrations less than the adsorption maximum (A(max)) for both adsorbents, the weight-average MW (M(w)) of MFA remaining in solution decreased by as much as several hundred Daltons relative to control samples, indicating preferential adsorption of the higher MW components. At concentrations more than A(max), M(w) of MFA in solution did not change appreciably. Although total adsorption decreased significantly as pH increased, fractionation as measured by change in M(w) remained similar, perhaps indicating greater selectivity for higher MW components at higher pH. Absorptivities at ?? = 280 nm normalized to mg C L-1 (??) suggested preferential adsorption of more aromatic moieties to kaolinite. ?? could not be used for goethite-reacted samples because high Fe concentrations in the aqueous phase brought about by goethite dissolution interfered with the spectroscopic analysis. Preliminary kinetic experiments suggested that smaller molecules adsorbed first and were replaced by larger molecules whose adsorption was thermodynamically favored.

  14. Levels and Age Dependency of Neurofilament Light and Glial Fibrillary Acidic Protein in Healthy Individuals and Their Relation to the Brain Parenchymal Fraction

    PubMed Central

    Vågberg, Mattias; Norgren, Niklas; Dring, Ann; Lindqvist, Thomas; Birgander, Richard; Zetterberg, Henrik; Svenningsson, Anders

    2015-01-01

    Background Neurofilament light (NFL) and Glial Fibrillary Acidic Protein (GFAP) are integral parts of the axonal and astrocytal cytoskeletons respectively and are released into the cerebrospinal fluid (CSF) in cases of cellular damage. In order to interpret the levels of these biomarkers in disease states, knowledge on normal levels in the healthy is required. Another biomarker for neurodegeneration is brain atrophy, commonly measured as brain parenchymal fraction (BPF) using magnetic resonance imaging (MRI). Potential correlations between levels of NFL, GFAP and BPF in healthy individuals have not been investigated. Objectives To present levels of NFL and GFAP in healthy individuals stratified for age, and investigate the correlation between them as well as their correlation with BPF. Methods The CSF was analysed in 53 healthy volunteers aged 21 to 70 (1 sample missing for GFAP analysis) and 48 of the volunteers underwent determination of BPF using MRI. Results Mean (±SD) NFL was 355 ng/L (±214), mean GFAP was 421 ng/L (±129) and mean BPF was 0.867 (±0.035). All three biomarkers correlated with age. NFL also correlated with both GFAP and BPF. When controlled for age, only the correlation between NFL and GFAP retained statistical significance. Conclusions This study presents data on age-stratified levels of NFL and GFAP in the CSF of healthy individuals. There is a correlation between levels of NFL and GFAP and both increase with age. A correlation between NFL and BPF was also found, but did not retain statistical significance if controlled for age. PMID:26317831

  15. Comparative Study of Corn Stover Pretreated by Dilute Acid and Cellulose Solvent-Based Lignocellulose Fractionation: Enzymatic Hydrolysis, Supramolecular Structure, and Substrate Accessibility

    SciTech Connect

    Zhu, Z.; Sathitsuksanoh, N.; Vinzant, T.; Schell, D. J.; McMillian, J. D.; Zhang, Y. H. P.

    2009-07-01

    Liberation of fermentable sugars from recalcitrant biomass is among the most costly steps for emerging cellulosic ethanol production. Here we compared two pretreatment methods (dilute acid, DA, and cellulose solvent and organic solvent lignocellulose fractionation, COSLIF) for corn stover. At a high cellulase loading [15 filter paper units (FPUs) or 12.3 mg cellulase per gram of glucan], glucan digestibilities of the corn stover pretreated by DA and COSLIF were 84% at hour 72 and 97% at hour 24, respectively. At a low cellulase loading (5 FPUs per gram of glucan), digestibility remained as high as 93% at hour 24 for the COSLIF-pretreated corn stover but reached only {approx}60% for the DA-pretreated biomass. Quantitative determinations of total substrate accessibility to cellulase (TSAC), cellulose accessibility to cellulase (CAC), and non-cellulose accessibility to cellulase (NCAC) based on adsorption of a non-hydrolytic recombinant protein TGC were measured for the first time. The COSLIF-pretreated corn stover had a CAC of 11.57 m{sup 2}/g, nearly twice that of the DA-pretreated biomass (5.89 m{sup 2}/g). These results, along with scanning electron microscopy images showing dramatic structural differences between the DA- and COSLIF-pretreated samples, suggest that COSLIF treatment disrupts microfibrillar structures within biomass while DA treatment mainly removes hemicellulose. Under the tested conditions COSLIF treatment breaks down lignocellulose structure more extensively than DA treatment, producing a more enzymatically reactive material with a higher CAC accompanied by faster hydrolysis rates and higher enzymatic digestibility.

  16. Comparative study of corn stover pretreated by dilute acid and cellulose solvent-based lignocellulose fractionation: Enzymatic hydrolysis, supramolecular structure, and substrate accessibility.

    PubMed

    Zhu, Zhiguang; Sathitsuksanoh, Noppadon; Vinzant, Todd; Schell, Daniel J; McMillan, James D; Zhang, Y-H Percival

    2009-07-01

    Liberation of fermentable sugars from recalcitrant biomass is among the most costly steps for emerging cellulosic ethanol production. Here we compared two pretreatment methods (dilute acid, DA, and cellulose solvent and organic solvent lignocellulose fractionation, COSLIF) for corn stover. At a high cellulase loading [15 filter paper units (FPUs) or 12.3 mg cellulase per gram of glucan], glucan digestibilities of the corn stover pretreated by DA and COSLIF were 84% at hour 72 and 97% at hour 24, respectively. At a low cellulase loading (5 FPUs per gram of glucan), digestibility remained as high as 93% at hour 24 for the COSLIF-pretreated corn stover but reached only approximately 60% for the DA-pretreated biomass. Quantitative determinations of total substrate accessibility to cellulase (TSAC), cellulose accessibility to cellulase (CAC), and non-cellulose accessibility to cellulase (NCAC) based on adsorption of a non-hydrolytic recombinant protein TGC were measured for the first time. The COSLIF-pretreated corn stover had a CAC of 11.57 m(2)/g, nearly twice that of the DA-pretreated biomass (5.89 m(2)/g). These results, along with scanning electron microscopy images showing dramatic structural differences between the DA- and COSLIF-pretreated samples, suggest that COSLIF treatment disrupts microfibrillar structures within biomass while DA treatment mainly removes hemicellulose. Under the tested conditions COSLIF treatment breaks down lignocellulose structure more extensively than DA treatment, producing a more enzymatically reactive material with a higher CAC accompanied by faster hydrolysis rates and higher enzymatic digestibility.

  17. Human skin cell fractions fail to self-organize within a gellan gum/hyaluronic acid matrix but positively influence early wound healing.

    PubMed

    Cerqueira, Mariana T; da Silva, Lucília P; Santos, Tírcia C; Pirraco, Rogério P; Correlo, Vitor M; Marques, Alexandra P; Reis, Rui L

    2014-05-01

    Split-thickness autografts still are the current gold standard to treat skin, upon severe injuries. Nonetheless, autografts are dependent on donor site availability and often associated to poor quality neoskin. The generation of dermal-epidermal substitutes by tissue engineering is seen as a promising strategy to overcome this problematic. However, solutions that can be safely and conveniently transplanted in one single surgical intervention are still very challenging as their production normally requires long culture time, and graft survival is many times compromised by delayed vascularization upon transplantation. This work intended to propose a strategy that circumvents the prolonged and laborious preparation period of skin substitutes and allows skin cells self-organization toward improved healing. Human dermal/epidermal cell fractions were entrapped directly from isolation within a gellan gum/hyaluronic acid (GG-HA) spongy-like hydrogel formed from an off-the-shelf dried polymeric network. Upon transplantation into full-thickness mice wounds, the proposed constructs accelerated the wound closure rate and re-epithelialization, as well as tissue neovascularization. A synergistic effect of the GG-HA matrix and the transplanted cells over those processes was demonstrated at early time points. Despite the human-derived and chimeric blood vessels found, the proposed matrix did not succeed in prolonging cells residence time and in sustaining the self-organization of transplanted human cells possibly due to primitive degradation. Despite this, the herein proposed approach open the opportunity to tackle wound healing at early stages contributing to re-epithelialization and neovascularization.

  18. Fraction Reduction through Continued Fractions

    ERIC Educational Resources Information Center

    Carley, Holly

    2011-01-01

    This article presents a method of reducing fractions without factoring. The ideas presented may be useful as a project for motivated students in an undergraduate number theory course. The discussion is related to the Euclidean Algorithm and its variations may lead to projects or early examples involving efficiency of an algorithm.

  19. Isotope fractionation

    NASA Astrophysics Data System (ADS)

    Bell, Peter M.

    A rash of new controversy has emerged around the subject of mass-independent isotope fractionation effects, particularly in the case of the oxygen isotopes. To be sure, the controversy has been around for awhile, but it has been given new impetus by the results of a recent study by Mark H. Thiemens and John E. Heidenreich III of the University of California, San Diego (Science, March 4, 1983).Gustav Arrhenius has been trying to convince the planetary science community that chemical effects in isotope fractionation processes could explain observations in meteorites that appear to be outside of the traditionally understood mass-dependent fractionations (G. Arrhenius, J . L. McCrumb, and N. F. Friedman, Astrophys. Space Sci, 65, 297, 1974). Robert Clayton had made the basic observations of oxygen in carbonaceous chondrites that the slope of the δ17 versus δ18 line was 1 instead of the slope of ½ characteristic of terrestrial rocks and lunar samples (Ann. Rev. Nucl. Part. Sci., 28, 501, 1978). The mass-independent effects were ascribed to the apparent contribution of an ancient presolar system component of O16.

  20. Activation of clones producing self-reactive antibodies by foreign antigen and antiidiotype antibody carrying the internal image of the antigen.

    PubMed Central

    Bailey, N C; Fidanza, V; Mayer, R; Mazza, G; Fougereau, M; Bona, C

    1989-01-01

    Because we found in previous work that a high fraction of antibodies exhibiting various specificities bound to glutamic acid 50-tyrosine50 homopolymer (GT) and expressed pGAT cross-reactive idiotype (IdX), we studied the activation of clones producing multireactive antibodies in 1-mo-old MRL/lpr and C3H/HeJ mice bearing VHJ haplotype. The activation of such clones was studied after mice were immunized with GT in CFA, HP20 (an anti-Id MAb carrying the internal image of GT in the D region), and a synthetic peptide corresponding to the D segment of HP20. Our results indicate that immunized mice produced both GT- and self-reactive antibodies. Study of the immunochemical properties of MAb showed that they exhibit multispecific properties and bind with similar-affinity constants to GT or self-antigens such as DNA, Smith antigen (Sm), and IgG2a. An important fraction of antibodies obtained from MRL/lpr mice immunized with HP20 expressed pGAT IdX and some of these antibodies share IdX expressed on anti-DNA, Sm, and rheumatoid factor (RFs) antibodies. The hybridomas producing multispecific autoantibodies use heavy-chain- (VH) and light-chain-variable region (VK) genes from various V gene families, suggesting that they do not derive from the pool of GAT precursors. Sequencing of VH and VK genes of two antibodies show that they can use closely related VHJ558, unmutated VK1, or different VK genes than those used by anti-GT antibodies. Our data demonstrate that clones producing antibodies binding to GT and self-antigens with similar-affinity constants can be activated by foreign or anti-Id antibodies carrying the internal image of the antigen or even by a synthetic peptide corresponding to the D segment of anti-Id antibodies. Images PMID:2760212

  1. Synthetic antigens reveal dynamics of BCR endocytosis during inhibitory signaling.

    PubMed

    Courtney, Adam H; Bennett, Nitasha R; Zwick, Daniel B; Hudon, Jonathan; Kiessling, Laura L

    2014-01-17

    B cells detect foreign antigens through their B cell antigen receptor (BCR). The BCR, when engaged by antigen, initiates a signaling cascade. Concurrent with signaling is endocytosis of the BCR complex, which acts to downregulate signaling and facilitate uptake of antigen for processing and display on the cell surface. The relationship between signaling and BCR endocytosis is poorly defined. Here, we explore the interplay between BCR endocytosis and antigens that either promote or inhibit B cell activation. Specifically, synthetic antigens were generated that engage the BCR alone or both the BCR and the inhibitory co-receptor CD22. The lectin CD22, a member of the Siglec family, binds sialic acid-containing glycoconjugates found on host tissues, inhibiting BCR signaling to prevent erroneous B cell activation. At low concentrations, antigens that can cocluster the BCR and CD22 promote rapid BCR endocytosis; whereas, slower endocytosis occurs with antigens that bind only the BCR. At higher antigen concentrations, rapid BCR endocytosis occurs upon treatment with either stimulatory or inhibitory antigens. Endocytosis of the BCR, in response to synthetic antigens, results in its entry into early endocytic compartments. Although the CD22-binding antigens fail to activate key regulators of antigen presentation (e.g., Syk), they also promote BCR endocytosis, indicating that inhibitory antigens can be internalized. Together, our observations support a functional role for BCR endocytosis in downregulating BCR signaling. The reduction of cell surface BCR levels in the absence of B cell activation should raise the threshold for BCR subsequent activation. The ability of the activating synthetic antigens to trigger both signaling and entry of the BCR into early endosomes suggests strategies for targeted antigen delivery.

  2. Comparing the heterogeneity of copper-binding characteristics for two different-sized soil humic acid fractions using fluorescence quenching combined with 2D-COS.

    PubMed

    Hur, Jin; Lee, Bo-Mi

    2011-01-01

    Heterogeneous distributions of copper-binding characteristics were compared for two ultrafiltered size fractions of a soil HA using fluorescence quenching combined with two-dimensional correlation spectroscopy (2D-COS). The apparent shapes of the original synchronous fluorescence spectra and the extent of the fluorescence quenching upon the addition of copper were similar for the two fractions. The stability constants calculated at their highest peaks were not significantly different. However, the 2D-COS results revealed that the fluorescence quenching behaviors were strongly affected by the associated wavelengths and the fraction's size. The spectral change preferentially occurred in the wavelength order of 467 nm → 451 nm → 357 nm for the 1-10 K fraction and of 376 nm → 464 nm for the >100 K fraction. The extent of the binding affinities exactly followed the sequential orders interpreted from the 2D-COS, and they exhibited the distinctive ranges of the logarithmic values from 5.86 to 4.91 and from 6.48 to 5.95 for the 1-10 K and the >100 K fractions, respectively. Our studies demonstrated that fluorescence quenching combined with 2D-COS could be successfully utilized to give insight into the chemical heterogeneity associated with metal-binding sites within the relatively homogeneous HA size fractions.

  3. Mechanism of formation of humus coatings on mineral surfaces 2. Attenuated total reflectance spectra of hydrophobic and hydrophilic fractions of organic acids from compost leachate on alumina

    USGS Publications Warehouse

    Wershaw, R. L.; Llaguno, E.C.; Leenheer, J.A.; Sperline, R.P.; Song, Y.

    1996-01-01

    Hydrophobic and hydrophilic fractions were isolated from a compost leachate. The adsorption isotherms of both fractions on alumina were measured by attenuated total reflectance infrared spectroscopy. The shapes of the adsorption isotherms of the two fractions were different. The isotherms for the hydrophilic fraction showed little change in surface excess with increasing solution concentration above 4 mg L-1. The isotherms for the hydrophobic fraction, on the other hand, displayed a marked increase in surface excess with increasing solution concentration. This increase is evidence for the formation of aggregates (admicelles or hemimicelles) on the alumina surface. Linear dichroism calculations indicated that more of the carboxylate groups in the adsorbed hydrophobic molecules than in the absorbed hydrophilic fraction were free to rotate. The hindered rotation of the carboxylate groups in the adsorbed hydrophilic-fraction molecules probably indicates that these groups are bound to surface aluminum ions by a bidentate mechanism in which the two oxygen atoms of a single carboxylate group bind to separate aluminum ions.

  4. HIV infection of monocytes inhibits the T-lymphocyte proliferative response to recall antigens, via production of eicosanoids.

    PubMed Central

    Foley, P; Kazazi, F; Biti, R; Sorrell, T C; Cunningham, A L

    1992-01-01

    Human monocytes infected in vitro with human immunodeficiency virus (HIV) soon after adherence to plastic substrate demonstrated a significantly decreased ability to restimulate autologous immune T-lymphocyte proliferation after exposure to soluble (tetanus toxoid) and particulate [herpes simplex virus (HSV)] antigen. Incubation with the cyclo-oxygenase inhibitor, indomethacin (2-5 microM), prevented inhibition of antigen-stimulated lymphocyte proliferation. The inhibitory activity was identified in ultrafiltrates containing the low molecular weight fraction (less than 3000 MW) of supernatants from HIV-infected monocyte cultures. This activity was significantly and markedly reduced in similar ultrafiltrates prepared from indomethacin-treated cultures. Increased concentrations of prostaglandin E2 (PGE2) were detected in ultrafiltrates from HIV-infected monocyte cultures compared with uninfected cultures and cultures preincubated with indomethacin. Ultrafiltrates were inhibitory when added during the presentation of antigen to T lymphocytes but not when removed from monocyte cultures prior to the addition of lymphocytes. In addition, ultrafiltrates inhibited antigen-stimulated lymphocyte proliferation and PHA-induced lymphocyte proliferation to the same extent. These data indicate that cyclo-oxygenase products of arachidonic acid, including PGE2, are produced in excess by HIV-infected monocytes and that PGE2 and perhaps other cyclo-oxygenase products are implicated in the inhibition of antigen-stimulated lymphocyte proliferation via a direct effect on T lymphocytes. PMID:1572689

  5. Molecular characterization of MT3 antigens by two-dimensional gel electrophoresis, NH2-terminal amino acid sequence analysis, and southern blot analysis.

    PubMed Central

    Sorrentino, R; Lillie, J; Strominger, J L

    1985-01-01

    The monoclonal antibody 109d6, which recognizes major histocompatibility antigen MT3-like serologic determinants, has been used to characterize the molecules bearing this determinant in HLA-DR4 and -DR7 homozygous cell lines by two-dimensional gel and sequencing analyses. By these two criteria, these molecules are identical to each other. Southern blot analysis of genomic DNA from HLA-DR1 through -DR7 homozygous cell lines with DR beta-chain gene probes reveals a striking similarity in the pattern of hybridizing fragments between DR4 and DR7 haplotypes and among DR3, DR5, and DRw6 haplotypes reminiscent of the MT3/MT2 allodeterminant distribution. The sharing of the MT2 determinant between DR3, DR5, and DRw6 haplotypes and of the MT3 determinant between DR4 and DR7 haplotypes is part of a broader "homology," which may be a consequence of more recent separation of the haplotypes sharing the MT2 determinant on the one hand and the haplotypes sharing the MT3 determinant on the other hand. Images PMID:2582424

  6. Suppression of antigen-specific antibody responses in mice exposed to perfluorooctanoic acid: Role of PPARalpha and T- and B-cell targeting

    EPA Science Inventory

    T-cell-dependent antibody responses (TDAR) are suppressed in female C57BL/6N mice exposed to ≥3.75 mg/kg of perfluorooctanoic acid (PFOA) for 15 days. To determine if suppression of humoral immunity by PFOA is peroxisome proliferator activated receptor alpha (PPARa)-dependent and...

  7. Interaction of bentonite colloids with Cs, Eu, Th and U in presence of humic acid: A flow field-flow fractionation study

    NASA Astrophysics Data System (ADS)

    Bouby, Muriel; Geckeis, Horst; Lützenkirchen, Johannes; Mihai, Silvia; Schäfer, Thorsten

    2011-07-01

    The interaction of Cs(I), Eu(III), Th(IV) and U(VI) with montmorillonite colloids was investigated in natural Grimsel Test Site groundwater over a 3 years period. The asymmetric flow field-flow fractionation combined with various detectors was applied to study size variations of colloids and to monitor colloid association of trace metals. The colloids suspended directly in the low ionic strength ( I), slightly alkaline granitic groundwater ( I = 10 -3 mol/L, pH 9.6) showed a gradual agglomeration with a size distribution shift from initially 10-200 nm to 50-400 nm within over 3 years. The Ca 2+ concentration of 2.1 × 10 -4 mol/L in the ground water is believed to be responsible for the slow agglomeration due to Ca 2+ ion exchange against Li + and Na + at the permanently charged basal clay planes. Furthermore, the Ca 2+ concentration lies close to the critical coagulation concentration (CCC) of 10 -3 mol L -1 for clay colloids. Slow destabilization may delimit clay colloid migration in this specific groundwater over long time scales. Eu(III) and Th(IV) are found predominantly bound to clay colloids, while U(VI) prevails as the UO 2(OH) 3- complex and Cs(I) remains mainly as aquo ion under our experimental conditions. Speciation calculations qualitatively represent the experimental data. A focus was set on the reversibility of metal ion-colloid binding. Addition of humic acid as a competing ligand induces rapid metal ion dissociation from clay colloids in the case of Eu(III) even after previous aging for about 3 years. Interestingly only partial dissociation occurs in the case of Th(IV). Experiments and calculations prove that the humate complexes dominate the speciation of all metal ions under given conditions. The partial irreversibility of clay bound Th(IV) is presently not understood but might play an important role for the colloid-mediated transport of polyvalent actinides over wide distances in natural groundwater.

  8. Chemical composition and acidity of size-fractionated inorganic aerosols of 2013-14 winter haze in Shanghai and associated health risk of toxic elements

    NASA Astrophysics Data System (ADS)

    Behera, Sailesh N.; Cheng, Jinping; Huang, Xian; Zhu, Qiongyu; Liu, Ping; Balasubramanian, Rajasekhar

    2015-12-01

    The severe winter haze episode that occurred in Shanghai from December 2013 to January 2014, characterized by elevated levels of particulate matter (PM), received considerable international attention because of its impacts on public health and disruption of day-to-day activities. To examine the characteristics of PM during this haze episode and to assess the chemistry behind formation of secondary inorganic aerosols (SIA) and associated health impacts due to exposure of toxic elements, we characterized eight water soluble inorganic (WSI) ions and twenty four trace elements in twelve size-fractionated PM (10 nm-9.9 μm). The average mass concentrations of coarse (1.8 μm < Dp < 9.9 μm), fine (Dp < 2.5 μm), ultrafine (0.01 μm < Dp < 0.10 μm) and nano (0.01 μm < Dp < 0.056 μm) particles during hazy days were 2.8, 5.2, 5.3 and 5.1 times higher than those during non-hazy days, respectively. The in-situ pH (pHIS), as predicted by the Aerosol Inorganic Model (AIM-IV) in all sizes of PM, was observed to be lower during hazy days (average of -0.64) than that during non-hazy days (average of -0.29); there was an increased acidity in haze aerosols. Based on the measured concentrations of particulate-bound toxic elements, health risk assessment was conducted, which revealed that the excess lifetime carcinogenic risk to individuals exposed to fine particles under haze events increased significantly (P < 0.05) to 69 ± 18 × 10-6 compared to non-hazy days (34 ± 10 × 10-6). The qualitative source attribution analysis suggested that the occurrence of haze could be due to a combination of increased emissions of PM from multiple anthropogenic sources followed by its accumulation under unfavourable meteorological conditions with lower mixing heights and less wind speeds and the formation of secondary aerosols.

  9. Transcutaneous antigen delivery system

    PubMed Central

    Lee, Mi-Young; Shin, Meong-Cheol; Yang, Victor C.

    2013-01-01

    Transcutaneous immunization refers to the topical application of antigens onto the epidermis. Transcutaneous immunization targeting the Langerhans cells of the skin has received much attention due to its safe, needle-free, and noninvasive antigen delivery. The skin has important immunological functions with unique roles for antigen-presenting cells such as epidermal Langerhans cells and dermal dendritic cells. In recent years, novel vaccine delivery strategies have continually been developed; however, transcutaneous immunization has not yet been fully exploited due to the penetration barrier represented by the stratum corneum, which inhibits the transport of antigens and adjuvants. Herein we review recent achievements in transcutaneous immunization, focusing on the various strategies for the enhancement of antigen delivery and vaccination efficacy. [BMB Reports 2013; 46(1): 17-24] PMID:23351379

  10. Tannic acid, a higher galloylated pentagalloylglucose, suppresses antigen-specific IgE production by inhibiting ɛ germline transcription induced by STAT6 activation.

    PubMed

    Kim, Yoon Hee; Yoshimoto, Miki; Nakayama, Kazuko; Tanino, Sousuke; Fujimura, Yoshinori; Yamada, Koji; Tachibana, Hirofumi

    2013-01-01

    Interleukin (IL)-4 is a critical stimulator that induces ɛ germline transcripts (ɛGT) for switch recombination to initiate immunoglobulin (Ig) E and is important in allergic disease pathogenesis. We found pentagalloylglucose (PGG) inhibited IL-4-induced ɛGT expression. PGG exerted its inhibitory function by suppressing IL-4-induced activation of IL-4Rα, JAK3 and STAT6. Furthermore, tannic acid, a higher galloylated PGG, attenuated ovalbumin-induced IgE production in vivo by inhibiting IL-4-induced ɛGT expression and the IL-4 signaling pathway. In conclusion, our results suggest that tannic acid may attenuate allergic diseases by suppressing IgE production by inhibiting IL-4-induced signaling.

  11. Characterization of the fertilization antigen 1 for the development of a contraceptive vaccine.

    PubMed Central

    Naz, R K; Phillips, T M; Rosenblum, B B

    1986-01-01

    A fertilization antigen, FA-1, was purified from either deoxycholate- or lithium diiodosalicylate-solubilized murine testes by immunoaffinity chromatography using a monoclonal antibody, MA-24, which inhibited fertilization in vitro. The FA-1 was recovered at high (11.4) or low (2.8) pH using stepwise elution procedures of the deoxycholate or lithium diiodosalicylate extracts, respectively. Both of these fractions showed a single band of 47 kDa when analyzed by NaDodSO4/PAGE and silver staining. Following removal of the detergent and extensive dialysis at pH 5.8 or treatment with 0.15 M NaCl, even in the presence of detergent, a monomer of 23 kDa was detected. Two-dimensional PAGE of FA-1 showed, four or five polypeptides in the 47-kDa or 23-kDa range. The dialyzed FA-1 contained a major 23-kDa and a minor 48-kDa band when separated on both sucrose and cesium chloride gradients. High performance size-exclusion chromatography showed a major peak at 23 kDa and a minor peak at 50 kDa. Further analysis of the 23-kDa peak by reverse-phase chromatography resolved the antigen into three peaks, which gave similar two-dimensional gel patterns as the native FA-1. Lectin affinity chromatography on a lens culinaris column demonstrated that a part of the antigen was bound to the lectin while the rest was not. The FA-1 revealed a positive reaction with periodic-Schiff reagent and contained glucose and mannose, which together constituted 18.8% of the total antigen mass. Amino acid analysis showed a high percentage of aspartic acid, glutamic acid, serine, and glycine. As a single injection of MA-24 significantly reduced fertilization rates in vivo, the purified FA-1 is an attractive candidate for the development of contraceptive vaccine. Images PMID:3461457

  12. Induction of anti-melanoma CTL response using DC transfected with mutated mRNA encoding full-length Melan-A/MART-1 antigen with an A27L amino acid substitution.

    PubMed

    Abdel-Wahab, Zeinab; Kalady, Matthew F; Emani, Sirisha; Onaitis, Mark W; Abdel-Wahab, Omar I; Cisco, Robin; Wheless, Lee; Cheng, Tsung-Yen; Tyler, Douglas S; Pruitt, Scott K

    2003-08-01

    Modification of the parental immunodominant Melan-A/MART-1 peptide (MART-1(26-35)) by replacing the alanine with leucine (A27L) enhances its immunogenicity. Because of the reported advantages of RNA over peptides in DC vaccines, we sought to mutate the MART-1 gene to encode a full-length MART-1 antigen with an A27L amino acid substitution. Human DC were transfected with A27L-mutated MART-1 RNA (A27L RNA) or native MART-1 RNA, and then used to stimulate autologous T cells from a series of 8 HLA-A2+ volunteers. After three stimulations, all CTL induced with DC/A27L RNA exhibited more tetramer+ cells, and demonstrated stronger antigen-specific IFNgamma-secreting activity compared to CTL induced with DC/native RNA. A potent MART-1-specific, and predominantly class-I-restricted lysis was detected in most CTL induced with DC/A27L RNA, while native RNA-induced CTL showed minimal and non-specific lysis. HLA-A2+ DC and MART-1 negative/A2+ melanoma cells transfected with the A27L RNA were recognized and killed by MART-1-specific CTL, suggesting that these APC efficiently processed the A27L RNA and presented correct MART-1-specific epitope(s). In summary, introducing an A27L mutation into the MART-1 full-length mRNA sequence enhanced the immunogenicity of the encoded MART-1 Ag. The ease with which such a mutation can be made in RNA presents another potential advantage of using RNA for immunotherapy. Our results support considering this strategy for enhancing the immunogenicity of DC-based RNA vaccines.

  13. Fatty acid composition in major depression: decreased omega 3 fractions in cholesteryl esters and increased C20: 4 omega 6/C20:5 omega 3 ratio in cholesteryl esters and phospholipids.

    PubMed

    Maes, M; Smith, R; Christophe, A; Cosyns, P; Desnyder, R; Meltzer, H

    1996-04-26

    Recently, there were some reports that major depression may be accompanied by alterations in serum total cholesterol, cholesterol ester and omega 3 essential fatty acid levels and by an increased C20: 4 omega 6/C20: 5 omega 3, i.e., arachidonic acid/eicosapentaenoic, ratio. The present study aimed to examine fatty acid composition of serum cholesteryl esters and phospholipids in 36 major depressed, 14 minor depressed and 24 normal subjects. Individual saturated (e.g., C14:0; C16:0, C18:0) and unsaturated (e.g., C18:1, C18:2, C20:4) fatty acids in phospholipid and cholesteryl ester fractions were assayed and the sums of the percentages of omega 6 and omega 3, saturated, branched chain and odd chain fatty acids, monoenes as well as the ratios omega 6/omega 3 and C20:4 omega 6/C20:5 omega 3 were calculated. Major depressed subjects had significantly higher C20:4 omega 6/C20:5 omega 3 ratio in both serum cholesteryl esters and phospholipids and a significantly increased omega 6/omega 3 ratio in cholesteryl ester fraction than healthy volunteers and minor depressed subjects. Major depressed subjects had significantly lower C18:3 omega 3 in cholesteryl esters than normal controls. Major depressed subjects showed significantly lower total omega 3 polyunsaturated fatty acids in cholesteryl esters and significantly lower C20:5 omega 3 in serum cholesteryl esters and phospholipids than minor depressed subjects and healthy controls. These findings suggest an abnormal intake or metabolism of essential fatty acids in conjunction with decreased formation of cholesteryl esters in major depression.

  14. Docosahexaenoic acid-induced amelioration on impairment of memory learning in amyloid beta-infused rats relates to the decreases of amyloid beta and cholesterol levels in detergent-insoluble membrane fractions.

    PubMed

    Hashimoto, Michio; Hossain, Shahdat; Agdul, Haqu; Shido, Osamu

    2005-12-30

    We investigated the effects of dietary administration of docosahexaenoic acid (DHA; C22:6n-3) on the levels of amyloid beta (A beta) peptide (1-40) and cholesterol in the nonionic detergent Triton 100 x-insoluble membrane fractions (DIFs) of the cerebral cortex and, also, on learning-related memory in an animal model of Alzheimer's disease (AD) rats infused with A beta peptide (1-40) into the cerebral ventricle. The infusion increased the levels of A beta peptide and cholesterol in the DIFs concurrently with a significant increase in reference memory errors (measured by eight-arm radial-maze tasks) compared with those of vehicle rats. Conversely, the dietary administration of DHA to AD-model rats decreased the levels of A beta peptide and cholesterol in the DIFs, with the decrease being more prominent in the DHA-administered rats. Regression analysis revealed a significant positive correlation between A beta peptide and each of cholesterol, palmitic acid and stearic acid, and between the number of reference memory errors and each of cholesterol, palmitic, stearic and oleic acid; moreover, a significant negative correlation was observed between the number of reference memory errors and the molar ratio of DHA to palmitic plus stearic acid. These results suggest that DHA-induced protection of memory deficits in AD-model rats is related to the interactions of cholesterol, palmitic acid or stearic acid with A beta peptides in DIFs where DHA ameliorates these interactions.

  15. Purification and characterization of the major antigen WI-1 from Blastomyces dermatitidis yeasts and immunological comparison with A antigen.

    PubMed Central

    Klein, B S; Jones, J M

    1994-01-01

    The lack of well-defined antigens from Blastomyces dermatitidis has hampered the ability to reliably diagnose human infection and study the immunobiology of blastomycosis. We recently discovered a novel surface protein on B. dermatitidis yeasts, designated WI-1, and demonstrated it to be a key antigenic target of humoral and cellular responses during infection. In the present article, we purified and characterized WI-1 and compared it immunologically with the only Blastomyces antigen commercially available, A antigen. WI-1 was purified by high-performance liquid chromatography over a DEAE-cellulose column. It eluted from the column at a point on the salt gradient corresponding to 460 to 490 mM NaCl, reflecting its acidic pI of approximately equal to 5.2. Purified WI-1 had a molecular mass of 120 kDa and contained a large amount of cysteine (85 residues) and aromatic amino acids but undetectable carbohydrate. In contrast, A antigen had a molecular mass of 135 kDa and contained 37% carbohydrate. Immunological comparison of the two antigens showed that, when radiolabeled, WI-1 was more reactive with anti-Blastomyces antisera than A antigen but did not cross-react with anti-Histoplasma antisera. Proteinase digestion of WI-1 eliminated its recognition by anti-WI-1 and anti-Blastomyces antisera. Proteinase treatment of A antigen had no effect on its recognition by anti-Blastomyces or anti-Histoplasma antisera, but periodate treatment abolished recognition by anti-Histoplasma antisera, indicating that the cross-reactive determinant(s) of A antigen is displayed on the accompanying carbohydrate. In further studies, anti-WI-1 antiserum reacted with A antigen and, conversely, anti-A antiserum and monoclonal antibodies (MAbs) reacted with WI-1, indicating a shared determinant on the two antigens. A recombinant 25-amino-acid repeat, recently cloned from WI-1 and found to be the major target of antibody recognition of WI-1, reacted strongly with anti-A antiserum and MAbs. In MAb

  16. The immunodominant outer membrane antigen of Actinobacillus actinomycetemcomitans is located in the serotype-specific high-molecular-mass carbohydrate moiety of lipopolysaccharide.

    PubMed Central

    Page, R C; Sims, T J; Engel, L D; Moncla, B J; Bainbridge, B; Stray, J; Darveau, R P

    1991-01-01

    Most patients with juvenile periodontitis manifest serum antibodies, sometimes at very high titers, to antigens of Actinobacillus actinomycetemcomitans, but the antigens inducing the immune response have been only partly characterized. We separated A. actinomycetemcomitans serotype b cells into protein, lipopolysaccharide (LPS), and soluble polysaccharide fractions and characterized them. Coomassie blue- and silver-stained sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels were used to detect protein and LPS components, and gas-liquid chromatography was used to determine their carbohydrate and fatty acid composition. Western blots, dot blots, and enzyme-linked immunosorbent assay inhibition with high-titer sera from juvenile periodontitis patients revealed which components were highest in antibody binding activity. These results showed that the major portion of the immunoglobulin G binding activity resides in the purified mannan-free LPS, with lesser amounts in the total protein fraction. Using Sephacryl S-300 chromatography, we separated LPS into high-molecular-mass components with high carbohydrate contents by gas-liquid chromatography and a low-molecular-mass component consisting mainly of lipid A and the inner core sugar heptulose. The results of quantitative dot blot assays and enzyme-linked immunosorbent assay inhibition show that the serotype-specific antibody binding activity is highly concentrated in the high-molecular-mass carbohydrate-rich LPS fraction and is almost completely absent in the low-molecular-weight lipid-rich fraction. Our observations contrast with previous reports that the predominant serotype antigen of A. actinomycetemcomitans resides in a mannan-rich polysaccharide isolated from spent culture medium. These observations support the conclusion that the immunodominant antigen of the outer membrane is the O antigen of the LPS. Images PMID:1716610

  17. The immunologically reactive O-linked polysaccharide chains derived from circulating cathodic antigen isolated from the human blood fluke Schistosoma mansoni have Lewis x as repeating unit.

    PubMed

    Van Dam, G J; Bergwerff, A A; Thomas-Oates, J E; Rotmans, J P; Kamerling, J P; Vliegenthart, J F; Deelder, A M

    1994-10-01

    The gut-associated excretory antigen circulating cathodic antigen (CCA) was isolated by immunoaffinity chromatography from adult Schistosoma mansoni worms, which were collected from infected golden hamsters. This antigen is probably involved in protection of the schistosome gut and is increasingly used in highly sensitive and specific immunodiagnostic assays. Amino acid analysis before and after alkaline borohydride treatment of CCA and monosaccharide analysis indicated that CCA is O-glycosylated mostly via GalNAc-Thr. After reductive alkaline treatment, the O-linked carbohydrate chains were fractionated by gel-permeation chromatography, followed by normal-phase HPLC on LiChrosorb-NH2. Carbohydrate-positive fractions were investigated by one-dimensional and two-dimensional 1H-NMR spectroscopy, fast atom bombardment mass spectrometry and collision-induced-dissociation tandem mass spectrometry. The analyses showed that the low-molecular-mass O-linked oligosaccharide alditols (the minor fraction) consist of disaccharides to hexasaccharides having the Gal beta (1-3)GalNAc-OL core in common. The major carbohydrate fraction comprises a population of polysaccharides, containing Lewis x repeating units (-3)Gal beta (1-4)[Fuc alpha (1-3)]GlcNAc beta (1-). CCA-specific monoclonal antibodies and IgM antibodies in patient sera recognized the fucosylated O-linked carbohydrate antigenic structures. Since CCA evokes a strong IgM antibody response and carbohydrate structures containing repeating Lewis x units are found on circulating neutrophils, it is proposed that the antigenic poly-Lewis x polysaccharide of CCA is involved in the induction of auto-immunity against granulocytes, resulting in the mild to moderate neutropenia observed during schistosome infection.

  18. New strategy for the determination of gliadins in maize- or rice-based foods matrix-assisted laser desorption/ionization time-of-flight mass spectrometry: fractionation of gliadins from maize or rice prolamins by acidic treatment.

    PubMed

    Hernando, Alberto; Valdes, Israel; Méndez, Enrique

    2003-08-01

    A procedure for determining small quantities of gliadins by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) in gluten-free foods containing relatively large amounts of prolamin proteins from maize or rice is described. We report for the first time that gliadins, the ethanol-soluble wheat prolamin fraction, can be quantitatively solubilized in 1.0 M acetic acid, while the corresponding ethanol-soluble maize or rice prolamin fraction remains insoluble in acetic acid. We describe a methodology for the detection of gliadins in maize and rice foods based on a two-step procedure of extraction (60% aqueous ethanol followed by 1 M acetic acid). Subsequent MALDI-TOFMS analysis of the resulting acidic extract from these gluten-free foods clearly confirms the presence of a typical mass pattern corresponding to gliadin components, ranging from 30 to 45 kDa. Depending on the percentages of maize or rice flours employed in the elaboration of these foods, the combined procedure enables levels of gliadins from 100 to 400 ppm to be detected. The efficiency of this combined procedure corroborates enzyme-linked immunosorbent assay data for a large number of maize/rice gluten-free foods by means of direct visualization of the characteristic gliadin mass pattern in maize or rice foods.

  19. [Enterobacterial antigen in human peripheral blood lymphocytes].

    PubMed

    Faure-Fontenla, M A; García-Tamayo, F

    1989-11-01

    The following study has as prior history the research reports which have shown the existence of an antigenic tissue deposit in gram-negative enterobacteria. The antigens of the enterobacteria have also been found in the lymphocytic membranes and cytoplasm. Since intestinal lymphoid tissue cells can recirculate by means of the thoracic duct to the peripheral venous system, it was proposed that the circulating lymphocytes in healthy people could also contain small amounts of a common enterobacterial antigen. The study was carried out in 15 human venous blood samples, of which the lymphocytic population was separated to later be used in the preparation of 15 alcohol soluble extracts. This material was used for inhibiting the immuno-hemolysis assay in three occasions in order to show the presence of antigens shared by different enterobacterias, using as reference a fraction separated from the LPS of Escherichia coli 08. The results showed that the human lymphocytes also had antigenic determinants common to gram-negative bacteria.

  20. Composition of the amino acid and amino sugar for molecular weight fractions of hot-water extractable soil organic matters from soils with plant residue compost or mineral fertilization

    NASA Astrophysics Data System (ADS)

    Moriizumi, M.; Matsunaga, T.; Uezono, I.; Kato, N.

    2009-04-01

    The hot-water extractable organic nitrogen is well known as a laboratory index of mineralizable nitrogen. This available nitrogen is indispensable for growth of plants because of being absorbed in crops. We measured the composition of the amino acid and amino sugar for molecular weight fractions in hot-water extractable organic matters to understand the source of the available nitrogen in soils inserted a compost. Two soil samples were collected from fields (Soil Type; Andosol) in National Agricultural Research Center in Tsukuba, Japan. A plant residue compost of 2 kga-1y-1 during 25 year has been applied to a soil and another soil was under the mineral fertilization. Organic matters were extracted from the soils of 3 g in the water of 50 ml at 80 degree centigrade for16 hours. The molecular size distribution of the hot-water extractable organic matters was analyzed by HPSCE (column YMC Diol-120, elution; 50mM phosphate buffer under pH=7.0, flow rate 1 mlmin-1), and 20 fractions were collected at regular intervals in the retention time. The chromatograms were monitored under the absorbance at 280 nm and fluorescence intensity at Ex.280 nm: Em.330nm. The concentrations of the 15 amino acids and three amino sugars (muramic acid, glucosamine, and galactosamine) for the molecule weight fractions were measured by HPLC as o-phthaldialdehyde (OPA) derivatives after the vapor HCl hydrolysis. Organic nitrogen concentrations of the hot-water extractable organic matters in the soil inserted the compost (C-soil) and the mineral fertilization soil (M-soil) were 133 and 35 mgkg-1, respectively. The extracted organic matters had the variable molecule weight (103- 104 Da). The concentrations of the amino acid and amino sugar of organic nitrogen in the C-soil were higher than those in the M-soil in all fractions. The fractions were classified into 3 groups (LW, MW, and SW) based on the molecule weight and spectroscopic characteristics. Each group had unique composition of the amino

  1. Fractionation and utilization of gossypol resin

    SciTech Connect

    Tursunov, A.K.; Dzhailov, A.T.; Fatkhullaev, E.; Sadykov, A.A.

    1985-10-01

    Gossypol resin is formed as a secondary waste product during distillation of fatty acides isolated from cottonseed oil soap stocks; it is insoluble in water but soluble in products of petroleum distillation. For fractionation, gossypol resin was saponified with caustic soda or caustic potash. Using this method, the resin was separated into unsaponifiable (21-24%) and saponifiable (76-79%) parts. Details of the individual fractions of gossypol resin are presented. The unsaponifiable fraction contains hydrocarbons, alcohols, beta-sito-sterol, beta-amyrin, and vitamin E. The fatty acid fraction of the resin is a mixture of fatty acids and lactones.

  2. Ganglioside contained in the neuronal tissue-enriched acidic protein of 22 kDa (NAP-22) fraction prepared from the detergent-resistant membrane microdomain of rat brain inhibits the phosphatase activity of calcineurin.

    PubMed

    Kobayashi, Yuumi; da Silva, Ronan; Kumanogoh, Haruko; Miyata, Shinji; Sato, Chihiro; Kitajima, Ken; Nakamura, Shun; Morita, Mistuhiro; Hayashi, Fumio; Maekawa, Shohei

    2015-09-01

    Neurons have well-developed membrane microdomains called "rafts" that are recovered as a detergent-resistant membrane microdomain fraction (DRM). Neuronal tissue-enriched acidic protein of 22 kDa (NAP-22) is one of the major protein components of neuronal DRM. To determine the cellular function of NAP-22, interacting proteins were screened with an immunoprecipitation assay, and calcineurin (CaN) was detected. Further studies with NAP-22 prepared from DRM and CaN expressed in bacteria showed the binding of these proteins and a dose-dependent inhibitory effect of the NAP-22 fraction on the phosphatase activity of CaN. On the other hand, NAP-22 expressed in bacteria showed low binding to CaN and a weak inhibitory effect on phosphatase activity. To solve this discrepancy, identification of a nonprotein component that modulates CaN activity in the DRM-derived NAP-22 fraction was attempted. After lyophilization, a lipid fraction was extracted with chloroform/methanol. The lipid fraction showed an inhibitory effect on CaN without NAP-22, and further fractionation of the extract with thin-layer chromatography showed the presence of several lipid bands having an inhibitory effect on CaN. The mobility of these bands coincided with that of authentic ganglioside (GM1a, GD1a, GD1b, and GT1b), and authentic ganglioside showed an inhibitory effect on CaN. Treatment of lipid with endoglycoceramidase, which degrades ganglioside to glycochain and ceramide, caused a diminution of the inhibitory effect. These results show that DRM-derived NAP-22 binds several lipids, including ganglioside, and that ganglioside inhibits the phosphatase activity of CaN.

  3. A randomized blinded clinical trial of two antivenoms, prepared by caprylic acid or ammonium sulphate fractionation of IgG, in Bothrops and Porthidium snake bites in Colombia: correlation between safety and biochemical characteristics of antivenoms.

    PubMed

    Otero, R; Gutiérrez, J M; Rojas, G; Núñez, V; Díaz, A; Miranda, E; Uribe, A F; Silva, J F; Ospina, J G; Medina, Y; Toro, M F; García, M E; León, G; García, M; Lizano, S; De La Torre, J; Márquez, J; Mena, Y; González, N; Arenas, L C; Puzón, A; Blanco, N; Sierra, A; Espinal, M E; Lozano, R

    1999-06-01

    A randomized blinded clinical trial was performed in 53 patients bitten by Bothrops sp. and Porthidium sp. in Antioquia and Chocó, Colombia, in order to compare the efficacy and safety of two antivenoms made of whole IgG obtained by either ammonium sulphate (monovalent anti-B. atrox) or caprylic acid (polyvalent) fractionation. Additionally, antivenoms were compared by electrophoretic and chromatographic analyses and anticomplementary activity in vitro. With a protocol of 2, 4 and 6 antivenom vials for the treatment of mild, moderate and severe envenomings, respectively, both antivenoms were equally efficient to neutralize the most relevant signs of envenoming and to clear serum venom levels in patients from the first hour and later on. Three patients with severe envenoming and initially treated with less than six vials on admission had persistent or recurrent venom antigenemia within 12-48 h. Monovalent antivenom fractionated by ammonium sulphate precipitation had higher amounts of protein aggregates and nonimmunoglobulin proteins than polyvalent antivenom fractionated by caprylic acid precipitation. Both antivenoms presented anticomplementary activity in vitro, being higher in the monovalent product. In agreement, monovalent antivenom induced a significantly higher incidence of early antivenom reactions (52%) than polyvalent antivenom (25%).

  4. Antigen injection (image)

    MedlinePlus

    Leprosy is caused by the organism Mycobacterium leprae . The leprosy test involves injection of an antigen just under ... if your body has a current or recent leprosy infection. The injection site is labeled and examined ...

  5. Effect of calcium on fatty acid bioaccessibility during in vitro digestion of Cheddar-type cheeses prepared with different milk fat fractions.

    PubMed

    Ayala-Bribiesca, Erik; Turgeon, Sylvie L; Britten, Michel

    2017-04-01

    Calcium plays an important role in intestinal lipid digestion by increasing the lipolysis rate, but also limits fatty acid bioaccessibility by producing insoluble Ca soaps with long-chain fatty acids at intestinal pH conditions. The aim of this study was to better understand the effect of Ca on the bioaccessibility of milk fat from Cheddar-type cheeses. Three anhydrous milk fats (AMF) with different fatty acid profiles (olein, stearin, or control AMF) were used to prepare Cheddar-type cheeses, which were then enriched or not with Ca using CaCl2 during the salting step. The cheeses were digested in vitro, and their disintegration and lipolysis rates were monitored during the process. At the end of digestion, lipids were extracted under neutral and acidic pH conditions to compare free fatty acids under intestinal conditions in relation to total fatty acids released during the digestion process. The cheeses prepared with the stearin (the AMF with the highest ratio of long-chain fatty acids) were more resistant to disintegration than the other cheeses, owing to the high melting temperature of that AMF. The Ca-enriched cheeses had faster lipolysis rates than the regular Ca cheeses. Chromatographic analysis of the digestion products showed that Ca interacted with long-chain fatty acids, producing Ca soaps, whereas no interaction with shorter fatty acids was detected. Although higher Ca levels resulted in faster lipolysis rates, driven by the depletion of reaction products as Ca soaps, such insoluble compounds are expected to reduce the bioavailability of fatty acids by hindering their absorption. These effects on lipid digestion and absorption are of interest for the design of food matrices for the controlled release of fat-soluble nutrients or bioactive molecules.

  6. A universal computational model for predicting antigenic variants of influenza A virus based on conserved antigenic structures

    PubMed Central

    Peng, Yousong; Wang, Dayan; Wang, Jianhong; Li, Kenli; Tan, Zhongyang; Shu, Yuelong; Jiang, Taijiao

    2017-01-01

    Rapid determination of the antigenicity of influenza A virus could help identify the antigenic variants in time. Currently, there is a lack of computational models for predicting antigenic variants of some common hemagglutinin (HA) subtypes of influenza A viruses. By means of sequence analysis, we demonstrate here that multiple HA subtypes of influenza A virus undergo similar mutation patterns of HA1 protein (the immunogenic part of HA). Further analysis on the antigenic variation of influenza A virus H1N1, H3N2 and H5N1 showed that the amino acid residues’ contribution to antigenic variation highly differed in these subtypes, while the regional bands, defined based on their distance to the top of HA1, played conserved roles in antigenic variation of these subtypes. Moreover, the computational models for predicting antigenic variants based on regional bands performed much better in the testing HA subtype than those did based on amino acid residues. Therefore, a universal computational model, named PREDAV-FluA, was built based on the regional bands to predict the antigenic variants for all HA subtypes of influenza A viruses. The model achieved an accuracy of 0.77 when tested with avian influenza H9N2 viruses. It may help for rapid identification of antigenic variants in influenza surveillance. PMID:28165025

  7. Isolation and purification of antigenic components of Cryptococcus.

    PubMed

    Wozniak, Karen L; Levitz, Stuart M

    2009-01-01

    The encapsulated fungal pathogens Cryptococcus neoformans and Cryptococcus gattii are significant agents of life-threatening infections, particularly in persons with suppressed cell-mediated immunity. This chapter provides detailed methodology for the purification of two of the major antigen fractions of C. neoformans: glucuronoxylomannan (GXM) and mannoprotein (MP). GXM is the primary component of the polysaccharide capsule, which is the major cryptococcal virulence factor. In contrast, MPs have been identified as key antigens that stimulate T-cell responses. Purification of GXM and MP should assist investigators studying the antigenic, biochemical, and virulence properties of Cryptococcus species.

  8. Human immunodeficiency virus contains an epitope immunoreactive with thymosin. cap alpha. /sub 1/ and the 30-amino acid synthetic p17 group-specific antigen peptide HGP-30

    SciTech Connect

    Naylor, P.H.; Naylor, C.W.; Badamchian, M.; Wada, S.; Goldstein, A.L.; Wang, S.S.; Sun, D.K.; Thornton, A.H.; Sarin, P.S.

    1987-05-01

    The authors have reported that an antiserum prepared against thymosin ..cap alpha../sub 1/ (which shares a region of homology with the p17 protein of the acquired immunodeficiency syndrome (AIDS)-associated human immunodeficiency virus) effectively neutralized the AIDs virus and prevented its replication in H9 cells. Using HPLC and immunoblot analysis, they have identified from a clone B, type III human T-lymphotropic virus (HTLV-IIIB) extracts a protein with a molecular weight of 17,000 that is immunoreactive with thymosin ..cap alpha../sub 1/. In contrast, no immunoreactivity was found in retroviral extracts from a number of nonhuman species including feline, bovine, simian, gibbon, and murine retroviruses. Heterologous antiserum prepared against a 30-amino acid synthetic peptide analogue (HGP-30) does not cross-react with thymosin ..cap alpha../sub 1/ but does react specifically with the p17 protein of the AIDS virus in a manner identical to that seen with an HTLV-IIIB p17-specific monoclonal antibody. The demonstration that this synthetic analogue is immunogenic and that antibodies to HGP-30 cross-react not only with synthetic peptide but also with the HTLV-IIIB p17 viral protein provides an additional, and potentially more specific, candidate for development of a synthetic peptide vaccine for AIDS. In addition, the p17 synthetic peptide (HGP-3) may prove to be useful in a diagnostic assay for the detection of AIDS virus infection in seronegative individuals.

  9. Distribution and linkage of domoic acid (amnesic shellfish poisoning toxins) in subcellular fractions of the digestive gland of the scallop Pecten maximus.

    PubMed

    Mauriz, Aida; Blanco, Juan

    2010-01-01

    The king scallop Pecten maximus accumulates domoic acid, the main amnesic shellfish poisoning toxin, in the digestive gland for a long time. To try to find if the cause of this characteristic is the binding of the toxin to some cellular component, the subcellular distribution of domoic acid in the cells of the digestive gland was studied, by means of serial centrifugation, ultrafiltration and size exclusion chromatography (SEC). Domoic acid was found to be present mostly in soluble form in the cytosol, as more than 90% was found in the supernatant after a centrifugation of 1h at 45,000 x g, and passed a 10 kDa ultrafilter. The retention time of the peak with an absorption maximum of 242 nm--the one characteristic of domoic acid--observed in the SEC chromatograms of the scallop samples was found identical to be one of a reference solution of the toxin, indicating therefore, that domoic acid is free in the cytosol of the digestive gland of Pecten maximus. This finding turns the focus from binding to the lack of membrane transporters in this species of the scallop as the cause of the long retention time of domoic acid in this species.

  10. The Pattern of Fatty Acids Displaced by EPA and DHA Following 12 Months Supplementation Varies between Blood Cell and Plasma Fractions.

    PubMed

    Walker, Celia G; West, Annette L; Browning, Lucy M; Madden, Jackie; Gambell, Joanna M; Jebb, Susan A; Calder, Philip C

    2015-08-03

    Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are increased in plasma lipids and blood cell membranes in response to supplementation. Whilst arachidonic acid (AA) is correspondingly decreased, the effect on other fatty acids (FA) is less well described and there may be site-specific differences. In response to 12 months EPA + DHA supplementation in doses equivalent to 0-4 portions of oily fish/week (1 portion: 3.27 g EPA+DHA) multinomial regression analysis was used to identify important FA changes for plasma phosphatidylcholine (PC), cholesteryl ester (CE) and triglyceride (TAG) and for blood mononuclear cells (MNC), red blood cells (RBC) and platelets (PLAT). Dose-dependent increases in EPA + DHA were matched by decreases in several n-6 polyunsaturated fatty acids (PUFA) in PC, CE, RBC and PLAT, but were predominantly compensated for by oleic acid in TAG. Changes were observed for all FA classes in MNC. Consequently the n-6:n-3 PUFA ratio was reduced in a dose-dependent manner in all pools after 12 months (37%-64% of placebo in the four portions group). We conclude that the profile of the FA decreased in exchange for the increase in EPA + DHA following supplementation differs by FA pool with implications for understanding the impact of n-3 PUFA on blood lipid and blood cell biology.

  11. Polyomavirus T Antigens Activate an Antiviral State

    PubMed Central

    Giacobbi, Nicholas S.; Gupta, Tushar; Coxon, Andrew; Pipas, James M.

    2014-01-01

    Ectopic expression of Simian Virus 40 (SV40) large T antigen (LT) in mouse embryonic fibroblasts (MEFs) increased levels of mRNAs encoding interferon stimulated genes (ISGs). The mechanism by which T antigen increases levels of ISGs in MEFs remains unclear. We present evidence that expression of T antigen from SV40, Human Polyomaviruses BK (BKV) or JC (JCV) upregulate production of ISGs in MEFs, and subsequently result in an antiviral state, as determined by inhibition of VSV or EMCV growth. The first 136 amino acids of LT are sufficient for these activities. Furthermore, increased ISG expression and induction of the antiviral state requires STAT1. Finally, the RB binding motif of LT is necessary for activation of STAT1. We conclude that the induction of the STAT1 mediated innate immune response in MEFs is a common feature shared by SV40, BKV and JCV. PMID:25589241

  12. IMMUNOLOGICAL ACTIVITY OF STAPHYLOCOCCUS FOR ANTI-ANTHRAX PRECIPITIN SERUM. II. ON THE COMMON ANTIGEN BETWEEN B. ANTHRACIS AND STAPHYLOCOCCUS

    DTIC Science & Technology

    and CCL3COOH precipitated fraction by absorption test and agar dispersion method. The common antigenic substance was thought to be a protein...substance having Molisch negative, Biuret positive and Ninhydrin positive reactions. No common antigenic substance was observed in the fractions A-4, and S-4

  13. [Surface-active agents from the group of polyoxyethylated glycerol esters of fatty acids. Part II. Chromatographic analysis and basic viscosity parameters as a estimate criterion of efficiency of catalytic oxyethylation of Lard's fractions (Adeps suillus FP VII)].

    PubMed

    Piotrowska, Jowita Barbara; Nachajski, Michał Jakub; Lukosek, Marek; Kosno, Jacek; Zgoda, Marian Mikołaj

    2011-01-01

    The catalytic oxyethylation products of Lard's fractions and Tweens--as a reference products, were analised by chromatographic analysis HPLC and GPC. The above part was determination average molecular weights dispersion (Mw I Mn) and the content of polyethylene glycols (PEG), which are obtained during catalytic oxyethylation, and determination iodine value of the product (L(J2)). Viscosity measurements were carried out by Ubelohde method and enabled determination of basic viscosity and hydrodynamic parameters. The obtained results indicate that, comparing reference products--polysorbates, Tweens, products of triglycerides oxyethylation contain significantly less, in some cases very small amount of polyethylene glycols (PEG), and also maintained a high amount of unsaturated fatty acids (oleic acid), which is proven by the iodine value. Numerical value n(s) /H2O/ confirms important disparity of micells palisad structure, which are created based on oxyethylated triglycerides nTE = 40. That indicates significant solubilization possibilities of their aqueous solutions Cexp < or = Cmc.

  14. Genetic and antigenic changes in porcine rubulavirus

    PubMed Central

    Sánchez-Betancourt, José I.; Trujillo, María E.; Mendoza, Susana E.; Reyes-Leyva, Julio; Alonso, Rogelio A.

    2012-01-01

    Blue eye disease, caused by a porcine rubulavirus (PoRV), is an emergent viral swine disease that has been endemic in Mexico since 1980. Atypical outbreaks were detected in 1990 and 2003. Growing and adult pigs presented neurological signs, mild neurological signs were observed in piglets, and severe reproductive problems were observed in adults. Amino acid sequence comparisons and phylogenetic analysis of the hemagglutinin-neuraminidase (HN) protein revealed genetically different lineages. We used cross-neutralization assays, with homologous and heterologous antisera, to determine the antigenic relatedness values for the PoRV isolates. We found antigenic changes among several strains and identified a highly divergent one, making up a new serogroup. It seems that genetically and antigenically different PoRV strains are circulating simultaneously in the swine population in the geographical region studied. The cross neutralization studies suggest that the HN is not the only antigenic determinant participating in the antigenic changes among the different PoRV strains. PMID:22754092

  15. Complex of simian virus 40 large tumor antigen and 48,000-dalton host tumor antigen.

    PubMed Central

    Greenspan, D S; Carroll, R B

    1981-01-01

    Simian virus 40 large tumor antigen (T Ag) can be separated by sucrose gradient sedimentation into a rapidly sedimenting, maximally phosphorylated fraction and a slowly sedimenting, less phosphorylated fraction. The Mr 48,000 host tumor antigen (48,000 HTA, also called nonviral T Ag) is preferentially complexed with the maximally phosphorylated T Ag. Pulse-labeled T Ag sediments as a 5-6S monomer, whereas T Ag radiolabeled for progressively longer periods slowly increases in sedimentation coefficient to give a broad distribution between 5 S and greater than 28 S. Mutation in the viral A locus causes a decrease in T Ag phosphorylation and a marked decrease in 48,000 HTA binding, shifting the sedimentation coefficient of T Ag to the monomer value. The more highly phosphorylated T Ag also has the highest affinity for chromatin. Images PMID:6941238

  16. Mapping of phosphorylation sites in polyomavirus large T antigen

    SciTech Connect

    Hassauer, M.; Scheidtmann, K.H.; Walter, G.

    1986-06-01

    The phosphorylation sites of polyomavirus large T antigen from infected or transformed cells were investigated. Tryptic digestion of large T antigen from infected, /sup 32/P/sub i/-labeled cells revealed seven major phosphopeptides. Five of these were phosphorylated only at serine residues, and two were phosphorylated at serine and threonine residues. The overall ratio of phosphoserine to phosphothreonine was 6:1. The transformed cell line B4 expressed two polyomavirus-specific phosphoproteins: large T antigen, which was only weakly phosphorylated, and a truncated form of large T antigen of 34,000 molecular weight which was heavily phosphorylated. Both showed phosphorylation patterns similar to that of large T antigen from infected cells. Peptide analyses of large T antigens encoded by the deletion mutants dl8 and dl23 or of specific fragments of wild-type large T antigen indicated that the phosphorylation sites are located in an amino-terminal region upstream of residue 194. The amino acid composition of the phosphopeptides as revealed by differential labeling with various amino acids indicated that several phosphopeptides contain overlapping sequences and that all phosphorylation sites are located in four tryptic peptides derived from a region between Met71 and Arg191. Two of the potential phosphorylation sites were identified as Ser81 and Thr187. The possible role of this modification of large T antigen is discussed.

  17. Kinetic and Modeling Investigation to Provide Design Guidelines for the NREL Dilute-Acid Process Aimed at Total Hydrolysis/Fractionation of Lignocellulosic Biomass: July 1998

    SciTech Connect

    Lee, Y. Y.; Iyer, P.; Xiang, Q.; Hayes, J.

    2004-08-01

    Following up on previous work, subcontractor investigated three aspects of using NREL ''pretreatment'' technology for total hydrolysis (cellulose as well as hemicellulose) of biomass. Whereas historic hydrolysis of biomass used either dilute acid or concentrated acid technology for hydrolysis of both hemicellulose and cellulose, NREL has been pursuing very dilute acid hydrolysis of hemicellulose followed by enzymatic hydrolysis of cellulose. NREL's countercurrent shrinking-bed reactor design for hemicellulose hydrolysis (pretreatment) has, however, shown promise for total hydrolysis. For the first task, subcontractor developed a mathematical model of the countercurrent shrinking bed reactor operation and, using yellow poplar sawdust as a feedstock, analyzed the effect of: initial solid feeding rate, temperature, acid concentration, acid flow rate, Peclet number (a measure of backmixing in liquid flow), and bed shrinking. For the second task, subcontractor used laboratory trials, with yellow poplar sawdust and 0.07 wt% sulfuric acid at various temperatures, to verify the hydrolysis of cellulose to glucose (desired) and decomposition of glucose (undesired) and determine appropriate parameters for use in kinetic models. Unlike cellulose and hemicellulose, lignins, the third major component of biomass, are not carbohydrates that can be broken down into component sugars. They are, however, aromatic complex amorphous phenolic polymers that can likely be converted into low-molecular weight compounds suitable for production of fuels and chemicals. Oxidative degradation is one pathway for such conversion and hydrogen peroxide would be an attractive reagent for this, as it would leave no residuals. For the third task, subcontractor reacted lignin with hydrogen peroxide under various conditions and analyzed the resulting product mix.

  18. Effects of sulphur, nitrogen, phosphorus, potassium, and water stress on dietary fibre fractions, starch, amino acids and on the biological value of potato protein.

    PubMed

    Eppendorfer, W H; Eggum, B O

    1994-06-01

    In pot experiments with greatly differing rates of N, P, K, and S, and 3 levels of water, dry matter (DM) yields of tubers varied from 28 to 454 g/pot. Especially P-, K- and S-deficiency reduced the starch content of boiled potatoes, from P from 74 to 59% in DM. S-deficiency increased soluble, insoluble and total digestible fibre (TDF) from about 9 to 12.4% TDF in DM of boiled potatoes. Lignin content of fresh potato DM was increased from 0.7 to 2.0 and from 0.8 to 3.7% by P- and K-deficiency. P-deficiency considerably increased arabinose, galactose, and uronic acid, and decreased glucose content. N-application and P-, K- and S-deficiency increased total- and NO3-N concentrations which varied from 1.32 to 3.67% and from 17 to 400 ppm in DM. Water stress slightly decreased total-N content. Increasing N in DM, due to high N-rates or P- or K-deficiency, decreased concentrations in crude protein (CP) of all essential amino acids, whereas aspartic acid (asparagine) increased. S-deficiency caused particularly strong decreases in concentrations of essential amino acids from 1.28 to 0.49, 1.62 to 1.10, 5.24 to 3.68, and 5.59 to 2.57 g/16 g N of cystine, methionine, lysine and leucine, respectively. Glutamic acid (glutamine) content was increased from 15.7 to 27.6 g/16 g N by S-deficiency. Expressed as g amino acid/kg DM, all amino acid concentrations increased with increasing % N in DM. In N-balance trials with rats, increasing crude protein concentrations in DM of boiled potatoes increased the true digestibility (TD) of the protein from 72 to 90 but decreased the biological value (BV) from 89 to 65. S-deficiency caused a further reduction of the BV to 45. Excluding S-deficiency treatments, linear regression equations between CP concentrations and BV and TD gave correlation coefficients r of -0.94*** and 0.82***, respectively. There was close agreement between changes of BV and concentrations of first limiting amino acids (chemical score), with r = 0.96***.

  19. The cell proliferation-associated antigen of antibody Ki-67: a very large, ubiquitous nuclear protein with numerous repeated elements, representing a new kind of cell cycle-maintaining proteins

    PubMed Central

    1993-01-01

    The antigen defined by mAb Ki-67 is a human nuclear protein the expression of which is strictly associated with cell proliferation and which is widely used in routine pathology as a "proliferation marker" to measure the growth fraction of cells in human tumors. Ki-67 detects a double band with apparent molecular weights of 395 and 345 kD in immunoblots of proteins from proliferating cells. We cloned and sequenced the full length cDNA, identified two differentially spliced isoforms of mRNA with open reading frames of 9,768 and 8,688 bp encoding for this cell proliferation-associated protein with calculated molecular weights of 358,761 D and 319,508 D, respectively. New mAbs against a bacterially expressed part and a synthetic polypeptide deduced from the isolated cDNA react with the native Ki-67 antigen, thus providing a circle of evidence that we have cloned the authentic Ki-67 antigen cDNA. The central part of the Ki-67 antigen cDNA contains a large 6,845-bp exon with 16 tandemly repeated 366-bp elements, the "Ki-67 repeats", each including a highly conserved new motif of 66 bp, the "Ki-67 motif", which encodes for the epitope detected by Ki-67. Computer analysis of the nucleic acid and the deduced amino acid sequence of the Ki-67 antigen confirmed that the cDNA encodes for a nuclear and short-lived protein without any significant homology to known sequences. Ki-67 antigen-specific antisense oligonucleotides inhibit the proliferation of IM-9 cell line cells, indicating that the Ki-67 antigen may be an absolute requirement for maintaining cell proliferation. We conclude that the Ki-67 antigen defines a new category of cell cycle-associated nuclear nonhistone proteins. PMID:8227122

  20. Diagnostic Antigens of Leishmania.

    DTIC Science & Technology

    1994-01-31

    braziliensis (MHOM/BR/75/M2903), L. chagasi (MJOM/BR/82/BA-2,C 1), L. donovani (MHOMiEt/67iHU3), Leishmania guyanensis (MIHOMJBR/75/M4147), L. infantum (IPT-1...comparative test to a variety of other recombinant Leishmania antigens including L. chagasi hsp70, L. braziliensis hsp83/90, L. braziliensis eIF4A, L...34 4. AD CONTRACT NO: DAMD17-92-C-2082 EC•£ 2 j 994 ’i, L TITLE: DIAGNOSTIC ANTIGENS OF LEISHMANIA L PRINCIPAL INVESTIGATOR: Steven G. Reed, Ph.D

  1. Heterogeneous uptake of gaseous nitric acid on dolomite (CaMg(CO3)2) and calcite (CaCO3) particles: a Knudsen cell study using multiple, single, and fractional particle layers.

    PubMed

    Johnson, Elizabeth R; Sciegienka, Joanna; Carlos-Cuellar, Sofia; Grassian, Vicki H

    2005-08-11

    In this study, the heterogeneous uptake of gaseous nitric acid on dolomite, CaMg(CO3)2, and calcite, CaCO3, particles under dry conditions at 296 K was investigated. A Knudsen cell reactor was used to measure heterogeneous uptake coefficients for these reactions. Several different experiments were performed including those on many, single, and fractional layers of particles. For experiments using multiple particle layers, the Knudsen cell data were modeled to take into account gas diffusion into the underlying layers of the sample. From this analysis, initial heterogeneous uptake coefficients, gamma(o,t), were determined to be (5 +/- 2) x 10(-4) and (2 +/- 1) x 10(-3), for dolomite and calcite, respectively, at a nitric acid concentration of 6.5 x 10(10) molecules cm(-3). For experiments that employed single or fractional particle layers, the initial heterogeneous uptake coefficient was analyzed using a recent method described in the literature. Values of the initial heterogeneous uptake coefficient using this analysis were in agreement with the above analysis and determined to be (7 +/- 4) x 10(-4) and (2 +/- 0.4) x 10(-3) for CaMg(CO3)2 and CaCO3, respectively. In addition, these results are compared to previous literature values.

  2. A substantial fraction of barley (Hordeum vulgare L.) low phytic acid mutations have little or no effect on yield across diverse production environments

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The potential benefits of the low phytic acid (lpa) seed trait for human and animal nutrition, and for phosphorus management in non-ruminant animal production, are well documented. However, in many cases the lpa trait is associated with impaired seed or plant performance, resulting in reduced yield....

  3. Carbon-Isotope Composition of Biochemical Fractions and the Regulation of Carbon Balance in Leaves of the C3-Crassulacean Acid Metabolism Intermediate Clusia minor L. Growing in Trinidad.

    PubMed

    Borland, A. M.; Griffiths, H.; Broadmeadow, MSJ.; Fordham, M. C.; Maxwell, C.

    1994-10-01

    Carbon-isotope ratios ([delta]13Cs) were measured for various bio-chemical fractions quantitatively extracted from naturally exposed and shaded leaves of the C3-Crassulacean acid metabolism (CAM) intermediate Clusia minor, sampled at dawn and dusk on days during the wet and dry seasons in Trinidad. As the activity of CAM increased in response to decreased availability of water and higher photon flux density, organic acids and soluble sugars were enriched in 13C by approximately 3.5 to 4%[per mille (thousand) sign] compared to plants sampled during the wet season. The induction of CAM was accompanied by a doubling in size of the reserve carbohydrate pools. Moreover, stoichiometric measurements indicated that degradation of both chloroplastic reserves and soluble sugars were necessary to supply phosphoenolpyruvate for the synthesis of organic acids at night. Results also suggest that two pools of soluble sugars exist in leaves of C. minor that perform CAM, one a vacuolar pool enriched in 13C and the second a transport pool depleted in 13C. Estimates of carbon-isotope discrimination expressed during CAM, derived from the trafficking among inorganic carbon, organic acids, and carbohydrate pools overnight, ranged from 0.9 to 3.1%[per mille (thousand) sign]. The [delta]13C of structural material did not change significantly between wet and dry seasons, indicating that most of the carbon used in growth was derived from C3 carboxylation.

  4. Hetero-organic thymus antigens.

    PubMed

    Beletskaya, L V; Gnezditskaya, E V

    1985-01-01

    The use of sera containing antibodies to tissue-specific antigens of highly specialized organs (skeletal muscles, heart, skin, excretory glands) enabled us to detect, by immunofluorescence, cells capable of synthesizing analogous antigens (i.e. hetero-organic thymus antigens) in human and animal thymus. Detection of hetero-organic antigens in the thymus is the basis for the hypothesis that natural immunological tolerance to tissue self antigens is formed within the thymus in the course of T-lymphocyte maturation, with thymus antigens taking part in the process.

  5. Bombyx mori nucleopolyhedrovirus nucleic acid binding proteins BRO-B and BRO-E associate with host T-cell intracellular antigen 1 homologue BmTRN-1 to influence protein synthesis during infection.

    PubMed

    Kotani, Eiji; Muto, Sayaka; Ijiri, Hiroshi; Mori, Hajime

    2015-07-01

    Previous reports have indicated that the Bombyx mori nucleopolyhedrovirus (BmNPV) nucleic acid binding proteins BRO-B and BRO-E are expressed during the early stage of infection and that the BRO family likely supports the regulation of mRNA; however, no study has directly examined the function of BRO family proteins in virus-permissive cells. Here, we show that BRO-B and BRO-E associate with cellular T-cell intracellular antigen 1 homologue (BmTRN-1), a translational regulator, and other cellular translation-related proteins in silkworm cells during viral infection. We created BM-N cells that expressed BRO-B/E to study molecular interactions between BmTRN-1 and BRO-B/E and how they influenced protein synthesis. Fluorescent microscopy revealed that BmTRN-1 was localized in cytoplasmic foci during BmNPV infection. Immunofluorescence studies confirmed that BmTRN-1 and BRO-B/E were colocalized in the amorphous conspicuous cytoplasmic foci. Reporter gene studies revealed that co-expression of BRO-B/E synergistically led to a significant decrease in protein synthesis from a designed transcript carrying the 5'untranslated region of a cellular mRNA with no significant change of transcript abundance. Additionally, RNA interference-mediated knockdown of BmTRN-1 resulted in a marked inhibition of the ability of BRO-B/E to regulate the transcript. These results suggested that the association of BmTRN-1 with BRO-B/E is responsible for the inhibitory regulation of certain mRNAs at the post-transcriptional level and add an additional mechanism for how baculoviruses control protein synthesis during infection.

  6. Fractionation of the genetic variants of human alpha 1-acid glycoprotein in the native form by chromatography on an immobilized copper(II) affinity adsorbent. Heterogeneity of the separate variants by isoelectrofocusing and by concanavalin A affinity chromatography.

    PubMed

    Hervé, F; Gomas, E; Duché, J C; Tillement, J P

    1993-05-19

    Fractionation of the three main genetic variants (F1, S and A) of human alpha 1-acid glycoprotein (AAG), in their native (sialylated) form, by chromatography on immobilized copper(II) affinity adsorbent was investigated. This chromatographic method had been previously developed to fractionate the desialylated protein variants. For that purpose, the three main AAG phenotypes samples (F1S/A, F1/A and S/A), which had been previously isolated from individual human plasma samples, and an AAG sample from commercial source (a mixture of the phenotypes) were used in the native form. Affinity chromatography of these different samples on an iminodiacetate Sepharose-copper(II) gel at pH 7 resolved two protein peaks, irrespective of the origin of the native AAG sample used. The unbound peak 1 was found to consist of the F1, the S or both variants, depending on the phenotype of the AAG sample used in the chromatography. The bound peak 2 was found to consist of the A variant in a pure form. The fractionation results obtained with native AAG were found to be the same as those originally yielded by the desialylated protein. However, comparison of the interactions of native and desialylated AAG with immobilized copper(II) ions, using an affinity chromatographic method and a non-chromatographic equilibrium binding technique, respectively, showed that desialylation increased the non-specific interactions of the protein with immobilized copper(II) ions. The AAG variants were not fractionated when affinity chromatography was performed using immobilized zinc, nickel or cobalt(II) ions, instead of copper. After purification of each variant in the sialylated form (F1, S and A), their respective heterogeneity was studied by analytical isoelectrofocusing with carrier ampholytes in the pH range 2.5-4.5. In addition, the lectin-binding behaviour of the separate sialylated AAG variants was investigated by affinity chromatography on immobilized concanavalin A.

  7. Antigenic modulation of metastatic breast and ovary carcinoma cells by intracavitary injection of IFN-alpha.

    PubMed Central

    Giacomini, P.; Mottolese, M.; Fraioli, R.; Benevolo, M.; Venturo, I.; Natali, P. G.

    1992-01-01

    Antigenic modulation of major histocompatibility and tumour associated antigens was observed in neoplastic cells obtained from patients with pleural and abdominal effusions of breast and ovary carcinomas following a single intracavitary dose of 18 x 10(6) U recombinant IFN-alpha. This regimen resulted in antigenic modulation in seven out of 11 tested cases, suggesting a potential, although limited, responsiveness of at least a fraction of breast and ovary carcinoma cells to in situ biomodification with IFN-alpha. PMID:1503908

  8. Human liver nucleolar antigens.

    PubMed

    Busch, R K; Busch, H

    1981-10-01

    In an extension of previous studies on the antigens in rat liver nucleoli (R. K. Busch, R. C. Reddy, D. H. Henning, and H. Busch, Proc. Soc. Exp. Biol. Med. 160, 185 (1979); R. K. Busch and H. Busch, Tumori 63, 347 (1977); F. M. Davis, R. K. Busch, L. C. Yeoman, and H. Busch, Cancer Res. 38, 1906 (1978), rabbit antibodies were elicited to human liver nucleoli isolated by the sucrose--Mg2+ method (10). Fluorescent nucleoli were found in liver cryostat sections treated with rabbit anti-human liver nucleolar antibodies followed by fluorescein-conjugated goat anti-rabbit antibodies. In HeLa cells, fluorescence was distributed throughout the nucleus and in a nuclear network but was not localized to the nucleolus. In placental cryostat sections, an overall nuclear fluorescence was observed with some localization to nucleoli. Immunodiffusion analysis revealed two immunoprecipitin bands which appeared to be liver specific. Other immunoprecipitin bands were common to liver, placenta, and HeLa nuclear extracts. Rocket immunoelectrophoresis revealed two liver-specific antigens, one migrating to the cathode and the other to the anode Other rockets exhibited identity to antigens of other nuclear extracts. These results demonstrate the presence of human liver nucleolar-specific antigens which were not found in the HeLa and placental cells.

  9. Antigen smuggling in tuberculosis.

    PubMed

    Hudrisier, Denis; Neyrolles, Olivier

    2014-06-11

    The importance of CD4 T lymphocytes in immunity to M. tuberculosis is well established; however, how dendritic cells activate T cells in vivo remains obscure. In this issue of Cell Host & Microbe, Srivastava and Ernst (2014) report a mechanism of antigen transfer for efficient activation of antimycobacterial T cells.

  10. Novel integrated mechanical biological chemical treatment (MBCT) systems for the production of levulinic acid from fraction of municipal solid waste: A comprehensive techno-economic analysis.

    PubMed

    Sadhukhan, Jhuma; Ng, Kok Siew; Martinez-Hernandez, Elias

    2016-09-01

    This paper, for the first time, reports integrated conceptual MBCT/biorefinery systems for unlocking the value of organics in municipal solid waste (MSW) through the production of levulinic acid (LA by 5wt%) that increases the economic margin by 110-150%. After mechanical separation recovering recyclables, metals (iron, aluminium, copper) and refuse derived fuel (RDF), lignocelluloses from remaining MSW are extracted by supercritical-water for chemical valorisation, comprising hydrolysis in 2wt% dilute H2SO4 catalyst producing LA, furfural, formic acid (FA), via C5/C6 sugar extraction, in plug flow (210-230°C, 25bar, 12s) and continuous stirred tank (195-215°C, 14bar, 20min) reactors; char separation and LA extraction/purification by methyl isobutyl ketone solvent; acid/solvent and by-product recovery. The by-product and pulping effluents are anaerobically digested into biogas and fertiliser. Produced biogas (6.4MWh/t), RDF (5.4MWh/t), char (4.5MWh/t) are combusted, heat recovered into steam generation in boiler (efficiency: 80%); on-site heat/steam demand is met; balance of steam is expanded into electricity in steam turbines (efficiency: 35%).

  11. Fermentation and dry fractionation increase bioactivity of cloudberry (Rubus chamaemorus).

    PubMed

    Puupponen-Pimiä, Riitta; Nohynek, Liisa; Juvonen, Riikka; Kössö, Tuija; Truchado, Pilar; Westerlund-Wikström, Benita; Leppänen, Tiina; Moilanen, Eeva; Oksman-Caldentey, Kirsi-Marja

    2016-04-15

    Phenolic composition and bioactivity of cloudberry was modified by bioprocessing, and highly bioactive fractions were produced by dry fractionation of the press cake. During fermentation polymeric ellagitannins were partly degraded into ellagic acid derivatives. Phenolic compounds were differentially distributed in seed coarse and fine fractions after dry fractionation process. Tannins concentrated in fine fraction, and flavonol derivatives were mainly found in coarse fraction. Ellagic acid derivatives were equally distributed between the dry fractions. Fermentation and dry fractionation increased statistically significantly anti-adhesion and anti-inflammatory activity of cloudberry. The seed fine fraction showed significant inhibition of P fimbria-mediated haemagglutination assay of uropathogenic Escherichia coli. The seed coarse fraction significantly reduced NO and IL-6 production and iNOS expression in activated macrophages. Fermentation did not affect antimicrobial activity, but slight increase in activity was detected in dry fractions. The results indicate the potential of cloudberry in pharma or health food applications.

  12. Contamination of passenger trains with Dermatophagoides (Acari: Pyroglyphidae) mite antigen in Japan.

    PubMed

    Uehara, K; Toyoda, Y; Konishi, E

    2000-01-01

    Passenger trains were surveyed for contamination with Dermatophagoides farinae Hughes and Dermatophagoides pteronyssinus (Trouesart) mites in Japan. A total of 492 dust samples were collected from upholstered seats in six commuter trains, one long-distance express train and three night trains in October, 1996 and January, April, and July, 1997. Mite antigen levels contained in fine dust fractions of these samples were measured by an enzyme-linked immunosorbent assay. Most samples obtained from commuter trains showed relatively high mite antigen levels of > 10 microgm(-2) (corresponding to > 100 mites). Express and night trains showed lower antigen levels per square meter, but higher mite antigen levels per gram of fine dust than commuter trains, indicating relatively high mite antigen densities. Seasonal comparisons indicated that commuter trains showed the highest mean antigen level per square meter in winter (January), whereas the highest antigen level per gram of fine dust was observed in summer (July) in express and night trains.

  13. HLA-DR antigens in systemic lupus erythematosus: association with specificity of autoantibody responses to nuclear antigens.

    PubMed Central

    Smolen, J S; Klippel, J H; Penner, E; Reichlin, M; Steinberg, A D; Chused, T M; Scherak, O; Graninger, W; Hartter, E; Zielinski, C C

    1987-01-01

    HLA-DR antigens and autoantibodies to the nuclear or cytoplasmic antigens Ro/SSA, La/SSB, Sm, and RNP were determined in North American and Austrian patients with systemic lupus erythematosus (SLE). Analysis of the association of antibodies to these ribonucleic acid (RNA)-protein antigens with HLA-DR antigens showed that HLA-DR3 was related to the presence of anti-Ro/SSA or anti-La/SSB, or both. In contrast, anti-Sm or anti-RNP, or both were associated with HLA-DR4. HLA-DR5 was associated with absence of these autoantibodies. The data extend evidence for the complexity and heterogeneity of SLE. Moreover, they indicate that, in SLE, genes linked to those coding for HLA-DR antigens, are related to the specificity of autoantibody responses rather than to the primary immunological abnormalities of this disorder. PMID:3498447

  14. Flow field-flow fractionation-inductively coupled optical emission spectrometric investigation of the size-based distribution of iron complexed to phytic and tannic acids in a food suspension: implications for iron availability.

    PubMed

    Purawatt, Sopon; Siripinyanond, Atitaya; Shiowatana, Juwadee

    2007-10-01

    Flow field-flow fractionation-inductively coupled plasma optical emission spectrometry (FlFFF-ICP-OES) was applied to achieve the size-based fractionation of iron in a food suspension in order to gain insights into iron availability. The binding of iron with phytic and tannic acids, employed as model inhibitors of iron availability in foods, was investigated at pH 2.0 (representing stomach fluid), pH 5.0 (the transition stage in the upper part of the duodenum), and pH 7.0 (the small intestine). In the presence of phytic acid, iron was found as a free ion or it was associated with molecules smaller than 1 kDa at pH 2.0. Iron associated with molecules larger than 1 kDa when the pH of the mixture was raised to 5.0 and 7.0. In the presence of tannic acid, iron was again mostly associated with molecules smaller than 1 kDa at pH 2.0. However, at pH 5.0, iron and tannic acid associated in large molecules (approximately 25 kDa), while at pH 7.0, most of the iron was associated with macromolecules larger than 500 kDa. Iron size-based distributions of kale extract and tea infusion containing phytic and tannic acids, respectively, were also examined at the three pH values, with and without enzymatic digestion. Without enzymatic digestion of the kale extract and the tea infusion at pH 2.0, most of the iron was released as free ions or associated with molecules smaller than 1 kDa. At other pH values, most of the iron in the kale extract and the tea infusion was found to bind with ~2 kDa and >500 kDa macromolecules, respectively. Upon enzymatic gastrointestinal digestion, the iron was not observed to bind to macromolecules >1 kDa but <500 kDa, due to the enzymatic breakdown of large molecules to smaller ones (<1 kDa).

  15. Standardization and characterization of antigens for the diagnosis of aspergillosis.

    PubMed

    Stopiglia, Cheila Denise Ottonelli; Arechavala, Alicia; Carissimi, Mariana; Sorrentino, Julia Medeiros; Aquino, Valério Rodrigues; Daboit, Tatiane Caroline; Kammler, Luana; Negroni, Ricardo; Scroferneker, Maria Lúcia

    2012-04-01

    The aim of this study was to develop and characterize antigens for the diagnosis of aspergillosis. Nine strains of Aspergillus species Aspergillus fumigatus , Aspergillus flavus , and Aspergillus niger were grown in Sabouraud and Smith broth to produce exoantigens. The antigens were tested by immunodiffusion against sera from patients with aspergillosis and other systemic mycoses. The protein fraction of the antigens was detected by SDS-PAGE; Western blot and representative bands were assessed by mass spectrometry coupled to a nano Acquity UltraPerformance LC and analyzed by the Mascot search engine. Concurrently, all sera were tested with Platelia Aspergillus EIA. The most reactive antigens to sera from patients infected by A. fumigatus were produced by A. fumigatus MG2 Sabouraud and pooled A. fumigatus Sabouraud samples, both with a sensitivity of 93% and specificity of 100% and 97%, respectively. Aspergillus niger and A. flavus antigens were reactive against A. niger and A. flavus sera, each one with a sensitivity and specificity of 100%. Two proteins, probably responsible for antigenic activity, β-glucosidase in A. fumigatus and α-amylase in A. niger were attained. The commercial kit had a specificity of 22%, sensitivity of 100%, positive predictive value of 48%, and negative predictive value of 100%. The antigens produced showed high sensitivity and specificity and can be exploited for diagnostics of aspergilloma.

  16. Stereospecific dehydrogenation of (25R)- and (25S)-3 alpha,7 alpha,12 alpha-trihydroxy-5 beta-cholestanoic acids by acyl-CoA oxidase in rat liver light mitochondrial fraction.

    PubMed

    Ikegawa, S; Watanabe, H; Goto, T; Mano, N; Goto, J; Nambara, T

    1995-08-01

    From a stereochemical point of view, the dehydrogenation mechanism of the biotransformation of 3 alpha,7 alpha,12 alpha-trihydroxy-5 beta-cholestanoic acid (THCA) into (24E)-3 alpha,7 alpha,12 alpha-trihydroxy-5 beta-cholest-24-enoic acid (delta 24-THCA) has been studied with capillary gas chromatography (GC)/negative ion chemical ionization (NICI)-mass spectrometry. After incubation of (24R,25R)- or (24S,25S)-[24,25-2H2]THCA, synthesized from (24E)-delta 24-THCA by a deuterated diimide reduction, with a rat liver light mitochondrial fraction, 5 beta-cholestanoic acids were extracted and derivatized into a pentafluorobenzyl (PFB) ester-dimethylethylsilyl (DMES) ether. Subsequent resolution into THCA and delta 24-THCA was attained by GC on a cross-linked 5% phenylmethyl silicone fused-silica capillary column monitored with a corresponding characteristic carboxylate anion [M-PFB]- in the NICI mode. The stereospecific elimination of a pro-R hydrogen at C-24 in both (25R)- and (25S)-THCA indicated syn-elimination for the former, whereas anti-elimination for the latter was observed.

  17. Characteristics of Immunochemical Interrelations between Cellular Components Included into the Composition of Allergenoactive Fractions of Enterobacteria of Various Genera,

    DTIC Science & Technology

    Cellular components which form allergenoactive fractions include antigens common to enterobacteria of different genera. In Escherichia, Shigella...The degree of immunochemical community is expressed identically for individual examples of enterobacteria . According to the number of common antigens...in the allergenic fractions, enterobacteria can be divided into four immunochemical types: Escherichia (combines the genera Escherichia, Salmonella

  18. [Analysis on the impact of composting with different proportions of corn stalks and pig manure on humic acid fractions and IR spectral feature].

    PubMed

    Sun, Xiang-Ping; Li, Guo-Xue; Xiao, Ai-Ping; Shi, Hong; Wang, Yi-Ming; Li, Yang-Yang

    2014-09-01

    Using pig manure and corn straw as raw materials for high-temperature composting, setting three different treat- ments: C/N 15, C/N 25, and C/N 35. Composting period is 120 days, which contains 30 days for ventilation cycle by forced continuous ventilation. Sampled on 0, 22, 30, 60, 90, 120th days, they were analyzed by elemental analysis and IR spectroscopy to study effect of different lignin content on compost humic acid (HA) composition and molecular structure. The results showed that the change in composting humic acid C focused on the first 30 days, while after composting, the O/C of compost HA increased, H/C decreased, and N content increased. Low C/N (15) and higher C/N ratio (35) had higher degree of oxidation than the C/N 25 in compost HA. FTIR indicated that the infrared spectrum shapes with different lignin content treatment are similar during the composting process, but the peak intensity is obviously different. Research results proved that the composting stage is more conducive to enhanced aromatic in compost HA. After composting, C/N 15 had less polysaccharide and fat ingredients and more aromatic structural components in compost HA, compared with C/N 25 and 35. In addition, compost HA of C/N 15 had higher degree of humification and its structure was more stable.

  19. Fractional vector calculus and fractional Maxwell's equations

    SciTech Connect

    Tarasov, Vasily E.

    2008-11-15

    The theory of derivatives and integrals of non-integer order goes back to Leibniz, Liouville, Grunwald, Letnikov and Riemann. The history of fractional vector calculus (FVC) has only 10 years. The main approaches to formulate a FVC, which are used in the physics during the past few years, will be briefly described in this paper. We solve some problems of consistent formulations of FVC by using a fractional generalization of the Fundamental Theorem of Calculus. We define the differential and integral vector operations. The fractional Green's, Stokes' and Gauss's theorems are formulated. The proofs of these theorems are realized for simplest regions. A fractional generalization of exterior differential calculus of differential forms is discussed. Fractional nonlocal Maxwell's equations and the corresponding fractional wave equations are considered.

  20. Purification, characterization, and localization of a protein antigen shared by thermophilic campylobacters.

    PubMed Central

    Dubreuil, J D; Kostrzynska, M; Logan, S M; Harris, L A; Austin, J W; Trust, T J

    1990-01-01

    A protein antigen with an apparent molecular weight (Mr) of 31,000 was isolated from 0.2 M glycine hydrochloride (pH 2.2) extracts of a typical human fecal isolate, Campylobacter jejuni VC74. The protein was purified to homogeneity on a preparative scale by immunoaffinity chromatography followed by molecular sieving with a Superose 12 column. Isoelectric focusing under nondenaturing conditions indicated a pI of 9.3, and amino acid composition analysis showed that the protein was unusually rich in lysine, containing 14.9 mol% of this basic amino acid. Cysteine and tryptophan were absent. The protein also contained approximately 35% hydrophobic amino acid residues, and N-terminal amino acid analysis showed that 17 of the first 38 residues were hydrophobic. This amino-terminal sequence to residue 22 was virtually identical to that of an antigenically cross-reactive 31,000-Mr protein isolated from another C. jejuni strain belonging to a different heat-labile serogroup. Western blotting (immunoblotting) of glycine extracts of other C. jejuni, Campylobacter coli, and Campylobacter laridis strains belonging to different thermolabile and thermostable serotypes, as well as Campylobacter fetus, with a rabbit polyclonal antiserum raised against the purified C. jejuni VC74 protein showed that all C. jejuni, C. coli, and C. laridis strains tested contained a 31,000-Mr protein with epitopes which were antigenically cross-reactive with the C. jejuni VC74 protein. The antigenically cross-reactive epitopes of this protein were also readily detected by immunodot blot assay of glycine extracts of C. jejuni, C. coli, and C. laridis with monospecific polyclonal antisera to the 31,000-Mr protein, suggesting that this serological test could be a useful addition to those currently employed in the rapid identification of these important pathogens. Slide agglutination reactions, immunofluorescence assay, and immunogold electron microscopy with antisera to purified 31,000-Mr protein and

  1. Evaluation of the therapeutic effect of percutaneous nephroureterolithotomy by Tc-99m diethylenetiaminepentaacetic acid (DTPA) renal scintigraphy--alteration of the renal fraction of blood flow, split-GFR, and renal mean transit time.

    PubMed

    Ishibashi, M; Morita, S; Rabito, C A; Umezaki, N; Matsuoka, K; Noda, S; Eto, K; Ohtake, H

    1990-01-01

    To evaluate the therapeutic effects of percutaneous nephroureterolithotomy, the renal function of eleven patients with renal calculi was studied, pre- and post-intervention. Renal function was determined, by renal scintigraphy with the renal agent, Tc-99m diethylenetriaminepentaacetic acid (DTPA). In each renal scintigram the renogram curve was analyzed and the following were determined by deconvolution analysis; the renal fraction of blood flow (RFBF), DTPA-glomerular filtration ratio (GFR), and the renal mean transit time (MTT). The successful results in percutaneous nephroureterolithotomy (PNL) was proven using the radionuclide technique in most cases. From these results it can be concluded that renal scintigraphy is an effective procedure to evaluate the effect of PNL for treating renal calculi and secondary hydronephrosis.

  2. Antibodies to staphylococcal peptidoglycan and its peptide epitopes, teichoic acid, and lipoteichoic acid in sera from blood donors and patients with staphylococcal infections.

    PubMed Central

    Wergeland, H I; Haaheim, L R; Natås, O B; Wesenberg, F; Oeding, P

    1989-01-01

    Antibodies to the staphylococcal antigens peptidoglycan, beta-ribitol teichoic acid, and lipoteichoic acid, as well as to the peptidoglycan epitopes L-Lys-D-Ala-D-Ala, L-Lys-D-Ala, and pentaglycine, were found over a wide range of concentrations in sera from both blood donors and patients with verified or suspected staphylococcal infections. The patient group was heterogeneous with regard to both age and type of staphylococcal infections, being representative for sera sent to our laboratory. In single-antigen assays antibodies to pentaglycine had the highest predictive positive value (67%), although only 32% of the patients had elevated levels of such antibodies. Combinations of test antigens could yield positive predictive values as high as 100%, but then the fraction of positive sera was low. Indeed, the fraction of patient sera which was positive in multiple-antigen tests never exceeded 61%. The clinical usefulness of these seroassays for identifying Staphylococcus aureus as a causative agent was limited, owing to the considerable overlap in the range of antibody concentrations between patient and blood donor sera. PMID:2473994

  3. Carbohydrate-Mediated Targeting of Antigen to Dendritic Cells Leads to Enhanced Presentation of Antigen to T Cells

    PubMed Central

    Adams, Eddie W.; Ratner, Daniel M.; Seeberger, Peter H.; Hacohen, Nir

    2009-01-01

    The unique therapeutic value of dendritic cells (DCs) for the treatment of allergy, autoimmunity and transplant rejection is predicated upon our ability to selectively deliver antigens, drugs or nucleic acids to DCs in vivo. Here we describe a method for delivering whole protein antigens to DCs based on carbohydrate-mediated targeting of DC-expressed lectins. A series of synthetic carbohydrates was chemically-coupled to a model antigen, ovalbumin (OVA), and each conjugate was evaluated for its ability to increase the efficiency of antigen presentation by murine DCs to OVA-specific T cells (CD4+ and CD8+). In vitro data are presented that demonstrate that carbohydrate modification of OVA leads to a 50-fold enhancement of presentation of antigenic peptide to CD4+ T cells. A tenfold enhancement is observed for CD8+ T cells; this indicates that the targeted lectin(s) can mediate cross-presentation of antigens on MHC class I. Our data indicate that the observed enhancements in antigen presentation are unique to OVA that is conjugated to complex oligosaccharides, such as a high-mannose nonasaccharide, but not to monosaccharides. Taken together, our data suggest that a DC targeting strategy that is based upon carbohydrate-lectin interactions is a promising approach for enhancing antigen presentation via class I and class II molecules. PMID:18186095

  4. Antioxidant activity of cod (Gadus morhua) protein hydrolysates: Fractionation and characterisation of peptide fractions.

    PubMed

    Sabeena Farvin, K H; Andersen, Lisa Lystbæk; Otte, Jeanette; Nielsen, Henrik Hauch; Jessen, Flemming; Jacobsen, Charlotte

    2016-08-01

    This study aimed to characterise peptide fractions (>5kDa, 3-5kDa and <3kDa) with antioxidative activity obtained from a cod protein hydrolysate. The free amino acids in all fractions were dominated by Ala, Gly, Glu and Ser. The total amino acid composition had high proportions of Lys, Ala and Glu. The 3-5kDa and <3kDa fractions were further fractionated by size exclusion chromatography. All sub-fractions showed high Fe(2+) chelating activity. The DPPH radical-scavenging activity of the 3-5kDa fraction was exerted mainly by one sub-fraction dominated by peptides with masses below 600Da. The DPPH radical-scavenging activity of the <3kDa fraction was exerted by sub-fractions with low molecular weight. The highest reducing power was found in a sub-fraction containing peptides rich in Arg, Tyr and Phe. Both free amino acids and low molecular weight peptides thus seemed to contribute to the antioxidative activity of the peptide fractions, and Tyr seemed to play a major role in the antioxidant activity.

  5. Comparison of anti-inflammatory activities of an anthocyanin-rich fraction from Portuguese blueberries (Vaccinium corymbosum L.) and 5-aminosalicylic acid in a TNBS-induced colitis rat model

    PubMed Central

    Pereira, Sónia R.; Pereira, Rita; Figueiredo, Isabel; Freitas, Victor; Dinis, Teresa C. P.; Almeida, Leonor M.

    2017-01-01

    Despite the actual therapeutic approaches for inflammatory bowel disease (IBD), efficient and secure alternative options remain a research focus. In this context, anthocyanins seem promising natural anti-inflammatory agents, but their action mechanisms and efficacy as compared with established drugs still require more clarification. The main aim of this study was to compare the anti-inflammatory action of a chemically characterized anthocyanin-rich fraction (ARF), obtained from Portuguese blueberries (Vaccinium corymbosum L.), with that of 5-aminosalicylic acid (5-ASA), a first-line drug in IBD, in a 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced colitis rat model. Such fraction showed a high content and great molecular diversity of anthocyanins, with malvidin-3-galactoside and petunidin-3-arabinoside in the highest concentrations. After daily administration by intragastric infusion for 8 days, ARF, at a molar anthocyanin concentration about 30 times lower than 5-ASA, showed a higher effectiveness in counteracting the intestinal inflammation, as assessed by i) body weight variation and colon damage score, ii) reduction in leukocyte infiltration, iii) increase in antioxidant defenses and iv) by downregulation of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in colon tissue homogenates. The strong inhibition of COX-2 expression seems to be a crucial anti-inflammatory mechanism common to both ARF and 5-ASA, but the additional higher abilities of anthocyanins to downregulate iNOS and to decrease leukocytes infiltration and to increase antioxidant defenses in colon may account for the much higher anti-inflammatory action of anthocyanins. These data may contribute to the development of a promising natural approach in IBD management. PMID:28329021

  6. Initialized Fractional Calculus

    NASA Technical Reports Server (NTRS)

    Lorenzo, Carl F.; Hartley, Tom T.

    2000-01-01

    This paper demonstrates the need for a nonconstant initialization for the fractional calculus and establishes a basic definition set for the initialized fractional differintegral. This definition set allows the formalization of an initialized fractional calculus. Two basis calculi are considered; the Riemann-Liouville and the Grunwald fractional calculi. Two forms of initialization, terminal and side are developed.

  7. Tempered fractional calculus

    SciTech Connect

    Sabzikar, Farzad; Meerschaert, Mark M.; Chen, Jinghua

    2015-07-15

    Fractional derivatives and integrals are convolutions with a power law. Multiplying by an exponential factor leads to tempered fractional derivatives and integrals. Tempered fractional diffusion equations, where the usual second derivative in space is replaced by a tempered fractional derivative, govern the limits of random walk models with an exponentially tempered power law jump distribution. The limiting tempered stable probability densities exhibit semi-heavy tails, which are commonly observed in finance. Tempered power law waiting times lead to tempered fractional time derivatives, which have proven useful in geophysics. The tempered fractional derivative or integral of a Brownian motion, called a tempered fractional Brownian motion, can exhibit semi-long range dependence. The increments of this process, called tempered fractional Gaussian noise, provide a useful new stochastic model for wind speed data. A tempered fractional difference forms the basis for numerical methods to solve tempered fractional diffusion equations, and it also provides a useful new correlation model in time series.

  8. Chiroptical characterization of homopolymeric block fractions in alginates.

    PubMed

    Martínez-Gómez, Fabián; Mansilla, Andrés; Matsuhiro, Betty; Matulewicz, María C; Troncoso-Valenzuela, Marcos A

    2016-08-01

    Homopolymannuronic and homopolyguluronic fractions were obtained by partial hydrolysis of the alkaline extracts from the brown seaweeds Ascoseira mirabilis, Desmarestia menziessi, Desmarestia ligulata and Durvillaea sp. collected in southern Chile. Full characterization of the fractions was achieved by FT-IR and NMR spectroscopy. Total hydrolysis with 90% formic acid of the homopolymeric fractions allowed the preparation of mannuronic and guluronic acids. Both monomers and homopolymeric fractions as neutral salts were studied by CD and ORD. Chiroptical spectra were similar in shape and sign to those previously published in the literature, and permitted to assign D configuration to mannuronic acid and L configuration to guluronic acid in alginic acids. Specific optical rotation values at the sodium D light for the homopolymannuronic (∼-100°) and homopolyguluronic (∼-110°) acid fractions were obtained. These high negative values are proposed for the assignment of the absolute configuration of monomers in homopolymeric fractions.

  9. Effect of pH on antigen binding by clonotypic antibodies with different isoelectric points

    SciTech Connect

    Endo, Y.; Miyai, K.; Hata, N.; Iijima, Y.

    1987-02-01

    Polyclonal rabbit antibodies to thyroxine, human myoglobin, human growth hormone, human thyrotropin, human alpha-fetoprotein, and human thyroglobulin were fractionated into clonotypic antibodies with different isoelectric points by agarose isoelectric focusing or chromatofocusing. The effect of pH on the binding of these antigens by their respective clonotypic antibodies was assessed by radioimmunoassay. The profiles of the pH effect differed both for different antigens and for different pI's of the antibodies used. The pH optima in the radioimmunoassays for protein antigens were found to be expressed as a function of pI and molecular weight of both antigen and antibody molecules.

  10. Evidence for glycosyl-phosphatidylinositol anchoring of Toxoplasma gondii major surface antigens

    SciTech Connect

    Tomavo, S.; Schwarz, R.T.; Dubremetz, J.F. )

    1989-10-01

    The four major surface antigens of Toxoplasma gondii tachyzoites (P43, P35, P30, and P22) were made water soluble by phosphatidylinositol-specific phospholipase C (PI-PLC). These antigens were biosynthetically labeled with {sup 3}H-fatty acids, ({sup 3}H)ethanolamine, and ({sup 3}H)carbohydrates. Treatment of {sup 3}H-fatty-acid-labeled parasite lysates with PI-PLC removed the radioactive label from these antigens. A cross-reacting determinant was exposed on these antigens after PI-PLC treatment.

  11. Evidence for glycosyl-phosphatidylinositol anchoring of Toxoplasma gondii major surface antigens.

    PubMed Central

    Tomavo, S; Schwarz, R T; Dubremetz, J F

    1989-01-01

    The four major surface antigens of Toxoplasma gondii tachyzoites (P43, P35, P30, and P22) were made water soluble by phosphatidylinositol-specific phospholipase C (PI-PLC). These antigens were biosynthetically labeled with 3H-fatty acids, [3H]ethanolamine, and [3H]carbohydrates. Treatment of 3H-fatty-acid-labeled parasite lysates with PI-PLC removed the radioactive label from these antigens. A cross-reacting determinant was exposed on these antigens after PI-PLC treatment. Images PMID:2531282

  12. Predicted complementarity determining regions of the T cell antigen receptor determine antigen specificity.

    PubMed Central

    Katayama, C D; Eidelman, F J; Duncan, A; Hooshmand, F; Hedrick, S M

    1995-01-01

    The antigen receptor on T cells (TCR) has been predicted to have a structure similar to a membrane-anchored form of an immunoglobulin F(ab) fragment. Virtually all of the conserved amino acids that are important for inter- and intramolecular interactions in the VH-VL pair are also conserved in the TCR V alpha and V beta chains. A molecular model of the TCR has been constructed by homology and we have used the information from this, as well as the earlier structural predictions of others, to study the basis for specificity. Specifically, regions of a TCR cloned from an antigen-specific T cell were stitched into the corresponding framework of a second TCR. Results indicate that the substitution of amino acid sequences corresponding to the complementarity determining regions (CDRs) of immunoglobulin can convey the specificity for antigen and major histocompatibility complex molecules. These data are consistent with a role, but not an exclusive role, for CDR3 in antigen peptide recognition. Images PMID:7534228

  13. Fraction Sense: Foundational Understandings.

    PubMed

    Fennell, Francis Skip; Karp, Karen

    2016-08-09

    The intent of this commentary is to identify elements of fraction sense and note how the research studies provided in this special issue, in related but somewhat different ways, validate the importance of such understandings. Proficiency with fractions serves as a prerequisite for student success in higher level mathematics, as well as serving as a gateway to many occupations and varied contexts beyond the mathematics classroom. Fraction sense is developed through instructional opportunities involving fraction equivalence and magnitude, comparing and ordering fractions, using fraction benchmarks, and computational estimation. Such foundations are then extended to operations involving fractions and decimals and applications involving proportional reasoning. These components of fraction sense are all addressed in the studies provided in this issue, with particular consideration devoted to the significant importance of the use of the number line as a central representational tool for conceptually understanding fraction magnitude.

  14. TEMPERED FRACTIONAL CALCULUS

    PubMed Central

    MEERSCHAERT, MARK M.; SABZIKAR, FARZAD; CHEN, JINGHUA

    2014-01-01

    Fractional derivatives and integrals are convolutions with a power law. Multiplying by an exponential factor leads to tempered fractional derivatives and integrals. Tempered fractional diffusion equations, where the usual second derivative in space is replaced by a tempered fractional derivative, govern the limits of random walk models with an exponentially tempered power law jump distribution. The limiting tempered stable probability densities exhibit semi-heavy tails, which are commonly observed in finance. Tempered power law waiting times lead to tempered fractional time derivatives, which have proven useful in geophysics. The tempered fractional derivative or integral of a Brownian motion, called a tempered fractional Brownian motion, can exhibit semi-long range dependence. The increments of this process, called tempered fractional Gaussian noise, provide a useful new stochastic model for wind speed data. A tempered difference forms the basis for numerical methods to solve tempered fractional diffusion equations, and it also provides a useful new correlation model in time series. PMID:26085690

  15. Methamphetamine inhibits antigen processing, presentation, and phagocytosis.

    PubMed

    Tallóczy, Zsolt; Martinez, Jose; Joset, Danielle; Ray, Yonaton; Gácser, Attila; Toussi, Sima; Mizushima, Noboru; Nosanchuk, Joshua D; Nosanchuk, Josh; Goldstein, Harris; Loike, John; Sulzer, David; Santambrogio, Laura

    2008-02-08

    Methamphetamine (Meth) is abused by over 35 million people worldwide. Chronic Meth abuse may be particularly devastating in individuals who engage in unprotected sex with multiple partners because it is associated with a 2-fold higher risk for obtaining HIV and associated secondary infections. We report the first specific evidence that Meth at pharmacological concentrations exerts a direct immunosuppressive effect on dendritic cells and macrophages. As a weak base, Meth collapses the pH gradient across acidic organelles, including lysosomes and associated autophagic organelles. This in turn inhibits receptor-mediated phagocytosis of antibody-coated particles, MHC class II antigen processing by the endosomal-lysosomal pathway, and antigen presentation to splenic T cells by dendritic cells. More importantly Meth facilitates intracellular replication and inhibits intracellular killing of Candida albicans and Cryptococcus neoformans, two major AIDS-related pathogens. Meth exerts previously unreported direct immunosuppressive effects that contribute to increased risk of infection and exacerbate AIDS pathology.

  16. Functional fractionation of platelets.

    PubMed

    Haver, V M; Gear, A R

    1981-02-01

    Studies of platelet populations suggest that they are heterogeneous in size, age, and metabolic parameters. In an attempt to correlate these parameters with efficiency of aggregation, a new technique, functional fractionation, was developed. Platelet populations are separated by their differential reactivity to aggregating agents. For example, low doses of ADP (0.1 to 0.7 microM) are added to stirred PRP, after which gentle centrifugation is used to remove aggregates from single unreacted platelets. The loose aggregates can be readily dispersed for comparison of the physical or biochemical properties of the reacted versus unreacted platelets. It was found that reactive platelets were larger (6.5 micrometer3) than unreacted platelets (5.51 micrometer3). No significant difference in density existed between the two populations, and no release of [14C]serotonin from prelabeled platelets occurred during functional fractionation. Scanning and transmission electron microscopy confirmed the size difference and revealed that in both populations platelets were structurally intact with a normal discoid shape and no significant difference in organelle content. Human platelets most reactive to ADP were also enriched in glycogen (3.6-fold), ATP (1.6-fold), and ADP (twofold), compared with less reactive cells. These "reactive" cells took up more 51[Cr] and contained 1.9 times more surface sialic acid. In an in vivo aging experiment, rats were injected with 75[Se]methionine. Shortly after labeling (1 day), the most reactive platelets possessed the highest amount of 75[Se]. These results reveal that functionally active platelets, which are also larger, are more active metabolically than less reactive platelets, possess a higher negative surface charge, and may be a younger population.

  17. Immune recognition of protein antigens

    SciTech Connect

    Laver, W.G.; Air, G.M.

    1985-01-01

    This book contains 33 papers. Some of the titles are: Antigenic Structure of Influenze Virus Hemagglutinin; Germ-line and Somatic Diversity in the Antibody Response to the Influenza Virus A/PR/8/34 Hemagglutinin; Recognition of Cloned Influenza A Virus Gene Products by Cytotoxic T Lymphocytes; Antigenic Structure of the Influenza Virus N2 Neuraminidase; and The Molecular and Genetic Basis of Antigenic Variation in Gonococcal Pillin.

  18. Changes in structural and antigenic properties of proteins by radiation

    NASA Astrophysics Data System (ADS)

    Kume, Tamikazu; Matsuda, Tsukasa

    1995-08-01

    Radiation effect on structural and antigenic properties of proteins (0.2% in 0.01 M phosphate buffer, pH 7.4) were investigated using ovalbumin (OVA) and bovine serum albumin (BSA). Aggregation of OVA and BSA was induced by radiation and the molecular mass increased significantly in N 2. Significant changes in surface hydrophobicity and [ θ] 222 nm of CD were also observed by radiation showing the destruction of secondary structure of proteins. Antigenicity of irradiated OVA measured by the method of immunodiffusion was decreased by radiation, and the reactivity to anti-OVA antibody was almost diminished at 8 kGy in N 2 and 4 kGy in O 2, respectively. The reactivity of BSA was diminished at 4 kGy both in N 2 and O 2. Changes in hydrophobicity of OVA did not correspond to the decrease in antigenicity, whereas the changes in [ θ] 222 nm relatively well corresponded to the antigenicity. The SDS-PAGE and immunoblotting analysis showed that radiation at higher doses induced the production of protein aggregates and degraded fragments with reactivity to the specific antibodies. These results suggest that the main part of conformation-dependent antigenic structure (conformational epitope) is easily lost by radiation, but some antigenicity, which is mostly due to the amino acid sequence-dependent antigenic structures (sequential epitopes), remains even at higher dose.

  19. Molecular cloning of Taenia taeniaeformis oncosphere antigen genes.

    PubMed

    Cougle, W G; Lightowlers, M W; Bogh, H O; Rickard, M D; Johnson, K S

    1991-03-01

    Infection of mice with the cestode Taenia taeniaeformis exhibits several important features common to other cestode infections, including the ability to vaccinate with crude antigen mixtures. Partial purification of the protective oncosphere antigens has been reported with a cutout from deoxycholate (DOC) acrylamide gels; this cutout was called fraction II (FII), and comprises approximately 10% of total DOC-soluble oncosphere antigen. Western blots of DOC gels probed with anti-FII antisera revealed a series of 3-5 discrete bands within the FII region. Further fractionation of the FII antigens on DOC gels was impractical due to limitations in supply of oncospheres, so a cDNA library was constructed from 150 ng of oncosphere mRNA and screened with alpha-FII antisera. Two distinct clone families were identified, oncA and oncB. Antibodies affinity-purified on either of two representative members, oncA1 and oncB1, recognised all the FII bands. Individual FII bands excised from a DOC gel resolved into an overlapping series of molecules when re-run on SDS-PAGE, indicating that each FII band consisted of several polypeptides of differing molecular weight. Immunoprecipitates resolved on SDS-PAGE revealed that alpha-FII recognised 3 major oncosphere antigens, of 62, 34 and 25 kDa; antisera against oncB precipitated both the 34- and 25-kDa antigens, whereas alpha-oncA antisera precipitated the 62-kDa antigen. We conclude that oncA and oncB encode the major antigens in the FII complex. The 62-kDa antigen encoded by oncA1 was the only common antigen precipitated by anti-FII and two other antisera raised against different protective extracts, suggesting that it may be a protective component in all three. Southern blot results indicate that oncA and oncB are distinct genes present at low copy number in the genome. Evidence is also presented suggesting that some cestode mRNAs, including oncA, may use variant polyadenylation signals.

  20. Exosome targeting of tumor antigens expressed by cancer vaccines can improve antigen immunogenicity and therapeutic efficacy.

    PubMed

    Rountree, Ryan B; Mandl, Stefanie J; Nachtwey, James M; Dalpozzo, Katie; Do, Lisa; Lombardo, John R; Schoonmaker, Peter L; Brinkmann, Kay; Dirmeier, Ulrike; Laus, Reiner; Delcayre, Alain

    2011-08-01

    MVA-BN-PRO (BN ImmunoTherapeutics) is a candidate immunotherapy product for the treatment of prostate cancer. It encodes 2 tumor-associated antigens, prostate-specific antigen (PSA), and prostatic acid phosphatase (PAP), and is derived from the highly attenuated modified vaccinia Ankara (MVA) virus stock known as MVA-BN. Past work has shown that the immunogenicity of antigens can be improved by targeting their localization to exosomes, which are small, 50- to 100-nm diameter vesicles secreted by most cell types. Exosome targeting is achieved by fusing the antigen to the C1C2 domain of the lactadherin protein. To test whether exosome targeting would improve the immunogenicity of PSA and PAP, 2 additional versions of MVA-BN-PRO were produced, targeting either PSA (MVA-BN-PSA-C1C2) or PAP (MVA-BN-PAP-C1C2) to exosomes, while leaving the second transgene untargeted. Treatment of mice with MVA-BN-PAP-C1C2 led to a striking increase in the immune response against PAP. Anti-PAP antibody titers developed more rapidly and reached levels that were 10- to 100-fold higher than those for mice treated with MVA-BN-PRO. Furthermore, treatment with MVA-BN-PAP-C1C2 increased the frequency of PAP-specific T cells 5-fold compared with mice treated with MVA-BN-PRO. These improvements translated into a greater frequency of tumor rejection in a PAP-expressing solid tumor model. Likewise, treatment with MVA-BN-PSA-C1C2 increased the antigenicity of PSA compared with treatment with MVA-BN-PRO and resulted in a trend of improved antitumor efficacy in a PSA-expressing tumor model. These experiments confirm that targeting antigen localization to exosomes is a viable approach for improving the therapeutic potential of MVA-BN-PRO in humans.

  1. Genetic diversity and antigenicity variation of Babesia bovis merozoite surface antigen-1 (MSA-1) in Thailand.

    PubMed

    Tattiyapong, Muncharee; Sivakumar, Thillaiampalam; Takemae, Hitoshi; Simking, Pacharathon; Jittapalapong, Sathaporn; Igarashi, Ikuo; Yokoyama, Naoaki

    2016-07-01

    Babesia bovis, an intraerythrocytic protozoan parasite, causes severe clinical disease in cattle worldwide. The genetic diversity of parasite antigens often results in different immune profiles in infected animals, hindering efforts to develop immune control methodologies against the B. bovis infection. In this study, we analyzed the genetic diversity of the merozoite surface antigen-1 (msa-1) gene using 162 B. bovis-positive blood DNA samples sourced from cattle populations reared in different geographical regions of Thailand. The identity scores shared among 93 msa-1 gene sequences isolated by PCR amplification were 43.5-100%, and the similarity values among the translated amino acid sequences were 42.8-100%. Of 23 total clades detected in our phylogenetic analysis, Thai msa-1 gene sequences occurred in 18 clades; seven among them were composed of sequences exclusively from Thailand. To investigate differential antigenicity of isolated MSA-1 proteins, we expressed and purified eight recombinant MSA-1 (rMSA-1) proteins, including an rMSA-1 from B. bovis Texas (T2Bo) strain and seven rMSA-1 proteins based on the Thai msa-1 sequences. When these antigens were analyzed in a western blot assay, anti-T2Bo cattle serum strongly reacted with the rMSA-1 from T2Bo, as well as with three other rMSA-1 proteins that shared 54.9-68.4% sequence similarity with T2Bo MSA-1. In contrast, no or weak reactivity was observed for the remaining rMSA-1 proteins, which shared low sequence similarity (35.0-39.7%) with T2Bo MSA-1. While demonstrating the high genetic diversity of the B. bovis msa-1 gene in Thailand, the present findings suggest that the genetic diversity results in antigenicity variations among the MSA-1 antigens of B. bovis in Thailand.

  2. Maximizing Tumor Immunity With Fractionated Radiation

    SciTech Connect

    Schaue, Doerthe; Ratikan, Josephine A.; Iwamoto, Keisuke S.; McBride, William H.

    2012-07-15

    Purpose: Technologic advances have led to increased clinical use of higher-sized fractions of radiation dose and higher total doses. How these modify the pathways involved in tumor cell death, normal tissue response, and signaling to the immune system has been inadequately explored. Here we ask how radiation dose and fraction size affect antitumor immunity, the suppression thereof, and how this might relate to tumor control. Methods and Materials: Mice bearing B16-OVA murine melanoma were treated with up to 15 Gy radiation given in various-size fractions, and tumor growth followed. The tumor-specific immune response in the spleen was assessed by interferon-{gamma} enzyme-linked immunospot (ELISPOT) assay with ovalbumin (OVA) as the surrogate tumor antigen and the contribution of regulatory T cells (Tregs) determined by the proportion of CD4{sup +}CD25{sup hi}Foxp3{sup +} T cells. Results: After single doses, tumor control increased with the size of radiation dose, as did the number of tumor-reactive T cells. This was offset at the highest dose by an increase in Treg representation. Fractionated treatment with medium-size radiation doses of 7.5 Gy/fraction gave the best tumor control and tumor immunity while maintaining low Treg numbers. Conclusions: Radiation can be an immune adjuvant, but the response varies with the size of dose per fraction. The ultimate challenge is to optimally integrate cancer immunotherapy into radiation therapy.

  3. Hepatitis C Virus Antigenic Convergence

    PubMed Central

    Campo, David S.; Dimitrova, Zoya; Yokosawa, Jonny; Hoang, Duc; Perez, Nestor O.; Ramachandran, Sumathi; Khudyakov, Yury

    2012-01-01

    Vaccine development against hepatitis C virus (HCV) is hindered by poor understanding of factors defining cross-immunoreactivity among heterogeneous epitopes. Using synthetic peptides and mouse immunization as a model, we conducted a quantitative analysis of cross-immunoreactivity among variants of the HCV hypervariable region 1 (HVR1). Analysis of 26,883 immunological reactions among pairs of peptides showed that the distribution of cross-immunoreactivity among HVR1 variants was skewed, with antibodies against a few variants reacting with all tested peptides. The HVR1 cross-immunoreactivity was accurately modeled based on amino acid sequence alone. The tested peptides were mapped in the HVR1 sequence space, which was visualized as a network of 11,319 sequences. The HVR1 variants with a greater network centrality showed a broader cross-immunoreactivity. The entire sequence space is explored by each HCV genotype and subtype. These findings indicate that HVR1 antigenic diversity is extensively convergent and effectively limited, suggesting significant implications for vaccine development. PMID:22355779

  4. DIY Fraction Pack.

    ERIC Educational Resources Information Center

    Graham, Alan; Graham, Louise

    2003-01-01

    Describes a very successful attempt to teach fractions to year 5 pupils based on pupils making their own fraction pack. Children decided for themselves how to make the fractional slices used in the activity using colored cardboard sheets and templates of a paper circle consisting of 24 equal slices. (Author/NB)

  5. Purification and partial characterization of an antigen specific to Lactobacillus brevis strains with beer spoilage activity.

    PubMed

    Yasui, T; Yoda, K

    1997-06-15

    Certain Lactobacillus brevis strains are resistant to hop-derived compounds such as isohumulone and are able to grow in beer. In this study, we raised an antiserum against our beer spoilage laboratory strain L. brevis 578 which reacted with 23 of 24 beer spoilers and two of 13 non-spoilers in precipitation reactions using 0.5 M NaOH cell extracts. This specific antigen to the beer spoilage L. brevis strains (SABSL) was demonstrated to be located beneath the S-layer proteins by agglutination reactions using S-layer protein-stripped cells obtained by treatment with 0.1 M NaOH. SABSL was purified using an affinity column coupled with an antibody against SABSL. The purified antigen was hydrolyzed with 2 M HCl and the hydrolyzate was analyzed by thin-layer chromatography and enzymatic analysis. The results showed that SABSL contains glycerol, phosphate, glycerophosphate, D-galactose and D-glucose. D-Galactose and D-glucose accounted for 4.7% and 0.1% of the composition, respectively. Melibiose, but not mannose, inhibited the precipitation reaction. Intense precipitation reactions were obtained with fractions which did not bind to the ConA-column. These results indicate that the immunodominant component of the SABSL is galactose and the SABSL determinant is most probably a galactosylated glycerol teichoic acid. The antiserum raised against the beer spoilage strain L. brevis 578 could distinguish between Pediococcus beer spoilers and non-spoilers in precipitation reactions.

  6. Allergen Fractions of Bacteria of the Intestinal Family,

    DTIC Science & Technology

    The extracellular fraction obtained by the Ando - Verzhikovskiy method from the bacterial species Enterobacteria cloacae possessed the activity of an...revealed immunoelectrophoretically. Allergens from different strains of Enterobacteria cloacae had a close antigenic structure and caused cross allergic reactions in experiments on sensitized animals.

  7. Novel antigen delivery systems

    PubMed Central

    Trovato, Maria; Berardinis, Piergiuseppe De

    2015-01-01

    Vaccines represent the most relevant contribution of immunology to human health. However, despite the remarkable success achieved in the past years, many vaccines are still missing in order to fight important human pathologies and to prevent emerging and re-emerging diseases. For these pathogens the known strategies for making vaccines have been unsuccessful and thus, new avenues should be investigated to overcome the failure of clinical trials and other important issues including safety concerns related to live vaccines or viral vectors, the weak immunogenicity of subunit vaccines and side effects associated with the use of adjuvants. A major hurdle of developing successful and effective vaccines is to design antigen delivery systems in such a way that optimizes antigen presentation and induces broad protective immune responses. Recent advances in vector delivery technologies, immunology, vaccinology and system biology, have led to a deeper understanding of the molecular and cellular mechanisms by which vaccines should stimulate both arms of the adaptive immune responses, offering new strategies of vaccinations. This review is an update of current strategies with respect to live attenuated and inactivated vaccines, DNA vaccines, viral vectors, lipid-based carrier systems such as liposomes and virosomes as well as polymeric nanoparticle vaccines and virus-like particles. In addition, this article will describe our work on a versatile and immunogenic delivery system which we have studied in the past decade and which is derived from a non-pathogenic prokaryotic organism: the “E2 scaffold” of the pyruvate dehydrogenase complex from Geobacillus stearothermophilus. PMID:26279977

  8. Pitch fractionation. Technical report

    SciTech Connect

    Weinberg, V.L.; White, J.L.

    1981-12-15

    Petroleum pitch (Ashland A240) has been subjected to thermal treatment and solvent fractionation to produce refined pitches to be evaluated as impregnants for carbon-carbon composites. The solvent fractions were obtained by sequential Soxhlet extraction with solvents such as hexane, cyclohexane, toluene, and pyridine. The most severe thermal treatment produced a mesophase pitch (approximately 50% mesophase); an appreciable portion of the mesophase was soluble in strong solvents. There were substantial differences in chemical composition and in pyrolysis behavior of the fractions. As the depth of fraction increased, the pyrolysis yield and bloating increased, and the microstructure of the coke became finer until glassy microconstituents were formed in the deepest fractions.

  9. Stool Test: H. Pylori Antigen

    MedlinePlus

    ... Your 1- to 2-Year-Old Stool Test: H. Pylori Antigen KidsHealth > For Parents > Stool Test: H. Pylori Antigen A A A What's in this article? ... en español Muestra de materia fecal: antígeno de H. pylori What It Is Helicobacter pylori ( H. pylori ) bacteria ...

  10. Dividing Fractions: A Pedagogical Technique

    ERIC Educational Resources Information Center

    Lewis, Robert

    2016-01-01

    When dividing one fraction by a second fraction, invert, that is, flip the second fraction, then multiply it by the first fraction. To multiply fractions, simply multiply across the denominators, and multiply across the numerators to get the resultant fraction. So by inverting the division of fractions it is turned into an easy multiplication of…

  11. [Molecular nature of the Brucella polysaccharide antigen (poly-B)].

    PubMed

    L'vov, V L; Pluzhnikova, G N; Lapina, E B; Shashkov, A S; Askerova, S A

    1987-08-01

    Cyclic (1----2)-beta-D-glucan was isolated from killed cells of pathogenic Brucella melitensis 16M. Its structure was deduced mainly from the acid hydrolysis, methylation analysis and 13C-NMR spectroscopy data. The cycloglucan and demicellated lipopolysaccharide of B. melitensis 16M form a stable complex identical, by immunodiffusion test, to the earlier described polysaccharide B antigen.

  12. Induction of the autoantigen proliferating cell nuclear antigen in T lymphocytes by a mycobacterial antigen.

    PubMed Central

    Haftel, H M; Chang, Y; Hinderer, R; Hanash, S M; Holoshitz, J

    1994-01-01

    Mycobacteria have been implicated in the pathogenesis of autoimmunity. To determine the potential effect of mycobacterial antigens on peripheral blood mononuclear cells (PBMC), we analyzed PBMC incubated with the acetone-precipitable fraction of Mycobacterium tuberculosis (APMT) for changes in cellular protein expression. Two-dimensional gel analysis showed induction of a 36-kD polypeptide identified as proliferating cell nuclear antigen (PCNA), a known autoantigen, after incubation with AP-MT. PCNA plays a role in cell proliferation and is expressed as a late growth regulated factor. However, its synthesis in response to AP-MT was induced as an early event. The early induction of PCNA was regulated at a posttranscriptional level and was restricted to T cells. Treatment of PBMC with known T cell mitogens, namely PHA, anti-CD3 antibodies, and staphylococcal superantigens failed to induce an early PCNA increase. The distinct characteristics of the AP-MT effect on PCNA expression suggest a separate mechanism of induction in response to AP-MT, compared with the late increase observed in response to mitogens. The induction of PCNA in response to mycobacterial antigens may represent a pathogenically relevant mechanism in autoimmunity. Images PMID:7929811

  13. Antigenic variation in Giardia lamblia.

    PubMed

    Prucca, Cesar G; Lujan, Hugo D

    2009-12-01

    Giardia lamblia undergoes antigenic variation, both in vitro and within the intestines of infected individuals. Variant-specific surface proteins (VSPs) cover the entire surface of the trophozoites and are the main antigens recognized by the host. Only 1 of about 200 VSP genes encoded by the Giardia genome is expressed on the surface of individual Giardia cells at any time; however, VSP antigen switching occurs spontaneously. In the recent year, significant advances in the knowledge of the antigen switching process have been achieved, which strongly suggests that antigenic variation in Giardia is regulated at the post-transcriptional level by a mechanism similar to RNA interference (RNAi). Several enzymes of the RNAi pathway are directly involved in VSP mRNA silencing and/or translational repression. Although several questions remain regarding how individual VSP antigens are selected for expression on the parasite surface, it is clear that an epigenetic mechanism is involved. In this review, we summarize the characteristics of this fascinating mechanism, analyse conflicting information regarding the structure of VSPs as it relates to the host's immune response, and highlight the major issues that need to be resolved to fully understand antigenic variation in this important pathogen.

  14. FRACTIONAL PEARSON DIFFUSIONS.

    PubMed

    Leonenko, Nikolai N; Meerschaert, Mark M; Sikorskii, Alla

    2013-07-15

    Pearson diffusions are governed by diffusion equations with polynomial coefficients. Fractional Pearson diffusions are governed by the corresponding time-fractional diffusion equation. They are useful for modeling sub-diffusive phenomena, caused by particle sticking and trapping. This paper provides explicit strong solutions for fractional Pearson diffusions, using spectral methods. It also presents stochastic solutions, using a non-Markovian inverse stable time change.

  15. FRACTIONAL PEARSON DIFFUSIONS

    PubMed Central

    Leonenko, Nikolai N.; Meerschaert, Mark M.

    2013-01-01

    Pearson diffusions are governed by diffusion equations with polynomial coefficients. Fractional Pearson diffusions are governed by the corresponding time-fractional diffusion equation. They are useful for modeling sub-diffusive phenomena, caused by particle sticking and trapping. This paper provides explicit strong solutions for fractional Pearson diffusions, using spectral methods. It also presents stochastic solutions, using a non-Markovian inverse stable time change. PMID:23626377

  16. Influenza A Virus Hemagglutinin Trimerization Completes Monomer Folding and Antigenicity

    PubMed Central

    Magadán, Javier G.; Khurana, Surender; Das, Suman R.; Frank, Gregory M.; Stevens, James; Golding, Hana; Bennink, Jack R.

    2013-01-01

    Influenza A virus (IAV) remains an important human pathogen largely because of antigenic drift, the rapid emergence of antibody escape mutants that precludes durable vaccination. The most potent neutralizing antibodies interact with cognate epitopes in the globular “head” domain of hemagglutinin (HA), a homotrimeric glycoprotein. The H1 HA possesses five distinct regions defined by a large number of mouse monoclonal antibodies (MAbs), i.e., Ca1, Ca2, Cb, Sa, and Sb. Ca1-Ca2 sites require HA trimerization to attain full antigenicity, consistent with their locations on opposite sides of the trimer interface. Here, we show that full antigenicity of Cb and Sa sites also requires HA trimerization, as revealed by immunofluorescence microscopy of IAV-infected cells and biochemically by pulse-chase radiolabeling experiments. Surprisingly, epitope antigenicity acquired by HA trimerization persists following acid triggering of the globular domains dissociation and even after proteolytic release of monomeric heads from acid-treated HA. Thus, the requirement for HA trimerization by trimer-specific MAbs mapping to the Ca, Cb, and Sa sites is not dependent upon the bridging of adjacent monomers in the native HA trimer. Rather, complete antigenicity of HA (and, by inference, immunogenicity) requires a final folding step that accompanies its trimerization. Once this conformational change occurs, HA trimers themselves would not necessarily be required to induce a highly diverse neutralizing response to epitopes in the globular domain. PMID:23824811

  17. A "liver" antigen associated with avian erythroblastosis: binding by bentonite and precipitation with sodium dodecyl sulphate.

    PubMed Central

    Darcel, C L

    1982-01-01

    The properties of a complement fixing antigen, EbAg, extracted from erythroblastosis-affected chicken livers are described. The antigen in extracts freed of structural protein is strongly bound by bentonite, but not by barium sulphate. Strongly alkaline solutions of sodium dodecyl sulphate are required to release the antigen from bentonite. Acidification of the detergent solution precipitates the active solution precipitates the active protein. Extraction of heme from the acidified detergent precipitate by methyl-ethyl ketone further purifies the antigen. This acid detergent treatment eliminates the need to use bentonite as a purification step. PMID:6280825

  18. Polymer blend particles with defined compositions for targeting antigen to both class I and II antigen presentation pathways.

    PubMed

    Tran, Kenny K; Zhan, Xi; Shen, Hong

    2014-05-01

    Defense against many persistent and difficult-to-treat diseases requires a combination of humoral, CD4(+) , and CD8(+) T-cell responses, which necessitates targeting antigens to both class I and II antigen presentation pathways. In this study, polymer blend particles are developed by mixing two functionally unique polymers, poly(lactide-co-glycolide) (PLGA) and a pH-responsive polymer, poly(dimethylaminoethyl methacrylate-co-propylacrylic acid-co-butyl methacrylate) (DMAEMA-co-PAA-co-BMA). Polymer blend particles are shown to enable the delivery of antigens into both class I and II antigen presentation pathways in vitro. Increasing the ratio of the pH-responsive polymer in blend particles increases the degree of class I antigen presentation, while maintaining high levels of class II antigen presentation. In a mouse model, it is demonstrate