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Sample records for acid incorporating poloxamer

  1. Acid-Labile Amphiphilic PEO-b-PPO-b-PEO Copolymers: Degradable Poloxamer Analogs.

    Worm, Matthias; Kang, Biao; Dingels, Carsten; Wurm, Frederik R; Frey, Holger

    2016-05-01

    Poly ((ethylene oxide)-b-(propylene oxide)-b-(ethylene oxide)) triblock copolymers commonly known as poloxamers or Pluronics constitute an important class of nonionic, biocompatible surfactants. Here, a method is reported to incorporate two acid-labile acetal moieties in the backbone of poloxamers to generate acid-cleavable nonionic surfactants. Poly(propylene oxide) is functionalized by means of an acetate-protected vinyl ether to introduce acetal units. Three cleavable PEO-PPO-PEO triblock copolymers (Mn,total = 6600, 8000, 9150 g·mol(-1) ; Mn,PEO = 2200, 3600, 4750 g·mol(-1) ) have been synthesized using anionic ring-opening polymerization. The amphiphilic copolymers exhibit narrow molecular weight distributions (Ð = 1.06-1.08). Surface tension measurements reveal surface-active behavior in aqueous solution comparable to established noncleavable poloxamers. Complete hydrolysis of the labile junctions after acidic treatment is verified by size exclusion chromatography. The block copolymers have been employed as surfactants in a miniemulsion polymerization to generate polystyrene (PS) nanoparticles with mean diameters of ≈200 nm and narrow size distribution, as determined by dynamic light scattering and scanning electron microscopy. Acid-triggered precipitation facilitates removal of surfactant fragments from the nanoparticles, which simplifies purification and enables nanoparticle precipitation "on demand." PMID:27000789

  2. Formulation and characterization of poloxamer 407®: thermoreversible gel containing polymeric microparticles and hyaluronic acid

    Gabriela Garrastazu Pereira

    2013-01-01

    Full Text Available The influence of the composition and preparation method on the sol-gel transition temperature (Tsol-gel and rheological response of poloxamer-based formulations was determined. Manual and more complex mechanical stirring were found to provide similar results. In addition, a linear dependence of Tsol-gel on the poloxamer content was observed in the range of concentrations analyzed, and a Poloxamer 407® concentration of 18% was selected. The addition of hyaluronic acid did not lead to significant changes in the Tsol-gel values. In contrast, the addition of microparticles caused a reduction in Tsol-gel without a significant reduction in gel strength, and pseudoplastic characteristics were observed, indicating that a thermoreversible gel was obtained with a rheology suitable for application in the treatment of burn wounds.

  3. PEGylated and poloxamer-modified chitosan nanoparticles incorporating a lysine-based surfactant for pH-triggered doxorubicin release.

    Scheeren, Laís E; Nogueira, Daniele R; Macedo, Letícia B; Vinardell, M Pilar; Mitjans, Montserrat; Infante, M Rosa; Rolim, Clarice M B

    2016-02-01

    The growing demand for efficient chemotherapy in many cancers requires novel approaches in target-delivery technologies. Nanomaterials with pH-responsive behavior appear to have potential ability to selectively release the encapsulated molecules by sensing the acidic tumor microenvironment or the low pH found in endosomes. Likewise, polyethylene glycol (PEG)- and poloxamer-modified nanocarriers have been gaining attention regarding their potential to improve the effectiveness of cancer therapy. In this context, DOX-loaded pH-responsive nanoparticles (NPs) modified with PEG or poloxamer were prepared and the effects of these modifiers were evaluated on the overall characteristics of these nanostructures. Chitosan and tripolyphosphate were selected to form NPs by the interaction of oppositely charged compounds. A pH-sensitive lysine-based amphiphile (77KS) was used as a bioactive adjuvant. The strong dependence of 77KS ionization with pH makes this compound an interesting candidate to be used for the design of pH-sensitive devices. The physicochemical characterization of all NPs has been performed, and it was shown that the presence of 77KS clearly promotes a pH-triggered DOX release. Accelerated and continuous release patterns of DOX from CS-NPs under acidic conditions were observed regardless of the presence of PEG or poloxamer. Moreover, photodegradation studies have indicated that the lyophilization of NPs improved DOX stability under UVA radiation. Finally, cytotoxicity experiments have shown the ability of DOX-loaded CS-NPs to kill HeLa tumor cells. Hence, the overall results suggest that these pH-responsive CS-NPs are highly potent delivery systems to target tumor and intracellular environments, rendering them promising DOX carrier systems for cancer therapy. PMID:26674840

  4. Characterization and micellization of a poloxamer block copolymer

    Hvidt, S.; Pedersen, Walther Batsberg

    2007-01-01

    Several poloxamers that are symmetrical EPE block copolymers (E and P are ethylene and propylene oxide, respectively) have been characterized by size exclusion chromatography on Superose columns in water. The poloxamers contain between 12 and 26 wt% of smaller-size UV-absorbing impurities....... Poloxamer P94 (E28P48E28) forms micelles with increasing temperature, and micellization was investigated by eluent gel permeation chromatography (EGPC). EGPC results demonstrate that P94 impurities are not incorporated into the micelles up to 38°C. The importance of poloxamer heterogeneity for thermodynamic...

  5. Incorporation of tritiated stearic acid in polymer

    The chemical fixation of tritium in stearic acid followed by embedding the tritiated stearic acid into an epoxy resin proved to be a good method for permanent waste disposal of tritium gas. Tc-126 and Tc-136 hardener were used for this purpose. The polymerization temperature of the polymer products was found to be highly decreased by adding stearic acid and slightly decreased by adding stearic acid or by raising the temperature of the leachant medium and slightly decreased by irradiation. On the other hand, small improvement in the compressive strength of the polymer products was achieved by adding silica powder. The polymer product showed high flame points above 573 K although it contains stearic acid. DTA and TG analysis indicate that the thermal stability of polymer products containing stearic acid and silica powder is better than samples containing only stearic acid. Stearic acid was found to possess good resistance to irradiation and the amount of hydrogen and oxygen gases measured due to irradiation of the epoxy resin products containing stearic acid was found to be negligible. The leaching rate of tritium from the epoxy resin sample containing tritiated stearic acid was measured under different experimental conditions. The dominant factors affecting the leaching rate of tritium are the leachant medium and its temperature. From the above results it can be generally recommended to incorporate tritiated stearic acid in the epoxy resin Tc-126 alone without silica powder and to avoid great rise in the temperature of the leachant medium. Under optimum conditions excellent results can be achieved for permanent waste disposal of tritium gas and about 1.5.1016 Bq tritium gas could be fixed on about 1 kg Tc-126 epoxy resin. (author) 16 figs., 3 tabs., 11 refs

  6. Electrodelivery of Drugs into Cancer Cells in the Presence of Poloxamer 188

    Iana Tsoneva

    2010-01-01

    Full Text Available In the present study it is shown that poloxamer 188, added before or immediately after an electrical pulse used for electroporation, decreases the number of dead cells and at the same time does not reduce the number of reversible electropores through which small molecules (cisplatin, bleomycin, or propidium iodide can pass/diffuse. It was suggested that hydrophobic sections of poloxamer 188 molecules are incorporated into the edges of pores and that their hydrophilic parts act as brushy pore structures. The formation of brushy pores may reduce the expansion of pores and delay the irreversible electropermeability. Tumors were implanted subcutaneously in both flanks of nude mice using HeLa cells, transfected with genes for red fluorescent protein and luciferase. The volume of tumors stopped to grow after electrochemotherapy and the use of poloxamer 188 reduced the edema near the electrode and around the subcutaneously growing tumors.

  7. Mutagenic effect of incorporated tritium amino acids

    Genetic effect of tritium labelled amino acids was studied. The experiments were carried out on white mongreal rats, genetic effects were evaluated by dominant lethal mutation frequency in male germ cells. It was shown that administration of tritium amino acids results in genetic violations in male germ cells manifested in progeny death. Assessment of integral temporal indices of induced post implantation embryos death showed that 3H-lysine effect exceeds tritium oxide effect by 1.5-2 fold in case of equal absorbed doses. The obtained results are used in alculation of radiation hygienic standards for biogenic tritium compounds. 4 refs.; 1 tab

  8. Method Development for Efficient Incorporation of Unnatural Amino Acids

    Harris, Paul D.

    2014-04-01

    The synthesis of proteins bearing unnatural amino acids has the potential to enhance and elucidate many processes in biochemistry and molecular biology. There are two primary methods for site specific unnatural amino acid incorporation, both of which use the cell’s native protein translating machinery: in vitro chemical acylation of suppressor tRNAs and the use of orthogonal amino acyl tRNA synthetases. Total chemical synthesis is theoretically possible, but current methods severely limit the maximum size of the product protein. In vivo orthogonal synthetase methods suffer from the high cost of the unnatural amino acid. In this thesis I sought to address this limitation by increasing cell density, first in shake flasks and then in a bioreactor in order to increase the yield of protein per amount of unnatural amino acid used. In a parallel project, I used the in vitro chemical acylation system to incorporate several unnatural amino acids, key among them the fluorophore BODIPYFL, with the aim of producing site specifically fluorescently labeled protein for single molecule FRET studies. I demonstrated successful incorporation of these amino acids into the trial protein GFP, although incorporation was not demonstrated in the final target, FEN1. This also served to confirm the effectiveness of a new procedure developed for chemical acylation.

  9. Incorporation of conjugated linoleic acid (CLA and α-linolenic acid (LNA in pacu fillets

    Deoclécio José Barilli

    2014-03-01

    Full Text Available The objective of this study was to evaluate the incorporation of conjugated linoleic acid and α-linolenic acid in fillets of pacu fish raised in net cages and fed diets enriched with these acids. The fish were fed for 49 days, and at the end of this period the fatty acid content in the fillets was determined by gas chromatography. Concentrations of α-linolenic acid, eicosapentaenoic acid, and the total omega-3 (n-3 fatty acid in the fillets increased, improving the n-6/n-3 ratio. In addition, the incorporation of conjugated linoleic acid in the fish fillets proved well established. This study showed that the use of diets enriched with conjugated linoleic acid and α-linolenic acid results in the incorporation of these acids in the of pacu fish fillets, improving their nutritional quality.

  10. Tuning hardness in calcite by incorporation of amino acids

    Kim, Yi-Yeoun; Carloni, Joseph D.; Demarchi, Beatrice; Sparks, David; Reid, David G.; Kunitake, Miki E.; Tang, Chiu C.; Duer, Melinda J.; Freeman, Colin L.; Pokroy, Boaz; Penkman, Kirsty; Harding, John H.; Estroff, Lara A.; Baker, Shefford P.; Meldrum, Fiona C.

    2016-08-01

    Structural biominerals are inorganic/organic composites that exhibit remarkable mechanical properties. However, the structure-property relationships of even the simplest building unit--mineral single crystals containing embedded macromolecules--remain poorly understood. Here, by means of a model biomineral made from calcite single crystals containing glycine (0-7 mol%) or aspartic acid (0-4 mol%), we elucidate the origin of the superior hardness of biogenic calcite. We analysed lattice distortions in these model crystals by using X-ray diffraction and molecular dynamics simulations, and by means of solid-state nuclear magnetic resonance show that the amino acids are incorporated as individual molecules. We also demonstrate that nanoindentation hardness increased with amino acid content, reaching values equivalent to their biogenic counterparts. A dislocation pinning model reveals that the enhanced hardness is determined by the force required to cut covalent bonds in the molecules.

  11. Incorporation of oxygen into abscisic acid and phaseic acid for molecular oxygen

    Abscisic acid accumulates in detached, wilted leaves of Xanthium strumariu. When these leaves are subsequently rehydrated, phaseic acid, a catabolite of abscisic acid, accumulates. Analysis by gas chromatography-mass spectrometry of phaseic acid isolated from stressed and subsequently rehydrated leaves placed in an atmosphere containing 20% 18O2 and 80% N2 indicates that one atom of 18O is incorporated in the 6'-hydroxymethyl group of phaseic acid. This suggests that the enzyme that converts abscisic acid to phaseic acid is an oxygenase. Analysis by gas chromatography-mass spectrometry of abscisic acid isolated from stressed leaves kept in an atmosphere containing 18O2 indicates that one atom of 18O is presented in the carboxyl group of abscisic acid. Thus, when abscisic acid accumulates in water-streesed leaves, only one of the four oxygens present in the abscisic acid molecule is derived from molecular oxygen. This suggest that either (a) the oxygen present in the 1'-, 4'-, and one of the two oxygens at the 1-position of abscisic acid arise from water, or (b) there exists a stored precursor with oxygen atoms already present in the 1'- and 4'-positions of abscisic acid which is converted to abscisic acid under conditions of water stress. 17 references, 2 figures, 1 tables

  12. [Effect of proteolysis inhibitors on the incorporation of labelled amino acids into proteins].

    Konikova, A S; Korotkina, R N

    1975-01-01

    Role of peptide bond breaks in the incorporation of amino acids into proteins in a "protein--amino acid" system is investigated. For this purpose the incorporation of labelled amino acids into trypsin under the inhibition of its autolysis by a specific inhibitor from soybean and epsilon-amino-caproic acid is studied. The trypsin inhibitor from soybean is found to suppress considerably the incorporation of 14C-glycine, 14C-lysine and 14C-methionine into crystal trypsin and not to affect the incorporation of labelled amino acids into chomotrypsin, papain and carboxypeptidase. Epsilon-Aminocaproic acid inhibited 14C-glycine incorporation into crystal trypsin by 40% and did not change its incorporation level into serum albumin. The dependency of amino acid incorporation level into trypsin on the activity of autolysis in the "protein--amino acid" system is demonstrated. PMID:1212456

  13. Lipid corralling and poloxamer squeeze-out in membranes

    Wu, G.H.; Majewski, J.; Ege, C.;

    2004-01-01

    Using x-ray scattering measurements we have quantitatively determined the effect of poloxamer 188 (P188), a polymer known to seal damaged membranes, on the structure of lipid monolayers. P188 selectively inserts into low lipid-density regions of the membrane and "corrals" lipid molecules to pack ...

  14. Effects of Poloxamer 188 on human PMN cells

    Harting, Matthew T.; Jimenez, Fernando; Kozar, Rosemary A.; Moore, Frederick A.; Mercer, David W.; Hunter, Robert L.; Cox, Charles S.; Gonzalez, Ernest A.

    2008-01-01

    Poloxamer 188 (P188), a nonionic block copolymer chemical surfactant known to have cytoprotective, rheologic, anti-inflammatory, and anti-thrombotic activity, has shown promise in the management of selected trauma patients. We studied human PMN oxidative burst and adhesion molecule expression when exposed to P188.

  15. Selective incorporation of docosahexaenoic acid into lysobisphosphatidic acid in cultured THP-1 macrophages.

    Besson, Nelly; Hullin-Matsuda, Francoise; Makino, Asami; Murate, Motohide; Lagarde, Michel; Pageaux, Jean-Francois; Kobayashi, Toshihide; Delton-Vandenbroucke, Isabelle

    2006-02-01

    Lysobisphosphatidic acid (LBPA) is highly accumulated in specific domains of the late endosome and is involved in the biogenesis and function of this organelle. Little is known about the biosynthesis and metabolism of this lipid. We examined its FA composition and the incorporation of exogenous FA into LBPA in the human monocytic leukemia cell line THP-1. The LBPA FA composition in THP-1 cells exhibits an elevated amount of oleic acid (18:1n-9) and enrichment of PUFA, especially DHA (22:6n-3). DHA supplemented to the medium was efficiently incorporated into LBPA. In contrast, arachidonic acid (20:4n-6) was hardly esterified to LBPA under the same experimental conditions. The turnover of DHA in LBPA was similar to that in other phospholipids. Specific incorporation of DHA into LBPA was also observed in baby hamster kidney fibroblasts, although LBPA in these cells contains very low endogenous levels of DHA in normal growth conditions. Our resuIts, together with published observations, suggest that the specific incorporation of DHA into LBPA is a common phenomenon in mammalian cells. The physiological significance of DHA-enriched LBPA is discussed. PMID:17707985

  16. Long-term incorporation of tritiated adenine into deoxyribonucleic acid and ribonucleic acid by Treponema pallidum (Nichols strain).

    Norris, S J; Miller, J N; Sykes, J A

    1980-01-01

    Treponema pallidum (Nichols strain), extracted in medium containing Eagle minimal essential medium 50% fresh, heat-inactivated normal rabbit serum, and 1.0 mM dithiothreitol, was incubated under 3% oxygen in the presence of tritiated nucleic acid precursors. [8-3H]adenine was incorporated with high efficiency into trichloroacetic acid-insoluble material; 2'-deoxyadenosine and uridine were incorporated in lower quantities, and thymine and thymidine were not incorporated. Incorporation of [3H]a...

  17. Incorporation of Palmitic Acid or Stearic Acid into Soybean Oils Using Enzymatic Interesterification.

    Teh, Soek Sin; Voon, Phooi Tee; Hock Ong, Augustine Soon; Choo, Yuen May

    2016-09-01

    Incorporations of nature fatty acids which were palmitic acid and stearic acid into the end positions of soybean oils were done using sn-1,3 specific immobilised lipase from Rhizomucor miehei at different ratios in order to produce symmetrical triglycerides without changing the fatty acids at sn-2 position. The optimum ratio for the process was 25:75 w/w. There were 19.2% increase of SFA for P25 and 16% increase for S25 at the sn-1,3 positions. The research findings indicated that the structured lipids produced from enzymatic interesterification possessed a higher oxidative stability than soybean oil. The newly formed structured lipids (SUS type) could be good sources for various applications in food industry. PMID:27477075

  18. In vivo incorporation of labeled fatty acids in rat liver lipids after oral administration

    Striking differences were found in the compartmentalization of fatty acids into liver lipid fractions. The saturated fatty acids--lauric, myristic, palmitic and stearic--were incorporated into phosphoglycerides at faster rates with increasing chain lengths, while triglyceride incorporation was almost uniform. The degree of incorporation of the unsaturated fatty acids into phosphoglycerides (structural) compared to triglyceride (storage and energy) was the converse of their oxidation rates. The incorporation of oleic, linoleic and alpha-linolenic acids was mainly into triglyceride, whereas dihomo-gamma-linolenic acid and arachidonic acid were preferentially incorporated into phosphoglycerides. The data suggest that distribution of each fatty acid is different depending on its destination for structural or energy function

  19. In vivo incorporation of labeled fatty acids in rat liver lipids after oral administration

    Leyton, J.; Drury, P.J.; Crawford, M.A.

    1987-08-01

    Striking differences were found in the compartmentalization of fatty acids into liver lipid fractions. The saturated fatty acids--lauric, myristic, palmitic and stearic--were incorporated into phosphoglycerides at faster rates with increasing chain lengths, while triglyceride incorporation was almost uniform. The degree of incorporation of the unsaturated fatty acids into phosphoglycerides (structural) compared to triglyceride (storage and energy) was the converse of their oxidation rates. The incorporation of oleic, linoleic and alpha-linolenic acids was mainly into triglyceride, whereas dihomo-gamma-linolenic acid and arachidonic acid were preferentially incorporated into phosphoglycerides. The data suggest that distribution of each fatty acid is different depending on its destination for structural or energy function.

  20. Incorporated fish oil fatty acids prevent action potential shortening induced by circulating fish oil fatty acids

    Hester M Den Ruijter

    2010-11-01

    Full Text Available Increased consumption of fatty fish, rich in omega-3 polyunsaturated fatty acids (3-PUFAs reduces the severity and number of arrhythmias. Long term 3-PUFA-intake modulates the activity of several cardiac ion channels leading to cardiac action potential shortening. Circulating 3-PUFAs in the bloodstream and incorporated 3-PUFAs in the cardiac membrane have a different mechanism to shorten the action potential. It is, however, unknown whether circulating 3-PUFAs in the bloodstream enhance or diminish the effects of incorporated 3-PUFAs. In the present study, we address this issue. Rabbits were fed a diet rich in fish oil (3 or sunflower oil (9, as control for 3 weeks. Ventricular myocytes were isolated by enzymatic dissociation and action potentials were measured using the perforated patch clamp technique in the absence and presence of acutely administered 3-PUFAs. Plasma of 3 fed rabbits contained more free eicosapentaenoic acid (EPA and isolated myocytes of 3 fed rabbits contained higher amounts of both EPA and docosahexaenoic acid (DHA in their sarcolemma compared to control. In the absence of acutely administered fatty acids, 3 myocytes had a shorter action potential with a more negative plateau than 9 myocytes. In the 9 myocytes, but not in the 3 myocytes, acute administration of a mixture of EPA+DHA shortened the action potential significantly. From these data we conclude that incorporated 3-PUFAs into the sarcolemma and acutely administered 3 fatty acids do not have a cumulative effect on action potential duration and morphology. As a consequence, patients with a high cardiac 3-PUFA status will probably not benefit from short term 3 supplementation as an antiarrhythmic therapy.

  1. Incorporation and excision of 5-fluorouracil from deoxyribonucleic acid in Escherichia coli.

    Warner, H. R.; Rockstroh, P A

    1980-01-01

    When Escherichia coli are grown in the presence of 5-fluorouracil, the 5-fluorouracil is incorporated almost exclusively into ribonucleic acid as fluorouridylate. In this study, small but detectable amounts were incorporated into ribonucleic acid as fluorocytidylate and into deoxyribonucleic acid as fluorodeoxyuridylate and fluorodeoxycytidylate. The amount of 5-fluorouracil found in deoxyribonucleic acid as fluorodeoxyuridylate increased 50-fold when the cells were deficient in both deoxyuri...

  2. Incorporation of conjugated linoleic acid and vaccenic acid into lipids from rat tissues and plasma

    Lund, Pia; Sejrsen, Kristen; Straarup, Ellen Marie

    2006-01-01

    The objective of this study was to determine the incorporation of conjugated linoleic acid (CLA) into triacylglycerols (TAG) and phospholipids (PL) of tissues and plasma, and to interpret the role of dietary-derived vaccenic acid (VA) in increasing the tissue content of CLA (c9,t11) and the...... influence on the fatty acid profile. We fed five groups of rats semi-purified diets with varying levels of CLA and VA: control butter with low CLA (c9,t11) and VA; control butter added 5% CLA (c9,t11); control butter added 5% Tonalin [equal amount of CLA (c9,t11) and CLA (t10,c12)]; control butter added 5...

  3. Enzymatically mediated incorporation of 2-chlorophenol 4-chlorophenol into humic acids

    Lassen, P.; Randall, A.; Jørgensen, O.;

    1994-01-01

    A possible route to chlorinated humic substances in the environment, is an indirect chlorination of humic material by enzymatically mediated incorporation of low molecular weight organo-chlorine compounds into the humic skeleton. The enzymatically mediated incorporation of 2-chlorophenol and 4......-chlorophenol into humic acids by Horseradish Peroxidase is reported. The incorporation is accompanied by a significant polymerization of the chlorophenols. The stability of the chlorinated humic acids as well as the environmental implication are discussed....

  4. Melittin liposomes surface modified with poloxamer 188: in vitro characterization and in vivo evaluation.

    Tian, J L; Ke, X; Chen, Z; Wang, C J; Zhang, Y; Zhong, T C

    2011-05-01

    Melittin liposomes surface modified with poloxamer 188 were developed, and the effect of poloxamer 188 was investigated with regard to anti-cancer effect and vascular stimulation. Melittin liposomes surface modified with poloxamer 188 at different concentrations (0%, 2%, and 5%) were prepared using the adsorption method, followed by in vitro characterization, including entrapment efficiency, zeta potential, particle size, and morphology. Subsequently, the influence of repeated freeze-thawing on the liposomes was investigated, and the effect of poloxamer 188 on the repeated freeze-thawing process was explored. Vascular stimulation effects of MLT, and MLT liposome that surface coated with or without poloxamer were all studied. Pharmacokinetics of the different MLT preparations were determined and the anticancer activity of the MLT formulations was investigated. The particle size of the liposomes gradually increased with increasing poloxamer 188 content, while the entrapment efficiency did not change significantly. After the first freeze-thaw cycle, size and PDI were both markedly reduced, entrapment efficiency rose, and there was no significant change of zeta potential. The vascular irritation caused by MLT could be reduced to an extent by encapsulation in liposome, but not completely eliminated, while liposomes coated with poloxamer 188 can effectively abolish the phenomenon. Melittin liposomes with surface modified by poloxamer exhibit enhanced bioavailability, effective anticancer activity, and reduced side effects compared with melittin solution. Poloxamer plays an important role in melittin liposomes. PMID:21699070

  5. Benzothiazole incorporated barbituric acid derivatives: synthesis and anticonvulsant screening.

    Siddiqui, Nadeem; Ahsan, Waquar

    2009-08-01

    A series of 1-(6-substituted-1,3-benzothiazol-2-yl)-3-(substituted phenyl)hexahydro-2,4,6-pyrimidinetriones 4a-t were synthesized starting from substituted anilines. These compounds contained two active anticonvulsant pharmacophores, benzothiazole and barbituric acid. Structures of the compounds were confirmed on the basis of different spectroscopic techniques. All the compounds were evaluated for their anticonvulsant activity. Three compounds 4c, 4d, and 4s showed promising anticonvulsant activities in Maximal Electroshock Seizure test (MES) and subcutaneous pentylenetetrazole test (scPTZ). They also displayed a wide safety profile when tested for the minimal motor impairment test. PMID:19565603

  6. Incorporation of 15N-inorganic nitrogen into free-amino acids in germinating corn

    Incorporation of 15N-labeled compounds, (K15NO3) and (15NH4)2SO4, into free-amino acids was measured in germinating corn. Sterilized seeds of sweet corn (Choko No. 865) were sown on the filter papers soaked in 10 ml of the solution containing one of the labeled compounds (40 ppm N, 99 atom % excess) in petri dishes and germinated at 30 deg C. After 48 hours and 72 hours, 15N-incorporation was measured in 5 seedlings selected owing to uniform growth. A GC-MS was used for measuring the ratio of 15N isotopes present in free-amino acids. 15N incorporation into free-amino acids hardly occurred when corn was germinated in the solution containing K15NO3, which suggested that endogenous nitrogen was used during the early germination stage of corn when nitrate is present. Incorporation into amino acids was greater when corn was germinated in the medium containing (15NH4)2SO4, than the case of the solution containing K15NO3. When corn was germinated in the solution containing (15NH4)2SO4, assimilation of 15N into asparagine or aspartic acid was comparatively higher than that into the other amino acids, though the incorporation rate was low. Thus, in intact germinating corn, the hydrolyzed product of protein was utilized for germination with priority, and dependence on exogenous nitrogen was low. (Kaihara, S.)

  7. Incorporation and profile of fatty acids in tilapia fillets (Oreochromis niloticus fed with tung oil

    Elton Guntendorfer Bonafé

    2013-02-01

    Full Text Available The acceptance of tung oil enriched diet and the incorporation of conjugated linolenic acid - CLnA into fillets of Genetically Improved Farmed Tilapia (GIFT were investigated. The diet was well accepted, and after 10 days CLnA was incorporated into the fillets with a 1.02% content of total fatty acids (FA. In addition, biosynthesis of the conjugated linoleic acid isomers - CLA (0.31% of fillet total FA content from CLnA, and the presence of alpha-linolenic acid - LNA (1.08% of fillet total FA content, eicosapentaenoic acid - EPA (2.85% of fillet total FA content and docosahexaenoic acid - DHA (3.08% of fillet total FA content were observed. Therefore, the consumption of this fish can increase the intake of different FA (CLnA, CLA, LNA, EPA and DHA, which play an important role in human metabolism.

  8. Nanoparticle agglomerates of indomethacin: The role of poloxamers and matrix former on their dissolution and aerosolisation efficiency.

    Malamatari, Maria; Somavarapu, Satyanarayana; Bloxham, Mark; Buckton, Graham

    2015-11-10

    Nanoparticles (NPs) were prepared and assembled to microsized agglomerates with and without matrix formers (mannitol and L-leucine) by coupling wet milling and spray drying to harmonise the advantages of NPs with handling and aerodynamics of microparticles without induction of amorphisation. Indomethacin was selected as poorly water-soluble drug and poloxamers with different ratios of hydrophilic to hydrophobic domains were evaluated as stabilisers comparatively to D-α-Tocopherol polyethylene-glycol succinate (TPGS). Particle size of nanosuspensions and morphology, size, crystal form, drug loading, redispersibility, in vitro dissolution, and in vitro aerosolisation of NP-agglomerates were determined. Molecular weight of stabilisers affected the rate but not the limit of NP size reduction and the length of hydrophilic segment in poloxamers was found important for the nanosuspension stabilisation. SEM revealed the structure of agglomerates consisting of nanocrystal assemblies. XRPD with DSC proved that NP agglomerates retained their crystallinity. NP-agglomerates exhibited enhanced dissolution compared to physical mixtures of drug and stabilisers while incorporation of matrix formers enabled redispersibility upon hydration and further increased the drug dissolution. Also, matrix formers resulted in significantly improved aerosolisation with higher fine particle fractions (49-62%) and smaller mass median aerodynamic diameters (<3.5 μm), compared to cases without matrix formers (34-43% and <4.5 μm). PMID:26364709

  9. Cisplatin-incorporated nanoparticles of poly(acrylic acid-co-methyl methacrylate copolymer

    Lee KD

    2013-08-01

    Full Text Available Kyung Dong Lee,1,* Young-Il Jeong,2,* Da Hye Kim,3,4 Gyun-Taek Lim,2 Ki-Choon Choi5 1Department of Oriental Medicine Materials, Dongshin University, Naju, South Korea; 2Department of Polymer Engineering, Chonnam National University, Gwangju, South Korea; 3Faculty of Life and Environmental Science, Shimane University, Matsue, Japan; 4United Graduate School of Agricultural Sciences, Tottori University, Tottori, Japan; 5Grassland and Forages Division, National Institute of Animal Science, Rural Development Administration, Cheonan, South Korea *These authors contributed equally to this work Background: Although cisplatin is extensively used in the clinical field, its intrinsic toxicity limits its clinical use. We investigated nanoparticle formations of poly(acrylic acid-co-methyl methacrylate (PAA-MMA incorporating cisplatin and their antitumor activity in vitro and in vivo. Methods: Cisplatin-incorporated nanoparticles were prepared through the ion-complex formation between acrylic acid and cisplatin. The anticancer activity of cisplatin-incorporated nanoparticles was assessed with CT26 colorectal carcinoma cells. Results: Cisplatin-incorporated nanoparticles have small particle sizes of less than 200 nm with spherical shapes. Drug content was increased according to the increase of the feeding amount of cisplatin and acrylic acid content in the copolymer. The higher acrylic acid content in the copolymer induced increase of particle size and decrease of zeta potential. Cisplatin-incorporated nanoparticles showed a similar growth-inhibitory effect against CT26 tumor cells in vitro. However, cisplatin-incorporated nanoparticles showed improved antitumor activity against an animal tumor xenograft model. Conclusion: We suggest that PAA-MMA nanoparticles incorporating cisplatin are promising carriers for an antitumor drug-delivery system. Keywords: cisplatin, nanoparticle, poly(acrylic acid-co-methyl methacrylate, ion complexes

  10. Site specific incorporation of heavy atom-containing unnatural amino acids into proteins for structure determination

    Xie, Jianming; Wang, Lei; Wu, Ning; Schultz, Peter G.

    2008-07-15

    Translation systems and other compositions including orthogonal aminoacyl tRNA-synthetases that preferentially charge an orthogonal tRNA with an iodinated or brominated amino acid are provided. Nucleic acids encoding such synthetases are also described, as are methods and kits for producing proteins including heavy atom-containing amino acids, e.g., brominated or iodinated amino acids. Methods of determining the structure of a protein, e.g., a protein into which a heavy atom has been site-specifically incorporated through use of an orthogonal tRNA/aminoacyl tRNA-synthetase pair, are also described.

  11. Incorporation of 14C-linoleic acid in cerebrosides of psoriatic and normal human skin

    It was shown that the incorporation of 14C-linoleic acid in cerebrosides of normal and psoriatic human skin is different. In psoriatic epidermis and corium the turnover of this fatty acid is significantly elevated. It is suggested that in psoriasis the epidermal cell is not able to build up a regular carbohydrate sequences of lipids because the false carbohydrate chain activates the degradation of glycolipids and in compensating for the increased degradation raises the synthesis rate of glycolipids. (orig./MG)

  12. Orthogonal translation components for the in vivo incorporation of unnatural amino acids

    Schultz, Peter G.; Alfonta, Lital; Chittuluru, Johnathan R.; Deiters, Alexander; Groff, Dan; Summerer, Daniel; Tsao, Meng -Lin; Wang, Jiangyun; Wu, Ning; Xie, Jianming; Zeng, Huaqiang; Seyedsayamdost, Mohammad; Turner, James

    2015-08-11

    The invention relates to orthogonal pairs of tRNAs and aminoacyl-tRNA synthetase that can incorporate unnatural amino acid into proteins produced in eubacterial host cells such as E. coli, or in a eukaryotic host such as a yeast cell. The invention provides, for example but not limited to, novel orthogonal synthetases, methods for identifying and making the novel synthetases, methods for producing proteins containing unnatural amino acids, and translation systems.

  13. Orthogonal translation components for the in vivo incorporation of unnatural amino acids

    Schultz, Peter G.; Xie, Jianming; Zeng, Huaqiang

    2012-07-10

    The invention relates to orthogonal pairs of tRNAs and aminoacyl-tRNA synthetases that can incorporate unnatural amino acids into proteins produced in eubacterial host cells such as E. coli, or in a eukaryotic host such as a yeast cell. The invention provides, for example but not limited to, novel orthogonal synthetases, methods for identifying and making the novel synthetases, methods for producing proteins containing unnatural amino acids, and translation systems.

  14. Incorporation of medium chain fatty acids into fish oil triglycerides by chemical and enzymatic inter esterification

    Feltes, M. M. C.; Oliveira de Pilot, L.; Gomes Correira, F.; Grimaldi, R.; Mara Block, J.; Ninow, J. L.

    2009-07-01

    Structured triglycerides (STs) containing both medium chain fatty acids (MCFA) and polyunsaturated fatty acids (PUFA) in the same molecule offer nutritional and therapeutic benefits. The aim of this work was to establish the incorporation of MCFA into fish oil triglycerides (TAGs), while maintaining substantial levels of docosahexaenoic and eicosapentaenoic acids. The effects of different acyl donors (capric acid methyl ester/MeC10 or medium chain triglyceride/TCM) and of the catalyst (chemical or enzymatic) on the fatty acid composition of the reaction product were studied. The fatty acid composition of the fish oil TAG was modified after inter esterification to contain MCFA, and it depended on the catalyst and on the substrates. Thermo grams obtained by Differential Scanning Calorimetry (DSC) showed that inter esterification promoted noteworthy changes in the melting profile of the samples. STs of clinical nutrition interest containing both EPA and DHA obtained from fish oil along with MCFA were successfully produced. (Author) 70 refs.

  15. Main Chain Noncentrosymmetric Hydrogen Bonded Macromolecules Incorporating Aniline, Alkanol, and Alkanoic Acid Hydrogen Bond Donors

    Jeremy R. Wolf

    2014-01-01

    The syntheses and characterization of three noncentrosymmetric main chain hydrogen bonded macromolecules which incorporate aniline, alkanoic acid, and alkanol hydrogen bond donor units are reported. These macromolecules participate in weak intermolecular hydrogen bonding as demonstrated using attenuated total reflectance (ATR) FTIR. The phase transitions of these macromolecules depend on the identity of the hydrogen bond donor.

  16. An AMBER/DYANA/MOLMOL Phosphorylated Amino Acid Library Set and Incorporation into NMR Structure Calculations

    Protein structure determination using Nuclear Magnetic Resonance (NMR) requires the use of molecular dynamics programs that incorporate both NMR experimental and implicit atomic data. Atomic parameters for each amino acid type are encoded in libraries used by structure calculation programs such as DYANA and AMBER. However, only a few non-standard amino acid library sets are included in these programs or the molecular visualization program MOLMOL. Our laboratory is calculating the phosphorylated and non-phosphorylated states of peptides and proteins using NMR methods. To calculate chemically correct structures, we have extended the available molecular libraries for these programs to include the modified amino acids phosphoserine, phosphothreonine, and phosphotyrosine

  17. Amino acid isotope incorporation and enrichment factors in Pacific bluefin tuna, Thunnus orientalis.

    Bradley, Christina J; Madigan, Daniel J; Block, Barbara A; Popp, Brian N

    2014-01-01

    Compound specific isotopic analysis (CSIA) of amino acids has received increasing attention in ecological studies in recent years due to its ability to evaluate trophic positions and elucidate baseline nutrient sources. However, the incorporation rates of individual amino acids into protein and specific trophic discrimination factors (TDFs) are largely unknown, limiting the application of CSIA to trophic studies. We determined nitrogen turnover rates of individual amino acids from a long-term (up to 1054 days) laboratory experiment using captive Pacific bluefin tuna, Thunnus orientalis (PBFT), a large endothermic pelagic fish fed a controlled diet. Small PBFT (white muscle δ(15)N∼11.5‰) were collected in San Diego, CA and transported to the Tuna Research and Conservation Center (TRCC) where they were fed a controlled diet with high δ(15)N values relative to PBFT white muscle (diet δ(15)N∼13.9‰). Half-lives of trophic and source amino acids ranged from 28.6 to 305.4 days and 67.5 to 136.2 days, respectively. The TDF for the weighted mean values of amino acids was 3.0 ‰, ranging from 2.2 to 15.8 ‰ for individual combinations of 6 trophic and 5 source amino acids. Changes in the δ(15)N values of amino acids across trophic levels are the underlying drivers of the trophic (15)N enrichment. Nearly all amino acid δ(15)N values in this experiment changed exponentially and could be described by a single compartment model. Significant differences in the rate of (15)N incorporation were found for source and trophic amino acids both within and between these groups. Varying half-lives of individual amino acids can be applied to migratory organisms as isotopic clocks, determining the length of time an individual has spent in a new environment. These results greatly enhance the ability to interpret compound specific isotope analyses in trophic studies. PMID:24465724

  18. Delivery of siRNA Using Cationic Liposomes Incorporating Stearic Acid-modified Octa-Arginine.

    Yang, Dongsheng; Li, Yuhuan; Qi, Yuhang; Chen, Yongzhen; Yang, Xuewei; Li, Yujing; Liu, Songcai; Lee, Robert J

    2016-07-01

    Cationic liposomes incorporating stearic acid-modified octa-arginine (StA-R8) were evaluated for survivin small interfering RNA (siRNA) delivery. StA-R8 was synthesized and incorporated into liposomes. The composition of liposomes was optimized. Physicochemical properties, cytotoxicity, cellular uptake and gene silencing activity of the liposomes complexed to survivin siRNA were investigated. The results showed that StA-R8-containing liposomes had reduced cytotoxicity and improved delivery efficiency of siRNA into cancer cells compared with StA-R8 by itself. PMID:27354583

  19. Rice straw incorporated just before soil flooding increases acetic acid formation and decreases available nitrogen

    Ronaldir Knoblauch

    2014-02-01

    Full Text Available Incorporation of rice straw into the soil just before flooding for water-seeded rice can immobilize mineral nitrogen (N and lead to the production of acetic acid harmful to the rice seedlings, which negatively affects grain yield. This study aimed to evaluate the formation of organic acids and variation in pH and to quantify the mineral N concentration in the soil as a function of different times of incorporation of rice straw or of ashes from burning the straw before flooding. The experiment was carried out in a greenhouse using an Inceptisol (Typic Haplaquept soil. The treatments were as follows: control (no straw or ash; incorporation of ashes from previous straw burning; rice straw incorporated to drained soil 60 days before flooding; straw incorporated 30 days before flooding; straw incorporated 15 days before flooding and straw incorporated on the day of flooding. Experimental units were plastic buckets with 6.0 kg of soil. The buckets remained flooded throughout the trial period without rice plants. Soil samples were collected every seven days, beginning one day before flooding until the 13th week of flooding for determination of mineral N- ammonium (NH4+ and nitrate (NO3-. Soil solution pH and concentration of organic acids (acetic, propionic and butyric were determined. All NO3- there was before flooding was lost in approximately two weeks of flooding, in all treatments. There was sigmoidal behavior for NH4+ formation in all treatments, i.e., ammonium ion concentration began to rise shortly after soil flooding, slightly decreased and then went up again. On the 91st day of flooding, the NH4+ concentrations in soil was 56 mg kg-1 in the control treatment, 72 mg kg-1 for the 60-day treatment, 73 mg kg-1 for the 30-day treatment and 53 mg kg-1 for the ash incorporation treatment. These ammonium concentrations correspond to 84, 108, 110 and 80 kg ha-1 of N-NH4+, respectively. When the straw was incorporated on the day of flooding or 15 days

  20. Incorporation of conjugated linoleic and alpha linolenic fatty acids into Pimedolus maculatus fillets

    Ana Beatriz Zanqui

    2013-09-01

    Full Text Available The objective of this study was to investigate the fatty acid composition of Pimedolus maculatus fillets of fish fed with different diets. The fish were submitted to an adaptation period (over 30 days and fed only a control diet (diet I. After this period, the fish were divided into two groups: one group received a diet enriched with flaxseed oil (diet II, and the other received a diet enriched with conjugated linoleic acid (CLA (diet III. Subsequent collections were undertaken after 25 and 50 days. Compared to the control samples, fish fed diet II showed an increase in concentrations of total fatty acid omega-3 (n-3 in the fillets, with an improvement ratio of n-6/n-3. The incorporation of CLA into fish fed diet III was successful. Supplementing the diet of Pimedolus maculatus with CLA and flaxseed oil led to improvements in nutritional quality and fatty acid profile of the fish fillets.

  1. Incorporation of 14C-amino acids into rat hypothalamus in vitro

    Biosynthesis of peptides in vitro in male rat hypothalamic fragments was investigated by incubating the tissue with 14C-labelled leucine, proline and glycine. Depending on the amino acid used, 4 or 6 labelled peptides were extracted from the tissue fragments with organic solvent mixture. The incorporation of the labelled amino acids into peptides was time-dependent. LH-RH and oxytocin thus synthesized were identified by thin layer chromatography and high voltage electrophoresis using cold LH-RH and oxytocin as standards. (author)

  2. β-Puromycin selection of modified ribosomes for in vitro incorporation of β-amino acids.

    Dedkova, Larisa M; Fahmi, Nour Eddine; Paul, Rakesh; del Rosario, Melissa; Zhang, Liqiang; Chen, Shengxi; Feder, Glen; Hecht, Sidney M

    2012-01-10

    Ribosomally mediated protein biosynthesis is limited to α-L-amino acids. A strong bias against β-L-amino acids precludes their incorporation into proteins in vivo and also in vitro in the presence of misacylated β-aminoacyl-tRNAs. Nonetheless, earlier studies provide some evidence that analogues of aminoacyl-tRNAs bearing β-amino acids can be accommodated in the ribosomal A-site. Both functional and X-ray crystallographic data make it clear that the exclusion of β-L-amino acids as participants in protein synthesis is a consequence of the architecture of the ribosomal peptidyltransferase center (PTC). To enable the reorganization of ribosomal PTC architecture through mutagenesis of 23S rRNA, a library of modified ribosomes having modifications in two regions of the 23S rRNA (2057-2063 and 2496-2507 or 2582-2588) was prepared. A dual selection procedure was used to obtain a set of modified ribosomes able to carry out protein synthesis in the presence β-L-amino acids and to provide evidence for the utilization of such amino acids, in addition to α-L-amino acids. β-Puromycin, a putative mimetic for β-aminoacyl-tRNAs, was used to select modified ribosome variants having altered PTC architectures, thus potentially enabling incorporation of β-L-amino acids. Eight types of modified ribosomes altered within the PTC have been selected by monitoring improved sensitivity to β-puromycin in vivo. Two of the modified ribosomes, having 2057AGCGUGA2063 and 2502UGGCAG2507 or 2502AGCCAG2507, were able to suppress UAG codons in E. coli dihydrofolate reductase (DHFR) and scorpion Opisthorcanthus madagascariensis peptide IsCT mRNAs in the presence of β-alanyl-tRNA(CUA). PMID:22145951

  3. Genetic Incorporation of Unnatural Amino Acids into Proteins in Mycobacterium tuberculosis

    Wang, Feng; Robbins, Scott; Guo, Jiantao; Shen, Weijun; Schultz, Peter G.

    2010-01-01

    New tools are needed to study the intracellular pathogen Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB), to facilitate new drug discovery and vaccine development. We have developed methodology to genetically incorporate unnatural amino acids into proteins in Mycobacterium smegmatis, BCG and Mtb, grown both extracellularly in culture and inside host cells. Orthogonal mutant tRNATyr/tyrosyl-tRNA synthetase pairs derived from Methanococcus jannaschii and evolved in Es...

  4. Enhanced incorporation of fatty acid into phosphatidyl choline that parallels histamine discharge in mast cells

    Purified rat peritoneal and pleural mast cells preincubated briefly with radioactively labeled fatty acid were treated with A23187, which bypasses primary receptors in stimulating exocytosis. An enhanced incorporation of fatty acid into phosphatidyl choline (PC) that occurred in parallel with histamine release at 24-25 degrees C was observed and was initially proportional to the total amount of histamine discharged. Enhanced PC labeling and histamine secretion were also proportional at temperatures ranging from 17-37 degrees C. Both radioactive linoleic and palmitic acids were incorporated selectively at the beta-position of the glycerol backbone of PC. PC labeling by [3H]choline was not detectably different in control and stimulated cells, and phosphatidic acid did not exhibit selectively enhanced beta-acylation. Thus, the stimulated labeling in A23187-treated cells may occur secondary to the action of a phospholipase A2 that favors PC as a substrate. Other peritoneal cell types exhibit a very similar A23187-stimulated selective labeling of PC. Therefore, autoradiography has been used to provide a direct demonstration that in purified preparations, mast cells are the principal cell type engaged in A23187-elicited incorporation of fatty acid into PC. The efficacy of this approach has relied on special procedures devised to obtain significantly different autoradiographic grain densities between control and stimulated preparations that can be attributed to differences in the level of [3H]palmitate-labeled PC. Preliminary tests using compound 48/80 as a secretory stimulus for mast cells have identified a similar selectively enhanced PC labeling. In either case, however, consideration of possible relationships between PC metabolism and the secretory process are premature since they have not been tested directly

  5. Incorporation of arachidonic acid (AA) into phosphatidylcholine molecular species of the human neutrophil (PMN)

    Recently the authors proposed that the initial incorporation of AA into 1,2 diacylphosphatidylcholine (PC) was mediated by AA-CoA transferase(s) while the subsequent transfer of AA from 1,2-diacyl- into alkyl, acyl-PC was mediated by a CoA-independent transacylase. Studies here provide further evidence for such a two-step mechanism. PMNs were pulse labeled for 5 min with 3H-AA (.07μM) which was rapidly incorporated into 1,2-diacyl-PC. However, incorporation of AA into 1,2-diacyl-PC was inhibited by incubation with high levels of AA (30 μM). Similarly PMNs were pulsed labeled with 3H-AA for 5 min followed by a 120 min incubation. In these cells, 3H-AA was rapidly transferred from 1,2-diacyl-PC into alkyl, acyl-PC. In the presence of 30 μM AA redistribution of 3H-AA from diacyl to alkyl, acyl-PC was observed. This result implied that the initial incorporation of 3H-AA proceeds via a free acid intermediate while the transfer of 3H-AA from diacyl to alkyl, acyl-PC does not. Using a cell free system, 14C-AACoA was incubated for 5 min and found to be incorporated into 1,2-diacyl-PC containing 16:0, 18:0, and 18:1 at the sn-1 position. Furthermore 14C-AACoA and various 1-radyl, 2-lyso-PC were added to a PMN membrane preparation. The arachidonyl-transferase(s) preferred the 1-acyl, 2-lyso-PC substrate to 1-alkyl, 2-lyso-PC. Thus these studies provide further evidence that AA is initially incorporated into 1,2-diacyl-PC through arachidonyl-CoA transferases

  6. Rationally evolving tRNAPyl for efficient incorporation of noncanonical amino acids.

    Fan, Chenguang; Xiong, Hai; Reynolds, Noah M; Söll, Dieter

    2015-12-15

    Genetic encoding of noncanonical amino acids (ncAAs) into proteins is a powerful approach to study protein functions. Pyrrolysyl-tRNA synthetase (PylRS), a polyspecific aminoacyl-tRNA synthetase in wide use, has facilitated incorporation of a large number of different ncAAs into proteins to date. To make this process more efficient, we rationally evolved tRNA(Pyl) to create tRNA(Pyl-opt) with six nucleotide changes. This improved tRNA was tested as substrate for wild-type PylRS as well as three characterized PylRS variants (N(ϵ)-acetyllysyl-tRNA synthetase [AcKRS], 3-iodo-phenylalanyl-tRNA synthetase [IFRS], a broad specific PylRS variant [PylRS-AA]) to incorporate ncAAs at UAG codons in super-folder green fluorescence protein (sfGFP). tRNA(Pyl-opt) facilitated a 5-fold increase in AcK incorporation into two positions of sfGFP simultaneously. In addition, AcK incorporation into two target proteins (Escherichia coli malate dehydrogenase and human histone H3) caused homogenous acetylation at multiple lysine residues in high yield. Using tRNA(Pyl-opt) with PylRS and various PylRS variants facilitated efficient incorporation of six other ncAAs into sfGFP. Kinetic analyses revealed that the mutations in tRNA(Pyl-opt) had no significant effect on the catalytic efficiency and substrate binding of PylRS enzymes. Thus tRNA(Pyl-opt) should be an excellent replacement of wild-type tRNA(Pyl) for future ncAA incorporation by PylRS enzymes. PMID:26250114

  7. Microbial release of 2,4-dichlorophenyl bound to humic acid or incorporated during humification

    The microbial release of 14C-labeled 2,4-dichlorophenol (DCP) bound to synthetic and natural humic materials or polymerized by enzymes was investigated to evaluate the possibility of future adverse effects if binding or polymerization is used for decontamination purposes. After 12 wk of incubation with microorganisms obtained from a forest soil, the amounts of substances released into the media were very small (maximum 2.2% of the initially bound 14C) without regard to the kind of the polymer into which 14C-DCP was incorporated. Most of the radioactivity (46.2 to 80.8%) remained bound to the precipitated humic materials or in the DCP-polymer (90.1 to 97.0%). Certain amounts of the released substances evolved in the form of 14CO2 (1.0 to 9.4% from humic materials and 0 to 0.5% from a DCP-polymer). The rate of mineralization differed depending on the type of DCP binding - surface-bound or incorporated during synthesis of humic acid - and on the kind of polymer to which 14C-DCP was attached. The release into the media and 14CO2 evolution for synthetic and natural humic acids was essentially the same. When only DCP was present in the growth medium, the formation of 14CO2 was less than from a DCP-humic acid complex; this may indicate that 14CO2 from a DCP-humic acid complex originated mostly from DCP derivatives. The data obtained for DCP did not provide any evidence for a delayed pollution problem associated with polymerization or binding of xenobiotics to humic acids

  8. Alpha-synuclein gene ablation increases docosahexaenoic acid incorporation and turnover in brain phospholipids

    Golovko, Mikhail Y; Rosenberger, Thad A; Feddersen, Søren;

    2007-01-01

    incorporation rate and turnover in ethanolamine glycerophospholipid, phosphatidylserine, and phosphatidylinositol pools. Increased 22:6n-3-CoA mass was not the result of altered Acsl activity, which was unaffected by the absence of Snca. While Snca bound 22:6n-3, Kd = 1.0 +/- 0.5 micromol/L, it did not bind 22......Previously, we demonstrated that ablation of alpha-synuclein (Snca) reduces arachidonate (20:4n-6) turnover in brain phospholipids through modulation of an endoplasmic reticulum-localized acyl-CoA synthetase (Acsl). The effect of Snca ablation on docosahexaenoic acid (22:6n-3) metabolism is unknown...

  9. Stable isotope studies reveal pathways for the incorporation of non-essential amino acids in Acyrthosiphon pisum (pea aphids).

    Haribal, Meena; Jander, Georg

    2015-12-01

    Plant roots incorporate inorganic nitrogen into the amino acids glutamine, glutamic acid, asparagine and aspartic acid, which together serve as the primary metabolites of nitrogen transport to other tissues. Given the preponderance of these four amino acids, phloem sap is a nutritionally unbalanced diet for phloem-feeding insects. Therefore, aphids and other phloem feeders typically rely on microbial symbionts for the synthesis of essential amino acids. To investigate the metabolism of the four main transport amino acids by the pea aphid (Acyrthosiphon pisum), and its Buchnera aphidicola endosymbionts, aphids were fed defined diets with stable isotope-labeled glutamine, glutamic acid, asparagine or aspartic acid (U-(13)C, U-(15)N; U-(15)N; α-(15)N; or γ-(15)N). The metabolic fate of the dietary (15)N and (13)C was traced using gas chromatography-mass spectrometry (GC-MS). Nitrogen was the major contributor to the observed amino acid isotopomers with one additional unit mass (M+1). However, there was differential incorporation, with the amine nitrogen of asparagine being incorporated into other amino acids more efficiently than the amide nitrogen. Higher isotopomers (M+2, M+3 and M+4) indicated the incorporation of varying numbers of (13)C atoms into essential amino acids. GC-MS assays also showed that, even with an excess of dietary labeled glutamine, glutamic acid, asparagine or aspartic acid, the overall content of these amino acids in aphid bodies was mostly the product of catabolism of dietary amino acids and subsequent re-synthesis within the aphids. Thus, these predominant dietary amino acids are not passed directly to Buchnera endosymbionts for synthesis of essential amino acids, but are rather are produced de novo, most likely by endogenous aphid enzymes. PMID:26632455

  10. Targeting lipopolyplexes using bifunctional peptides incorporating hydrophobic spacer amino acids: synthesis, transfection, and biophysical studies.

    Pilkington-Miksa, Michael A; Writer, Michele J; Sarkar, Supti; Meng, Qing-Hai; Barker, Suzie E; Shamlou, Parviz Ayazi; Hailes, Helen C; Hart, Stephen L; Tabor, Alethea B

    2007-01-01

    We have developed efficient synthetic routes to two hydrophobic amino acids, suitably protected for solid-phase peptide synthesis, and have successfully synthesized peptides containing these or other hydrophobic amino acids as spacers between a Lys16 moiety and an integrin-targeting motif. These peptides have in turn been used to formulate a range of lipopolyplex vectors with Lipofectin and plasmid DNA. The transfection efficiencies of these vectors and their aggregation behavior in buffers and in serum have been studied. We have shown that vectors containing peptides incorporating long linkers that are entirely hydrophobic are less efficient transfection agents. However, linkers of equivalent length that are in part hydrophobic show improved transfection properties, which is probably due to the improved accessibility of the integrin-binding motif. PMID:17915956

  11. Starch-based Antimicrobial Films Incorporated with Lauric Acid and Chitosan

    Salleh, E.; Muhamad, I. I.

    2010-03-01

    Antimicrobial (AM) packaging is one of the most promising active packaging systems. Starch-based film is considered an economical material for antimicrobial packaging. This study aimed at the development of food packaging based on wheat starch incorporated with lauric acid and chitosan as antimicrobial agents. The purpose is to restrain or inhibit the growth of spoilage and/or pathogenic microorganisms that are contaminating foods. The antimicrobial effect was tested on B. substilis and E. coli. Inhibition of bacterial growth was examined using two methods, i.e. zone of inhibition test on solid media and liquid culture test (optical density measurements). The control and AM films (incorporated with chitosan and lauric acid) were produced by casting method. From the observations, AM films exhibited inhibitory zones. Interestingly, a wide clear zone on solid media was observed for B. substilis growth inhibition whereas inhibition for E. coli was not as effective as B. substilis. From the liquid culture test, the AM films clearly demonstrated a better inhibition against B. substilis than E. coli.

  12. Incorporation of non-canonical amino acids into the developing murine proteome.

    Calve, Sarah; Witten, Andrew J; Ocken, Alexander R; Kinzer-Ursem, Tamara L

    2016-01-01

    Analysis of the developing proteome has been complicated by a lack of tools that can be easily employed to label and identify newly synthesized proteins within complex biological mixtures. Here, we demonstrate that the methionine analogs azidohomoalanine and homopropargylglycine can be globally incorporated into the proteome of mice through facile intraperitoneal injections. These analogs contain bio-orthogonal chemical handles to which fluorescent tags can be conjugated to identify newly synthesized proteins. We show these non-canonical amino acids are incorporated into various tissues in juvenile mice and in a concentration dependent manner. Furthermore, administration of these methionine analogs to pregnant dams during a critical stage of murine development, E10.5-12.5 when many tissues are assembling, does not overtly disrupt development as assessed by proteomic analysis and normal parturition and growth of pups. This successful demonstration that non-canonical amino acids can be directly administered in vivo will enable future studies that seek to characterize the murine proteome during growth, disease and repair. PMID:27572480

  13. Antifouling polyethersulfone hemodialysis membranes incorporated with poly (citric acid) polymerized multi-walled carbon nanotubes.

    Abidin, Muhammad Nidzhom Zainol; Goh, Pei Sean; Ismail, Ahmad Fauzi; Othman, Mohd Hafiz Dzarfan; Hasbullah, Hasrinah; Said, Noresah; Kadir, Siti Hamimah Sheikh Abdul; Kamal, Fatmawati; Abdullah, Mohd Sohaimi; Ng, Be Cheer

    2016-11-01

    Poly (citric acid)-grafted-MWCNT (PCA-g-MWCNT) was incorporated as nanofiller in polyethersulfone (PES) to produce hemodialysis mixed matrix membrane (MMM). Citric acid monohydrate was polymerized onto the surface of MWCNTs by polycondensation. Neat PES membrane and PES/MWCNTs MMMs were fabricated by dry-wet spinning technique. The membranes were characterized in terms of morphology, pure water flux (PWF) and bovine serum albumin (BSA) protein rejection. The grafting yield of PCA onto MWCNTs was calculated as 149.2%. The decrease of contact angle from 77.56° to 56.06° for PES/PCA-g-MWCNTs membrane indicated the increase in surface hydrophilicity, which rendered positive impacts on the PWF and BSA rejection of the membrane. The PWF increased from 15.8Lm(-2)h(-1) to 95.36Lm(-2)h(-1) upon the incorporation of PCA-g-MWCNTs due to the attachment of abundant hydrophilic groups that present on the MWCNTs, which have improved the affinity of membrane towards the water molecules. For protein rejection, the PES/PCA-g-MWCNTs MMM rejected 95.2% of BSA whereas neat PES membrane demonstrated protein rejection of 90.2%. Compared to commercial PES hemodialysis membrane, the PES/PCA-g-MWCNTs MMMs showed less flux decline behavior and better PWF recovery ratio, suggesting that the membrane antifouling performance was improved. The incorporation of PCA-g-MWCNTs enhanced the separation features and antifouling capabilities of the PES membrane for hemodialysis application. PMID:27524052

  14. Red-Shifted Aequorin Variants Incorporating Non-Canonical Amino Acids: Applications in In Vivo Imaging

    Grinstead, Kristen M.; Rowe, Laura; Ensor, Charles M.; Joel, Smita; Daftarian, Pirouz; Dikici, Emre; Zingg, Jean-Marc; Daunert, Sylvia

    2016-01-01

    The increased importance of in vivo diagnostics has posed new demands for imaging technologies. In that regard, there is a need for imaging molecules capable of expanding the applications of current state-of-the-art imaging in vivo diagnostics. To that end, there is a desire for new reporter molecules capable of providing strong signals, are non-toxic, and can be tailored to diagnose or monitor the progression of a number of diseases. Aequorin is a non-toxic photoprotein that can be used as a sensitive marker for bioluminescence in vivo imaging. The sensitivity of aequorin is due to the fact that bioluminescence is a rare phenomenon in nature and, therefore, it does not suffer from autofluorescence, which contributes to background emission. Emission of bioluminescence in the blue-region of the spectrum by aequorin only occurs when calcium, and its luciferin coelenterazine, are bound to the protein and trigger a biochemical reaction that results in light generation. It is this reaction that endows aequorin with unique characteristics, making it ideally suited for a number of applications in bioanalysis and imaging. Herein we report the site-specific incorporation of non-canonical or non-natural amino acids and several coelenterazine analogues, resulting in a catalog of 72 cysteine-free, aequorin variants which expand the potential applications of these photoproteins by providing several red-shifted mutants better suited to use in vivo. In vivo studies in mouse models using the transparent tissue of the eye confirmed the activity of the aequorin variants incorporating L-4-iodophehylalanine and L-4-methoxyphenylalanine after injection into the eye and topical addition of coelenterazine. The signal also remained localized within the eye. This is the first time that aequorin variants incorporating non-canonical amino acids have shown to be active in vivo and useful as reporters in bioluminescence imaging. PMID:27367859

  15. Red-Shifted Aequorin Variants Incorporating Non-Canonical Amino Acids: Applications in In Vivo Imaging.

    Kristen M Grinstead

    Full Text Available The increased importance of in vivo diagnostics has posed new demands for imaging technologies. In that regard, there is a need for imaging molecules capable of expanding the applications of current state-of-the-art imaging in vivo diagnostics. To that end, there is a desire for new reporter molecules capable of providing strong signals, are non-toxic, and can be tailored to diagnose or monitor the progression of a number of diseases. Aequorin is a non-toxic photoprotein that can be used as a sensitive marker for bioluminescence in vivo imaging. The sensitivity of aequorin is due to the fact that bioluminescence is a rare phenomenon in nature and, therefore, it does not suffer from autofluorescence, which contributes to background emission. Emission of bioluminescence in the blue-region of the spectrum by aequorin only occurs when calcium, and its luciferin coelenterazine, are bound to the protein and trigger a biochemical reaction that results in light generation. It is this reaction that endows aequorin with unique characteristics, making it ideally suited for a number of applications in bioanalysis and imaging. Herein we report the site-specific incorporation of non-canonical or non-natural amino acids and several coelenterazine analogues, resulting in a catalog of 72 cysteine-free, aequorin variants which expand the potential applications of these photoproteins by providing several red-shifted mutants better suited to use in vivo. In vivo studies in mouse models using the transparent tissue of the eye confirmed the activity of the aequorin variants incorporating L-4-iodophehylalanine and L-4-methoxyphenylalanine after injection into the eye and topical addition of coelenterazine. The signal also remained localized within the eye. This is the first time that aequorin variants incorporating non-canonical amino acids have shown to be active in vivo and useful as reporters in bioluminescence imaging.

  16. Improved Incorporation of Noncanonical Amino Acids by an Engineered tRNA(Tyr) Suppressor.

    Rauch, Benjamin J; Porter, Joseph J; Mehl, Ryan A; Perona, John J

    2016-01-26

    The Methanocaldcoccus jannaschii tyrosyl-tRNA synthetase (TyrRS):tRNA(Tyr) cognate pair has been used to incorporate a large number of noncanonical amino acids (ncAAs) into recombinant proteins in Escherichia coli. However, the structural elements of the suppressor tRNA(Tyr) used in these experiments have not been examined for optimal performance. Here, we evaluate the steady-state kinetic parameters of wild-type M. jannaschii TyrRS and an evolved 3-nitrotyrosyl-tRNA synthetase (nitroTyrRS) toward several engineered tRNA(Tyr) suppressors, and we correlate aminoacylation properties with the efficiency and fidelity of superfolder green fluorescent protein (sfGFP) synthesis in vivo. Optimal ncAA-sfGFP synthesis correlates with improved aminoacylation kinetics for a tRNA(Tyr) amber suppressor with two substitutions in the anticodon loop (G34C/G37A), while four additional mutations in the D and variable loops, present in the tRNA(Tyr) used in all directed evolution experiments to date, are deleterious to function both in vivo and in vitro. These findings extend to three of four other evolved TyrRS enzymes that incorporate distinct ncAAs. Suppressor tRNAs elicit decreases in amino acid Km values for both TyrRS and nitroTyrRS, suggesting that direct anticodon recognition by TyrRS need not be an impediment to superior performance of this orthogonal system and offering insight into novel approaches for directed evolution. The G34C/G37A tRNA(Tyr) may enhance future incorporation of many ncAAs by engineered TyrRS enzymes. PMID:26694948

  17. Antibacterial Studies and Effect of Poloxamer on Gold Nanoparticles by Zingiber Officinale Extracted Green Synthesis.

    Chitra, K; Reena, K; Manikandan, A; Antony, S Arul

    2015-07-01

    Poloxamer finds excellent clinical and therapeutic uses for curing of various ailments. The Zin- giber officinale (Z. officinale) is one of the well-known medicinal plants. The poloxamer188 and the rhizome extract of Z. officinale have been used to synthesize the gold nanoparticles (AuNPs) by a green approach. The Z. officinale extract has been used as a reducing agent while the polox- amerl88 has been used as a stabilizing agent. The effect of addition of poloxamer on the controlling the shape and size of the AuNPs has been investigated by transmission electron microscopy (TEM) and dynamic light scattering techniques. The formation of AuNPs has also been confirmed by UV-Visible spectral, energy dispersive X-ray (EDX) and powder X-ray diffraction (XRD) analyses. The anti-bacterial activity of the green synthesized AuNPs has been investigated on the three human pathogens Staphylococcus aureus, Escherichia coli, and Klebsiella pneumonia. The poloxamer188 protected AuNPs inhibit the bacterial growth more effectively than the pure Z. officinale extract and the standard tetracycline (TA). PMID:26373065

  18. A FACTORIAL STUDY ON THE EFFECTS OF HYDROXYPROPYL β CYCLODEXTRIN AND POLOXAMER ON THE SOLUBILITY AND DISSOLUTION RATE OF NIMESULIDE

    K.P.R. Chowdary et al.

    2012-01-01

    The objective of the study is to evaluate the individual and combined effects of hydroxy propyl β cyclodextrin (HPβCD), surfactant (Poloxamer 407) on the solubility and dissolution rate of nimesulide, a BCS class II drug in a series of 22 factorial experiments. The solubility of nimesulide in four selected fluids containing HPβCD and Poloxamer 407 as per a 22 factorial study was determined. HPβCD alone gave a marginal increase (1.47 fold) in the solubility of nimesulide. Whereas Poloxamer 407...

  19. Evaluating antibiotics for use in medicine using a poloxamer biofilm model

    Cochrane Christine A

    2007-02-01

    Full Text Available Abstract Background Wound infections, due to biofilms, are a constant problem because of their recalcitrant nature towards antibiotics. Appropriate antibiotic selection for the treatment of these biofilm infections is important. The traditional in vitro disc diffusion method for antibiotic selection uses bacterial cultures grown on agar plates. However, the form of bacterial growth on agar is not representative of how bacteria grow in wounds and other tissue sites as here bacteria grow naturally in a biofilm. The aim of this research was to test a more appropriate method for testing antimicrobial efficacy on biofilms and compare with the standard methods used for antibiotic sensitivity testing. Methods Outer Membrane Protein analysis was performed on E.coli, Staphylococcus aureus, Pseudomonas aeruginosa, Proteus mirabilis and Acinetobacter juni when grown on Mueller Hinton agar ('quasi-biofilm state' and 30% Poloxamer hydrogel ('true- biofilm state. Susceptibility to antibiotics on 28 clinical isolates was determined using the modified Kirby Bauer disc diffusion method, on agar and 30% Poloxamer. Results Similar outer membrane proteins [OMPs] were identified in bacteria grown in a biofilm state and on a 30% poloxamer hydrogel, which were very different to the OMPs identified in bacteria grown on Mueller-Hinton agar and broth. There was a significant difference between the means of the clearance zones around the antibiotic discs on standard agar and poloxamer gels [P 0.05]. Conclusion The findings of this experiment suggest that poloxamer gel could be used as an appropriate medium on which to conduct biofilm antibiotic susceptibility tests as it enables bacteria to be grown in a state representative of the infected surface from which the culture was taken.

  20. Cyclodextrin-poloxamer aggregates as nanocarriers in eye drop formulations: dexamethasone and amphotericin B.

    Jansook, Phatsawee; Pichayakorn, Wiwat; Muankaew, Chutimon; Loftsson, Thorsteinn

    2016-09-01

    In this present study cyclodextrin (CD)-poloxamer aggregates were characterized and developed as ophthalmic drug carriers. The combined effect of γCD/2-hydroxypropyl-γCD (HPγCD) mixtures and poloxamer on solubilization and permeability of two model drugs, dexamethasone (Dex) and amphotericin B (AmB), was investigated. The CD-poloxamer interaction and complex aggregation were examined by (1)H nuclear magnetic resonance ((1)H-NMR), their solubilizing ability by high-performance liquid chromatography, and their particle size determined by dynamic light scattering and transmission electron microscopy. Formulations containing either 1.5% w/v Dex or 0.15% w/v AmB in eye drop suspensions containing various γCD/HPγCD ratios and poloxamer 407 (P407) were prepared. The solubility of the drugs, surface tension and hemolytic effect of the eye drops and drug permeation from selected formulations were determined. The (1)H-NMR study showed that P407 formed inclusion complex with CDs by inserting its poly(propylene oxide) segment into the CD cavity. P407 and γCD interacted with each other to form nanosized aggregates, and the observed concentration of dissolved γCD and P407 progressively decreased with increasing γCD and P407 concentrations. Including a high proportion of HPγCD improved the drug solubilization and reduced the hemolytic effect. The surface tension of the formulations decreased with increasing P407 concentration. Furthermore, increasing P407 content in the formulations enhanced formation of complex aggregates with consequent slower drug release. It was concluded that the drug/γCD/HPγCD complex was stabilized by P407 through formation of multi-component aggregates. Thus, CD-poloxamer aggregates are self-assembled nanocarriers from which drug delivery characteristics can be adjusted by changing the γCD/HPγCD/P407 ratios. PMID:26765786

  1. Direct incorporation of fatty acids into microbial phospholipids in soils: Position-specific labeling tells the story

    Dippold, Michaela A.; Kuzyakov, Yakov

    2016-02-01

    Fatty acids have been used as plant and microbial biomarkers, and knowledge about their transformation pathways in soils and sediments is crucial for interpreting fatty acid signatures, especially because the formation, recycling and decomposition processes are concurrent. We analyzed the incorporation of free fatty acids into microbial fatty acids in soil by coupling position-specific 13C labeling with compound-specific 13C analysis. Position-specifically and uniformly 13C labeled palmitate were applied in an agricultural Luvisol. Pathways of fatty acids were traced by analyzing microbial utilization of 13C from individual molecule positions of palmitate and their incorporation into phospholipid fatty acids (PLFA). The fate of palmitate 13C in the soil was characterized by the main pathways of microbial fatty acid metabolism: Odd positions (C-1) were preferentially oxidized to CO2 in the citric acid cycle, whereas even positions (C-2) were preferentially incorporated into microbial biomass. This pattern is a result of palmitate cleavage to acetyl-CoA and its further use in the main pathways of C metabolism. We observed a direct, intact incorporation of more than 4% of the added palmitate into the PLFA of microbial cell membranes, indicating the important role of palmitate as direct precursor for microbial fatty acids. Palmitate 13C was incorporated into PLFA as intact alkyl chain, i.e. the C backbone of palmitate was not cleaved, but palmitate was incorporated either intact or modified (e.g. desaturated, elongated or branched) according to the fatty acid demand of the microbial community. These modifications of the incorporated palmitate increased with time. Future PLFA studies must therefore consider the recycling of existing plant and microbial-derived fatty acids. This study demonstrates the intact uptake and recycling of free fatty acids such as palmitate in soils, as well as the high turnover and transformation of cellular PLFA. Knowledge about the intact

  2. Tunable translational control using site-specific unnatural amino acid incorporation in Escherichia coli

    Yusuke Kato

    2015-04-01

    Full Text Available Translation of target gene transcripts in Escherichia coli harboring UAG amber stop codons can be switched on by the amber-codon-specific incorporation of an exogenously supplied unnatural amino acid, 3-iodo-L-tyrosine. Here, we report that this translational switch can control the translational efficiency at any intermediate magnitude by adjustment of the 3-iodo-L-tyrosine concentration in the medium, as a tunable translational controller. The translational efficiency of a target gene reached maximum levels with 10−5 M 3-iodo-L-tyrosine, and intermediate levels were observed with suboptimal concentrations (approximately spanning a 2-log10 concentration range, 10−7–10−5 M. Such intermediate-level expression was also confirmed in individual bacteria.

  3. Effective removal of humic acid using xanthan gum incorporated polyethersulfone membranes.

    Sathish Kumar, R; Arthanareeswaran, G; Paul, Diby; Kweon, Ji Hyang

    2015-11-01

    In this study, xanthan gum (XA) was used as a hydrophilic biopolymer additive for the modification of polyethersulfone (PES) membrane to removal of humic acid (HA). The membranes are prepared using phase inversion technique and the concentration of XA was varied from 0.5 to 1.5wt%. The prepared membranes are characterized as a function of hydrophilicity, equilibrium water content (EWC), porosity studies and functional group analysis. Membrane surface and cross-sectional morphology was studied using scanning electron microscope. The lower contact angle value 64.2° was exhibited, when 1.5wt% of XA incorporated in PES membrane and this ensures that increase of hydrophilicity in pristine PES membrane. Further, higher water permeability (PWP) of 68.9(-9)m/skPa was observed for 1.5wt% of XA/PES membrane. The effect of pH on HA removal was studied for neat PES and XA/PES membranes. The rejection performance of XA incorporated in PES membranes were compared with commercial available PES membrane. PMID:25857244

  4. Effect of polyunsaturated fatty acids and phospholipids on [3H]-vitamin E incorporation into pulmonary artery endothelial cell membranes

    Vitamin E, a dietary antioxidant, is presumed to be incorporated into the lipid bilayer of biological membranes to an extent proportional to the amount of polyunsaturated fatty acids or phospholipids in the membrane. In the present study we evaluated the distribution of incorporated polyunsaturated fatty acids (PUFA) and phosphatidylethanolamine (PE) in various membranes of pulmonary artery endothelial cells. We also studied whether incorporation of PUFA or PE is responsible for increased incorporation of [3H]-vitamin E into the membranes of these cells. Following a 24-hr incubation with linoleic acid (18:2), 18:2 was increased by 6.9-, 9.2-, and 13.2-fold in plasma, mitochondrial, and microsomal membranes, respectively. Incorporation of 18:2 caused significant increases in the unsaturation indexes of mitochondrial and microsomal polyunsaturated fatty acyl chains (P less than .01 versus control in both membranes). Incubation with arachidonic acid (20:4) for 24 hr resulted in 1.5-, 2.3-, and 2.4-fold increases in 20:4 in plasma, mitochondrial, and microsomal membranes, respectively. The unsaturation indexes of polyunsaturated fatty acyl chains of mitochondrial and microsomal membranes also increased (P less than .01 versus control in both membranes). Although incubations with 18:2 or 20:4 resulted in several-fold increases in membrane 18:2 or 20:4 fatty acids, incorporation of [3H]-vitamin E into these membranes was similar to that in controls. Following a 24-hr incubation with PE, membrane PE content was significantly increased, and [3H]-vitamin E incorporation was also increased to a comparable degree, i.e., plasma membrane greater than mitochondria greater than microsomes. Endogenous vitamin E content of the cells was not altered because of increased incorporation of PE and [3H]-vitamin E

  5. Comparative evaluation on fatty acid and Matricaria recutita essential oil incorporated into casein-based film.

    Aliheidari, Nahal; Fazaeli, Mahboubeh; Ahmadi, Reza; Ghasemlou, Mehran; Emam-Djomeh, Zahra

    2013-05-01

    Sodium caseinate composite films containing lipids-oleic acid (OA), stearic acid (SA), or Matricaria recutita essential oil (MEO) - were prepared through emulsification and their physical, thermal, mechanical, and barrier properties were evaluated and compared. Furthermore, their antimicrobial effectiveness against Listeria monocytogenes, Staphylococcus aureus, and Escherichia coli was studied. Emulsified films were softer, less rigid, and more stretchable than pure films. The films' water vapor barrier properties were found to decrease upon the addition of lipid content; this effect was greatly reduced when MEO was added. The presence of OA/SA and MEO decreased tensile strength and elastic modulus but increased the elongation at break. Thermal analysis of all emulsified films showed two endothermic peaks; these results confirmed those obtained by SEM studies, where a partial separation of the two phases occurred. The films' antimicrobial activities were increased by incorporating lipids, particularly those containing MEO, which were more effective against the studied bacteria. This work showed that when taking all the studied variables into account, films formulated with MEO were found most suitable for various food applications. PMID:23415659

  6. Physicochemical Properties of Edible Chitosan/Hydroxypropyl Methylcellulose/Lysozyme Films Incorporated with Acidic Electrolyzed Water

    Ewa Brychcy

    2015-01-01

    Full Text Available The treatment with acidic electrolyzed water (AEW is a promising disinfection method due to its effectiveness in reducing microbial population. The aim of the study was to evaluate physicochemical properties of chitosan/HPMC films incorporated with lysozyme and acidic electrolyzed water. In the composite films, decreasing film solubility and increasing concentration of sodium chloride solution and prolongation of electrolysis time were observed. Electrolysis process with sodium chloride induces spongy network of film structure. The use of AEW has not changed chemical composition of films which was proved by 1H NMR, MALDI-TOF, and FT-IR spectroscopy. The research confirmed that electrolysis significantly improved thermomechanical properties of the examined films. The contact angle values of the films were quite similar and ranged between 56° and 73°. The increase of salt concentration used in the electrolysis process had an impact on increasing flexibility of samples. Application of electrolyzed water in commonly used food processing systems is possible. Fusion of AEW and biopolymers may provide better integration with coated food product and multidirectional protecting effect.

  7. Palmitic acid-labeled lipids selectively incorporated into platelet cytoskeleton during aggregation

    Previous experiments showed that during the early stages (20-30 seconds) of aggregation induced by adenosine diphosphate (ADP, 2 microM) or thrombin (0.1 U/mL) of rabbit or human platelets prelabeled with [3H]palmitic acid, labeled lipid became associated with the cytoskeleton isolated after lysis with 1% Triton X-100, 5 mM EGTA [ethylene glycol-bis-(beta-aminoethyl ether)]-N,N,N',N'-tetra-acetic acid. The association appeared to be related to the number of sites of contact and was independent of the release of granule contents. We have now investigated the nature of the labeled lipids by thin-layer and column chromatography and found differences between the distribution of the label in intact platelets (both stimulated and unstimulated) and the isolated cytoskeletons. In both species, and with either ADP or thrombin as aggregating agent, 70-85% of the label in both intact platelets and in the cytoskeletons was in phospholipids. The distribution of label among the phospholipids in the cytoskeletons was similar to that in intact platelets except that the percentage of label in phosphatidylcholine was significantly higher in the cytoskeletons of human platelets than in the intact platelets, and the percentage of label in phosphatidylserine/phosphatidylinositol was significantly lower in the cytoskeletons of rabbit platelets and thrombin-aggregated human platelets than in intact platelets. The cytoskeletons contained a lower percentage of label in triacylglycerol, diacylglycerol, and cholesterol ester than the intact platelets. Contrary to a report in the literature, we found no evidence for the incorporation of diacylglycerol and palmitic acid into the cytoskeleton

  8. Type 1 diacylglycerol acyltransferases of Brassica napus preferentially incorporate oleic acid into triacylglycerol

    Aznar-Moreno, Jose; Denolf, Peter; Van Audenhove, Katrien; De Bodt, Stefanie; Engelen, Steven; Fahy, Deirdre; Wallis, James G.; Browse, John

    2015-01-01

    DGAT1 enzymes (acyl-CoA:diacylglycerol acyltransferase 1, EC 2.3.1.20) catalyse the formation of triacylglycerols (TAGs), the most abundant lipids in vegetable oils. Thorough understanding of the enzymology of oil accumulation is critical to the goal of modifying oilseeds for improved vegetable oil production. Four isoforms of BnDGAT1, the final and rate-limiting step in triacylglycerol synthesis, were characterized from Brassica napus, one of the world’s most important oilseed crops. Transcriptional profiling of developing B. napus seeds indicated two genes, BnDGAT1-1 and BnDGAT1-2, with high expression and two, BnDGAT1-3 and BnDGAT1-4, with low expression. The activities of each BnDGAT1 isozyme were characterized following expression in a strain of yeast deficient in TAG synthesis. TAG from B. napus seeds contain only 10% palmitic acid (16:0) at the sn-3 position, so it was surprising that all four BnDGAT1 isozymes exhibited strong (4- to 7-fold) specificity for 16:0 over oleic acid (18:1) as the acyl-CoA substrate. However, the ratio of 18:1-CoA to 16:0-CoA in B. napus seeds during the peak period of TAG synthesis is 3:1. When substrate selectivity assays were conducted with 18:1-CoA and 16:0-CoA in a 3:1 ratio, the four isozymes incorporated 18:1 in amounts 2- to 5-fold higher than 16:0. This strong sensitivity of the BnDGAT1 isozymes to the relative concentrations of acyl-CoA substrates substantially explains the observed fatty acid composition of B. napus seed oil. Understanding these enzymes that are critical for triacylglycerol synthesis will facilitate genetic and biotechnological manipulations to improve this oilseed crop. PMID:26195728

  9. Characterization of Active Packaging Films Made from Poly(Lactic Acid)/Poly(Trimethylene Carbonate) Incorporated with Oregano Essential Oil

    Dong Liu; Hongli Li; Lin Jiang; Yongming Chuan; Minglong Yuan; Haiyun Chen

    2016-01-01

    Antimicromial and antioxidant bioactive films based on poly(lactic acid)/poly(trimenthylene carbonate) films incorporated with different concentrations of oregano essential oil (OEO) were prepared by solvent casting. The antimicrobial, antioxidant, physical, thermal, microstructural, and mechanical properties of the resulting films were examined. Scanning electron microscopy analysis revealed that the cross-section of films became rougher when OEO was incorporated into PLA/PTMC blends. Differ...

  10. Stabilization of a human recombinant factor VIII by poloxamer 188 in relation to polysorbate 80.

    Clark, Jakson; Montgomery, Jade; Squires, Ryan; McGuire, Joseph

    2016-03-01

    Detection of enhanced surface tension depression by surfactant in the presence of protein was recently suggested as a basis for determining whether protein stabilization by that surfactant is owing to surfactant forming a steric barrier at interfaces or surfactant association with the protein. In particular, protein interaction with surfactant aggregates may lead to an increased concentration of monomers thus enhancing surfactant adsorption, or to formation of surfactant-protein complexes having little or no effect on adsorption. We compared the initial rates of surface tension depression by poloxamer 188 and polysorbate 80 (PS 80) in the presence and absence of a human recombinant factor VIII (rFVIII). Indirect evidence had suggested poloxamer 188 enters into stable associations with rFVIII in solution but does not form a steric barrier at the interface, while PS 80 behaves in contrary fashion. In this study, we show the presence of rFVIII caused an increase in the rate (reduction in the activation energy) of PS 80 adsorption, while no such change was recorded in the case of poloxamer 188. Thus, we provide substantiation for detection of protein-mediated acceleration of surfactant adsorption as a means to compare different surfactants in relation to their favored mechanism for protein stabilization. PMID:25471699

  11. The natural non-protein amino acid N-β-methylamino-L-alanine (BMAA) is incorporated into protein during synthesis.

    Glover, W Broc; Mash, Deborah C; Murch, Susan J

    2014-11-01

    N-β-methylamino-L-alanine (BMAA) is an amino acid produced by cyanobacteria and accumulated through trophic levels in the environment and natural food webs. Human exposure to BMAA has been linked to progressive neurodegenerative diseases, potentially due to incorporation of BMAA into protein. The insertion of BMAA and other non-protein amino acids into proteins may trigger protein misfunction, misfolding and/or aggregation. However, the specific mechanism by which BMAA is associated with proteins remained unidentified. Such studies are challenging because of the complexity of biological systems and samples. A cell-free in vitro protein synthesis system offers an excellent approach for investigation of changing amino acid composition in protein. In this study, we report that BMAA incorporates into protein as an error in synthesis when a template DNA sequence is used. Bicinchoninic acid assay of total protein synthesis determined that BMAA effectively substituted for alanine and serine in protein product. LC-MS/MS confirmed that BMAA was selectively inserted into proteins in place of other amino acids, but isomers N-(2-aminoethyl)glycine (AEG) and 2,4-diaminobutyric acid (DAB) did not share this characteristic. Incorporation of BMAA into proteins was significantly higher when genomic DNA from post-mortem brain was the template. About half of BMAA in the synthetic proteins was released with denaturation with sodium dodecylsulfonate and dithiothreitol, but the remaining BMAA could only be released by acid hydrolysis. Together these data demonstrate that BMAA is incorporated into the amino acid backbone of proteins during synthesis and also associated with proteins through non-covalent bonding. PMID:25096519

  12. An efficient protocol for incorporation of an unnatural amino acid in perdeuterated recombinant proteins using glucose-based media

    Venditti, Vincenzo; Fawzi, Nicolas L.; Clore, G. Marius, E-mail: mariusc@mail.nih.gov [National Institutes of Health, Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases (United States)

    2012-03-15

    The in vivo incorporation of unnatural amino acids into proteins is a well-established technique requiring an orthogonal tRNA/aminoacyl-tRNA synthetase pair specific for the unnatural amino acid that is incorporated at a position encoded by a TAG amber codon. Although this technology provides unique opportunities to engineer protein structures, poor protein yields are usually obtained in deuterated media, hampering its application in the protein NMR field. Here, we describe a novel protocol for incorporating unnatural amino acids into fully deuterated proteins using glucose-based media (which are relevant to the production, for example, of amino acid-specific methyl-labeled proteins used in the study of large molecular weight systems). The method consists of pre-induction of the pEVOL plasmid encoding the tRNA/aminoacyl-tRNA synthetase pair in a rich, H{sub 2}O-based medium prior to exchanging the culture into a D{sub 2}O-based medium. Our protocol results in high level of isotopic incorporation ({approx}95%) and retains the high expression level of the target protein observed in Luria-Bertani medium.

  13. Measurement of the incorporation of orally administered arachidonic acid into tissue lipids

    The applicability of a stable isotope method to monitor the mixing of dietary arachidonic acid with endogenous arachidonic acid in tissue lipids was evaluated. Rats were fed octadeuterated arachidonic acid during a 20-day period, and the entry of the dietary acid into lipid esters of various tissues was examined by gas chromatography-mass spectrometric (GC-MS) analysis of their fatty acids. The rats were maintained on a fat-free diet from weaning until 63 days old to enhance the ratio of the dietary acid to endogenous arachidonate. Three separate forms of eicosatetraenoic acid in the tissue lipids could be distinguished by GC-MS: octadeuterated arachidonic acid (recent dietary origin), unlabeled arachidonic acid (maternal origin) and unlabeled 4,7,10,13-eicosatetraenoic acid (originating from palmitoleic acid). The total eicosatetraenoic acid in the tissue lipids contained about 90% arachidonate from recent dietary origin in lung, kidney, heart and fat, 70% in muscle and liver and 27% in brain. The n-7 isomer of eicosatetraenoic acid was estimated to make up 6% or less of the total eicosatetraenoic acid in lung, kidney, brain, muscle and heart tissue lipids, but it comprised around 15% of the total eicosatetraenoic acid in liver. The unlabeled arachidonic acid of maternal origin thus comprised only about 10% of the eicosatetraenoic acid in all tissues examined except muscle and brain, where it was 24% and 70% of the eicosatetraenoic acid, respectively

  14. Amino acid-incorporated polymer network by thiol-ene polymerization

    R. Yokose

    2015-08-01

    Full Text Available Triallyl L-alanine (A3A and triallyl L-phenylalanine (A3F were synthesized by reactions of L-alanine and L-phenylalanine with allyl bromide in the presence of sodium hydroxide, respectively. Thiol-ene thermal polymerization of A3A or A3F with pentaerythritol-based primary tetrathiol (pS4P or pentaerythritol-based secondary tetrathiol (S4P at allyl/SH 1/1 in the presence of 2,2'-azobis(isobutyronitrile produced an amino acid-incorporated polymer network (A3ApS4P, A3A-S4P or A3F-S4P. Although the thermally cured resins were homogeneous and flat films, the corresponding thiol-ene photopolymerization did not give a successful result. Degree of swelling for each thermally cured film in N,Ndimethylformamide was much higher than that in water. The glass transition and 5% weight loss temperatures (Tg and T5 of A3F-pS4P and A3F-S4P were higher than those of A3A-pS4P and A3A-S4P, respectively. Also, A3F-pS4P and A3F-S4P exhibited much higher tensile strengths and moduli than A3A-pS4P and A3A-S4P did, respectively. Consequently, A3FpS4P displayed the highest Tg (38.7°C, T5 (282.0°C, tensile strength (9.5 MPa and modulus (406 MPa among all the thermally cured resins.

  15. Changes of free amino acids during rumen fermentation and incorporation of urea-15N into microorganisms in vitro

    Nitrogen-15 in urea was incorporated into NH3-N, bacteria-N, and 2,000 rpm-precipitate fraction-N with time course changes. The degree of incorporation was more intensely in U5 experiment than in U3 experiment. The highest 15N concentration in the bacterial fraction appeared after 6 and 12 hours after the beginning of incubation in the case of U3 and U5, respectively. The time course changes in 15N concentration in the 2,000 rpm-precipitate fraction were in parallel with that in the bacterial fraction except for 24 hours. The changes in concentration of free amino acids in the medium were observed in lysine, aspartic acid, and alanine remarkably. The concentration of total free amino acids changed with the lapse of time. The total concentration in the case of U3 experiment decreased gradually until 6 hours, and then increased until 24 hours. On the other hand, the total concentration in the case of U5 decreased continuously except for the slight increase at 6 hours. Each amino acid was isolated from the hydrolyzate of the 2,000 rpm-precipitate fraction by column chromatography. Each 15N concentration was determined for isolated amino acids. Consequently, 15N concentrations were relatively high in glysine, alanine, glutamic acid, and aspartic acid. On the contrary, 15N concentrations were very low in methionine and cystine. (JPN)

  16. Paramagnetic relaxation enhancement of membrane proteins by incorporation of the metal-chelating unnatural amino acid 2-amino-3-(8-hydroxyquinolin-3-yl)propanoic acid (HQA)

    Park, Sang Ho; Wang, Vivian S.; Radoicic, Jasmina; Angelis, Anna A. De; Berkamp, Sabrina; Opella, Stanley J., E-mail: sopella@ucsd.edu [University of California, San Diego, Department of Chemistry and Biochemistry (United States)

    2015-04-15

    The use of paramagnetic constraints in protein NMR is an active area of research because of the benefits of long-range distance measurements (>10 Å). One of the main issues in successful execution is the incorporation of a paramagnetic metal ion into diamagnetic proteins. The most common metal ion tags are relatively long aliphatic chains attached to the side chain of a selected cysteine residue with a chelating group at the end where it can undergo substantial internal motions, decreasing the accuracy of the method. An attractive alternative approach is to incorporate an unnatural amino acid that binds metal ions at a specific site on the protein using the methods of molecular biology. Here we describe the successful incorporation of the unnatural amino acid 2-amino-3-(8-hydroxyquinolin-3-yl)propanoic acid (HQA) into two different membrane proteins by heterologous expression in E. coli. Fluorescence and NMR experiments demonstrate complete replacement of the natural amino acid with HQA and stable metal chelation by the mutated proteins. Evidence of site-specific intra- and inter-molecular PREs by NMR in micelle solutions sets the stage for the use of HQA incorporation in solid-state NMR structure determinations of membrane proteins in phospholipid bilayers.

  17. Paramagnetic relaxation enhancement of membrane proteins by incorporation of the metal-chelating unnatural amino acid 2-amino-3-(8-hydroxyquinolin-3-yl)propanoic acid (HQA)

    The use of paramagnetic constraints in protein NMR is an active area of research because of the benefits of long-range distance measurements (>10 Å). One of the main issues in successful execution is the incorporation of a paramagnetic metal ion into diamagnetic proteins. The most common metal ion tags are relatively long aliphatic chains attached to the side chain of a selected cysteine residue with a chelating group at the end where it can undergo substantial internal motions, decreasing the accuracy of the method. An attractive alternative approach is to incorporate an unnatural amino acid that binds metal ions at a specific site on the protein using the methods of molecular biology. Here we describe the successful incorporation of the unnatural amino acid 2-amino-3-(8-hydroxyquinolin-3-yl)propanoic acid (HQA) into two different membrane proteins by heterologous expression in E. coli. Fluorescence and NMR experiments demonstrate complete replacement of the natural amino acid with HQA and stable metal chelation by the mutated proteins. Evidence of site-specific intra- and inter-molecular PREs by NMR in micelle solutions sets the stage for the use of HQA incorporation in solid-state NMR structure determinations of membrane proteins in phospholipid bilayers

  18. Compared with Daily, Weekly n–3 PUFA Intake Affects the Incorporation of Eicosapentaenoic Acid and Docosahexaenoic Acid into Platelets and Mononuclear Cells in Humans 1 2 3

    Browning, Lucy M; Walker, Celia G.; Mander, Adrian P; West, Annette L.; Gambell, Joanna; Madden, Jackie; Calder, Philip C.; Jebb, Susan A.

    2014-01-01

    Consumption of oily fish is sporadic, whereas controlled intervention studies of n–3 (ω-3) fatty acids usually provide capsules containing eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) as a daily dose. This methodologic study explored whether there are differences in the short-, medium-, and long-term incorporation of EPA and DHA into blood plasma and cells with the provision of identical amounts of EPA and DHA, equivalent to 2 oily fish servings per week (or 6.54 g/wk EPA and DH...

  19. Effect of Hyaluronic Acid Incorporation Method on the Stability and Biological Properties of Polyurethane-Hyaluronic Acid Biomaterials

    Ruiz, Amaliris; Rathnam, Kashmila R.; Masters, Kristyn S.

    2013-01-01

    The high failure rate of small diameter vascular grafts continues to drive the development of new materials and modification strategies that address this clinical problem, with biomolecule incorporation typically achieved via surface-based modification of various biomaterials. In this work, we examined whether the method of biomolecule incorporation (i.e., bulk vs. surface modification) into a polyurethane (PU) polymer impacted biomaterial performance in the context of vascular applications. ...

  20. Incorporation of radioactive pyrimidic precursors into the deoxyribonucleic acids of Chlorella pyrenoidosa

    (2-14C) thymidine is incorporated into the nuclear and chloroplastic DNAs of Chlorella pyrenoidosa, strain Emerson. (me-14C) thymidine is incorporated into chloroplastic DNA, but never into nuclear DNA. The radioactivity is still found to be associated with thymine. When the cultures are supplied with (2-14C) uracil, both DNAs are radioactive and the radioactivity is found to be associated with thymine, cytosine, and, for the nuclear DNA, 5-methylcytosine. (2-14C) deoxycytidine is also incorporated into both DNAs, but their specific activity is very low. These results suggest that thymidine is specifically incorporated into chloroplastic DNA of the strain Emerson. In the case of the nuclear DNA, they suggest the existence of a thymidine demethylation mechanism, probably localized in the cytoplasm. (orig.)

  1. Chitosan-clodronate nanoparticles loaded in poloxamer gel for intra-articular administration.

    Russo, E; Gaglianone, N; Baldassari, S; Parodi, B; Croce, I; Bassi, A M; Vernazza, S; Caviglioli, G

    2016-07-01

    This work was based on the study of an intra-articular delivery system constituted by a poloxamer gel vehiculating clodronate in chitosan nanoparticles. This system has been conceived to obtain a specific and controlled release of clodronate in the joints to reduce the arthritis rheumatoid degenerative effect. Clodronate (CLO) is a first-generation bisphosphonate with anti-inflammatory properties, inhibiting the cytokine and NO secretion from macrophages, therefore causing apoptosis in these cells. This is related to its ability to be metabolized by cells and converted into a cytotoxic intermediate as a non-hydrolysable analogue of ATP. Chitosan (CHI) was used to develop nanosystems, by ionotropic gelation induced by clodronate itself. A fractional factorial experimental design allowed us to obtain nanoparticles, the diameter of which ranged from 200 to 300nm. Glutaraldehyde was used to increase nanoparticle stability and modify the drug release profile. The zeta potential value of crosslinked nanopaparticles was 21.0mV±1.3, while drug loading was 31.0%±5.4 w/w; nanoparticle yield was 18.2%±1.8 w/w, the encapsulation efficiency was 48.8%±9.9 w/w. Nanoparticles were homogenously loaded in a poloxamer sol, and the drug delivery system is produced in-situ after local administration, when sol become gel at physiological temperature. The properties of poloxamer gels containing CHI-CLO nanoparticles, such as viscosity, gelation temperature and drug release properties, were evaluated. In vitro studies were conducted to evaluate the effects of these nanoparticles on a human monocytic cell line (THP1). The results showed that this drug delivery system is more efficient, with respect to the free drug, to counteract the inflammatory process characteristic of several degenerative diseases. PMID:26998870

  2. Quantification of aldehyde impurities in poloxamer by 1H NMR spectrometry

    This work presents a fast and simple quantitative method for impurity determination of acetaldehyde and propionaldehyde in poloxamer 188 by proton nuclear magnetic resonance spectroscopy (1H NMR). The sample is dissolved in D2O with DCl and analyzed with a 600 MHz NMR spectrometer. Data processing, including filtering by convolution of spectra with a triangular function and integration, is performed in MATLAB. The repeated studies of one sample, including automatic gradient shimming and data processing, revealed a relative standard deviation (R.S.D.) of 2.8%. For the reproducibility, also including sample preparation, the R.S.D. was less than 10%. The predictability of a linear calibration model was estimated by the root mean square error of prediction from leave-one-out cross-validation (RMSECV). Using 64 scans, RMSECV was found to be 7.2 and 5.5 μg g-1 for acetaldehyde and propionaldehyde respectively for a 4.3-min acquisition time. The limits of detection, defined as three times the noise, reached 19 and 15 μg g-1 respectively under the same experimental conditions. These limits are sufficient to quantify 80 and 100 μg g-1 of the impurities, which has been found to be the maximum allowed content in the poloxamer for some medical applications. Thus the method has the potential to replace the current liquid chromatography (LC) method for impurity determination of acetaldehyde and propionaldehyde in poloxamer, which is time-consuming and includes a work-up procedure involving many steps

  3. Development of an Unnatural Amino Acid Incorporation System in the Actinobacterial Natural Product Producer Streptomyces venezuelae ATCC 15439.

    He, Jingxuan; Van Treeck, Briana; Nguyen, Han B; Melançon, Charles E

    2016-02-19

    Many Actinobacteria, most notably Streptomyces, produce structurally diverse bioactive natural products, including ribosomally synthesized peptides, by multistep enzymatic pathways. The use of site-specific genetic incorporation of unnatural amino acids to investigate and manipulate the functions of natural product biosynthetic enzymes, enzyme complexes, and ribosomally derived peptides in these organisms would have important implications for drug discovery and development efforts. Here, we have designed, constructed, and optimized unnatural amino acid systems capable of incorporating p-iodo-l-phenylalanine and p-azido-l-phenylalanine site-specifically into proteins in the model natural product producer Streptomyces venezuelae ATCC 15439. We observed notable differences in the fidelity and efficiency of these systems between S. venezuelae and previously used hosts. Our findings serve as a foundation for using an expanded genetic code in Streptomyces to address questions related to natural product biosynthesis and mechanism of action that are relevant to drug discovery and development. PMID:26562751

  4. Imaging plasma docosahexaenoic acid (dha incorporation into the brain in vivo, as a biomarker of brain DHA: Metabolism and neurotransmission

    Rapoport Stanley I.

    2011-09-01

    Full Text Available Docosahexaenoic acid (DHA is critical for normal brain structure and function, and its brain concentration depends on dietary DHA content and hepatic conversion from its dietary derived n-3 precursor, a-linolenic acid (α-LNA. We developed an in vivo method in rats using quantitative autoradiography to image incorporation into brain of unesterified plasma DHA, and showed that the incorporation rate equals the rate of brain metabolic DHA consumption. Thus, quantitative imaging of DHA incorporation from plasma into brain can be used as a biomarker of brain DHA metabolism and neurotransmission. The method has been extended to humans with the use of positron emission tomography (PET. Furthermore, imaging in unanesthetized rats using DHA incorporation as a biomarker in response to N-methyl-D-aspartate (NMDA administration confirms that regional DHA signaling is independent of extracellular calcium, and likely mediated by a calcium-independent phospholipase A2 (iPLA2. Studies in mice in which iPLA2-VIA (β was knocked out confirmed that this enzyme is critical for baseline and muscarinic cholinergic signaling involving DHA.

  5. Residue-specific Incorporation of Noncanonical Amino Acids into Model Proteins Using an Escherichia coli Cell-free Transcription-translation System.

    Worst, Emanuel G; Exner, Matthias P; De Simone, Alessandro; Schenkelberger, Marc; Noireaux, Vincent; Budisa, Nediljko; Ott, Albrecht

    2016-01-01

    The canonical set of amino acids leads to an exceptionally wide range of protein functionality. Nevertheless, the set of residues still imposes limitations on potential protein applications. The incorporation of noncanonical amino acids can enlarge this scope. There are two complementary approaches for the incorporation of noncanonical amino acids. For site-specific incorporation, in addition to the endogenous canonical translational machineries, an orthogonal aminoacyl-tRNA-synthetase-tRNA pair must be provided that does not interact with the canonical ones. Consequently, a codon that is not assigned to a canonical amino acid, usually a stop codon, is also required. This genetic code expansion enables the incorporation of a noncanonical amino acid at a single, given site within the protein. The here presented work describes residue-specific incorporation where the genetic code is reassigned within the endogenous translational system. The translation machinery accepts the noncanonical amino acid as a surrogate to incorporate it at canonically prescribed locations, i.e., all occurrences of a canonical amino acid in the protein are replaced by the noncanonical one. The incorporation of noncanonical amino acids can change the protein structure, causing considerably modified physical and chemical properties. Noncanonical amino acid analogs often act as cell growth inhibitors for expression hosts since they modify endogenous proteins, limiting in vivo protein production. In vivo incorporation of toxic noncanonical amino acids into proteins remains particularly challenging. Here, a cell-free approach for a complete replacement of L-arginine by the noncanonical amino acid L-canavanine is presented. It circumvents the inherent difficulties of in vivo expression. Additionally, a protocol to prepare target proteins for mass spectral analysis is included. It is shown that L-lysine can be replaced by L-hydroxy-lysine, albeit with lower efficiency. In principle, any

  6. Single cell protein production by penicillium expansum incorporating of acid hydrolysate of rice husk in medium

    The aim of the research work is to bioconversion of rice husk to single cell protein by penicillium expansum. The rice husk was degraded chemically using sulphuric acid and perchloric acid with various concentrations (0.15, 0.30, 0.45, and 0.60 N) to fermentable sugars and these were used as substrate for the production of single cell protein by penicillium expansum. It was observed that the amount of single cell protein is higher in case of perchloric acid hydrolysate in comparison to sulphuric acid hydrolysate, while the protein content of single cell protein is higher in sulphuric acid hydrolysate. The single cell protein of penicillium expansum contains nearly all essential amino acids while it free from aflatoxin. (author)

  7. Incorporation of N-amidino-pyroglutamic acid into peptides using intramolecular cyclization of alpha-guanidinoglutaric acid.

    Burov, Sergey; Moskalenko, Yulia; Dorosh, Marina; Shkarubskaya, Zoya; Panarin, Evgeny

    2009-11-01

    N-terminal modification of peptides by unnatural amino acids significantly affects their enzymatic stability, conformational properties and biological activity. Application of N-amidino-amino acids, positively charged under physiological conditions, can change peptide conformation and its affinity to the corresponding receptor. In this article, we describe synthesis of short peptides, containing a new building block-N-amidino-pyroglutamic acid. Although direct guanidinylation of pyroglutamic acid and oxidation of N-amidino-proline using RuO(4) did not produce positive results, N-amidino-Glp-Phe-OH was synthesized on Wang polymer by cyclization of alpha-guanidinoglutaric acid residue. In the course of synthesis, it was found that literature procedure of selective Boc deprotection using TMSOTf/TEA reagent is accompanied by concomitant side reaction of triethylamine alkylation by polymer linker fragment. It should be mentioned that independently from cyclization time and coupling agent (DIC or HCTU), the lactam formation was incomplete. Separation of the cyclic product from the linear precursor was achieved by HPLC in ammonium formate buffer at pH 6. HPLC analysis showed N-amidino-Glp-Phe-OH stability at acidic and physiological pH and fast ring opening in water solution at pH 9. The suggested method of N-amidino-Glp residue formation can be applied in the case of short peptide chains, whereas synthesis of longer ones will require fragment condensation approach. PMID:19739127

  8. 苯扎贝特对poloxamer 407诱导的昆明小鼠血脂的影响%Hypolipidemic effects of Bezafibrate on poloxamer 407-induced hyperlipidemic Kunming mouse model

    李正阳; 董瓅瑾; 赵艳威; 池广鹏; 宋光明

    2012-01-01

    [目的]研究苯扎贝特对poloxamer407(P407)诱导的新型高血脂小鼠的血脂影响.[方法]昆明小鼠给予苯扎贝特(50或100 mg/kg)灌胃,连续3d,末次给药1h后,给予动物腹腔注射P407,0.3 g/kg,于注射后的4、24及48h,取血测定甘油三酯和胆固醇含量,注射P407后24h的血样同时测定高密度脂蛋白-胆固醇的含量.[结果]在造模前给予苯扎贝特处理,能明显降低高血脂动物的血清甘油三酯和胆固醇水平,且剂量关系明显,此作用可维持至造模后的48h.高血脂造模后的24h,苯扎贝特能明显升高模型动物的血清高密度脂蛋白-胆固醇水平.[结论]苯扎贝特对于P407诱导的高血脂昆明小鼠模型具有降脂作用,此模型可用于贝特类药物的研究.%[Objective]To study the effect of Bezafibrate on poloxamer 407-induced new hyperlipidemic mouse model. [Methods] Before a bolus of poloxamer 407 (0.3 g/kg) was intraperitoneally dosed, Kunming mice were pretreated orally with Bezafibrate (50 or 100mg/kg) for 3 days. Blood triacylglycerol (TG) and cholesterol (CHO) were measured at 0, 4, 24 and 48 h after the poloxamer 407 injection, and the blood HDL-CHO level of Time24h, were measured. [Results] Poloxamer407 injection caused hyperlipidemia in Kunming mice. Bezafibrate decreased dose-dependently blood TG and CHO, and this hypoipidemic effect lasted up to 48 h after injection. Moreover, Bezafibrate increased blood HDL-CHO in poloxamer 407 induced hyperlipidemic mice. [Conclusions] Bezafibrate could have a hypolipidemic effects in poloxamer 407induced hyperlipidemic Kuonming mice. This animal model may be used to evaluate the lipid-lowering effects of statin drugs.

  9. Characterization of Active Packaging Films Made from Poly(Lactic Acid/Poly(Trimethylene Carbonate Incorporated with Oregano Essential Oil

    Dong Liu

    2016-05-01

    Full Text Available Antimicromial and antioxidant bioactive films based on poly(lactic acid/poly(trimenthylene carbonate films incorporated with different concentrations of oregano essential oil (OEO were prepared by solvent casting. The antimicrobial, antioxidant, physical, thermal, microstructural, and mechanical properties of the resulting films were examined. Scanning electron microscopy analysis revealed that the cross-section of films became rougher when OEO was incorporated into PLA/PTMC blends. Differential scanning calorimetry analysis indicated that crystallinity of PLA phase decreased by the addition of OEO, but this did not affect the thermal stability of the films. Water vapor permeability of films slightly increased with increasing concentration of OEO. However, active PLA/PTMC/OEO composite films showed adequate barrier properties for food packaging application. The antimicrobial and antioxidant capacities were significantly improved with the incorporation of OEO (p < 0.05. The results demonstrated that an optimal balance between the mechanical, barrier, thermal, antioxidant, and antimicrobial properties of the films was achieved by the incorporation of 9 wt % OEO into PLA/PTMC blends.

  10. Characterization of Active Packaging Films Made from Poly(Lactic Acid)/Poly(Trimethylene Carbonate) Incorporated with Oregano Essential Oil.

    Liu, Dong; Li, Hongli; Jiang, Lin; Chuan, Yongming; Yuan, Minglong; Chen, Haiyun

    2016-01-01

    Antimicromial and antioxidant bioactive films based on poly(lactic acid)/poly(trimenthylene carbonate) films incorporated with different concentrations of oregano essential oil (OEO) were prepared by solvent casting. The antimicrobial, antioxidant, physical, thermal, microstructural, and mechanical properties of the resulting films were examined. Scanning electron microscopy analysis revealed that the cross-section of films became rougher when OEO was incorporated into PLA/PTMC blends. Differential scanning calorimetry analysis indicated that crystallinity of PLA phase decreased by the addition of OEO, but this did not affect the thermal stability of the films. Water vapor permeability of films slightly increased with increasing concentration of OEO. However, active PLA/PTMC/OEO composite films showed adequate barrier properties for food packaging application. The antimicrobial and antioxidant capacities were significantly improved with the incorporation of OEO (p < 0.05). The results demonstrated that an optimal balance between the mechanical, barrier, thermal, antioxidant, and antimicrobial properties of the films was achieved by the incorporation of 9 wt % OEO into PLA/PTMC blends. PMID:27240336

  11. MORTAR INCORPORATING SUPPLEMENTARY CEMENTITIOUS MATERIALS: STRENGTH, ISOTHERMAL CALORIMETRY AND ACIDS ATTACK

    Y. Senhadji

    2016-05-01

    Full Text Available Supplementary cementitious materials (SCMs prove to be effective to meet most of the requirements of durable concrete and leads to a significant reduction in CO2 emissions. This research studies the effect different SCMs (natural pozzolan (PN/ limestone fine (FC at various remplacement levels on the physical and mechano-chemical resistance of blended mortar. The paper primarily deals with the characteristics of these materials, including heat of hydration, strength and effects of aggressive chemical environments (using sulphuric acid and nitric acid. Over 6 mixes were made and compared to the control mix. Tests were conducted at different ages up to 360 days. The experimental results in general showed that Algerian mineral admixtures (PN/FC were less vulnerable to nitric and sulphuric acid attack and improved the properties of mortars, but at different rates depending on the quantity of binder.

  12. Fatty acids from membrane lipids become incorporated into lipid bodies during Myxococcus xanthus differentiation.

    Swapna Bhat

    Full Text Available Myxococcus xanthus responds to amino acid limitation by producing fruiting bodies containing dormant spores. During development, cells produce triacylglycerides in lipid bodies that become consumed during spore maturation. As the cells are starved to induce development, the production of triglycerides represents a counterintuitive metabolic switch. In this paper, lipid bodies were quantified in wild-type strain DK1622 and 33 developmental mutants at the cellular level by measuring the cross sectional area of the cell stained with the lipophilic dye Nile red. We provide five lines of evidence that triacylglycerides are derived from membrane phospholipids as cells shorten in length and then differentiate into myxospores. First, in wild type cells, lipid bodies appear early in development and their size increases concurrent with an 87% decline in membrane surface area. Second, developmental mutants blocked at different stages of shortening and differentiation accumulated lipid bodies proportionate with their cell length with a Pearson's correlation coefficient of 0.76. Third, peripheral rods, developing cells that do not produce lipid bodies, fail to shorten. Fourth, genes for fatty acid synthesis are down-regulated while genes for fatty acid degradation are up regulated. Finally, direct movement of fatty acids from membrane lipids in growing cells to lipid bodies in developing cells was observed by pulse labeling cells with palmitate. Recycling of lipids released by Programmed Cell Death appears not to be necessary for lipid body production as a fadL mutant was defective in fatty acid uptake but proficient in lipid body production. The lipid body regulon involves many developmental genes that are not specifically involved in fatty acid synthesis or degradation. MazF RNA interferase and its target, enhancer-binding protein Nla6, appear to negatively regulate cell shortening and TAG accumulation whereas most cell-cell signals activate these

  13. Incorporation and effects of punicic acid on muscle and adipose tissues of rats

    Illana Louise Pereira de MELO; de Oliveira e Silva, Ana Mara; Eliane Bonifácio Teixeira de CARVALHO; Luciana Tedesco YOSHIME; Sattler, José Augusto Gasparotto; Jorge MANCINI-FILHO

    2016-01-01

    Background This study evaluated the effect of pomegranate seed oil (PSO) supplementation, rich in punicic acid (55 %/C18:3-9c,11 t,13c/CLNA), on the lipid profile and on the biochemical and oxidative parameters in the gastrocnemius muscle and adipose tissues of healthy rats. Linseed oil (LO), rich in linolenic acid (52 %/C18:3-9c12c15c/LNA) was used for comparison. Methods Male Wistar rats (n = 56) were distributed in seven groups: control (water); LNA 1 %, 2 % and 4 % (treated with LO); CLNA...

  14. Preparation and characterization of electrospun PLCL/Poloxamer nanofibers and dextran/gelatin hydrogels for skin tissue engineering.

    Jian-feng Pan

    Full Text Available In this study, two different biomaterials were fabricated and their potential use as a bilayer scaffold for skin tissue engineering applications was assessed. The upper layer biomaterial was a Poly(ε-caprolactone-co-lactide/Poloxamer (PLCL/Poloxamer nanofiber membrane fabricated using electrospinning technology. The PLCL/Poloxamer nanofibers (PLCL/Poloxamer, 9/1 exhibited strong mechanical properties (stress/strain values of 9.37 ± 0.38 MPa/187.43 ± 10.66% and good biocompatibility to support adipose-derived stem cells proliferation. The lower layer biomaterial was a hydrogel composed of 10% dextran and 20% gelatin without the addition of a chemical crosslinking agent. The 5/5 dextran/gelatin hydrogel displayed high swelling property, good compressive strength, capacity to present more than 3 weeks and was able to support cells proliferation. A bilayer scaffold was fabricated using these two materials by underlaying the nanofibers and casting hydrogel to mimic the structure and biological function of native skin tissue. The upper layer membrane provided mechanical support in the scaffold and the lower layer hydrogel provided adequate space to allow cells to proliferate and generate extracellular matrix. The biocompatibility of bilayer scaffold was preliminarily investigated to assess the potential cytotoxicity. The results show that cell viability had not been affected when cocultured with bilayer scaffold. As a consequence, the bilayer scaffold composed of PLCL/Poloxamer nanofibers and dextran/gelatin hydrogels is biocompatible and possesses its potentially high application prospect in the field of skin tissue engineering.

  15. Evaluation of a novel thermosensitive heparin-poloxamer hydrogel for improving vascular anastomosis quality and safety in a rabbit model.

    Ying-Zheng Zhao

    Full Text Available Despite progress in the design of advanced surgical techniques, stenosis recurs in a large percentage of vascular anastomosis. In this study, a novel heparin-poloxamer (HP hydrogel was designed and its effects for improving the quality and safety of vascular anastomosis were studied. HP copolymer was synthesized and its structure was confirmed by Fourier transform infrared spectroscopy (FTIR and nuclear magnetic resonance spectroscopy ((1H-NMR. Hydrogels containing HP were prepared and their important characteristics related to the application in vascular anastomosis including gelation temperature, rheological behaviour and micromorphology were measured. Vascular anastomosis were performed on the right common carotid arteries of rabbits, and the in vivo efficiency and safety of HP hydrogel to achieve vascular anastomosis was verified and compared with Poloxamer 407 hydrogel and the conventional hand-sewn method using Doppler ultrasound, CT angiograms, scanning electron microscopy (SEM and histological technique. Our results showed that HP copolymer displayed special gel-sol-gel phase transition behavior with increasing temperature from 5 to 60 °C. HP hydrogel prepared from 18 wt% HP solution had a porous sponge-like structure, with gelation temperature at approximately 38 °C and maximum elastic modulus at 10,000 Pa. In animal studies, imaging and histological examination of rabbit common jugular artery confirmed that HP hydrogel group had similar equivalent patency, flow and burst strength as Poloxamer 407 group. Moreover, HP hydrogel was superior to poloxamer 407 hydrogel and hand-sewn method for restoring the functions and epithelial structure of the broken vessel junctions after operation. By combining the advantages of heparin and poloxamer 407, HP hydrogel holds high promise for improving vascular anastomosis quality and safety.

  16. Preparation of Monolithic Ti-incorporated Mesoporous SilicaMaterials via Tartaric Acid Templated Sol-gel Process

    PANG Jie-Bina; QIU Kun-Yuan; WEI Yen

    2001-01-01

    Monolithie and transparent Ti-incorporated mesoporoussilica materials of largesize(e.g.2mm) in dimension have been prepared with tartaric acid(TA)as template via sol-gel reac-tions of tetraethyl orthosilicate (TEOS)and tetrabutyl titanat (TBT).The materials are characterized by infrared (IR),nitrogen adsorption-desorption istherms,powder X-ray diffraction(XRD)and transmission electron microscopy (TEM).The results indicate that the monolithic materials ex-hibit large specific surface(ca.1200m 2/g)and pore volumes (ca.0.900cm3/g).

  17. Antimicrobial activity of nisin incorporated in pectin and polylactic acid composite films against Listeria monocytogenes

    Extruded composite films from 20% pectin and 80% polylactic acids (PLA) were developed and nisin was loaded into films by a diffusion post extrusion. Inhibitory activities of the films against Listeria monocytogenes were evaluated in brain heart infusion (BHI) broth, liquid egg white and orange juic...

  18. Incorporated sarcolemmal fish oil fatty acids shorten pig ventricular action potentials

    Verkerk, A.O.; Ginneken, van A.C.G.; Berecki, G.; Ruijter, den H.M.; Schumacher, C.A.; Veldkamp, M.W.; Baartscheer, A.; Casini, S.; Opthof, T.; Hovenier, R.; Fiolet, J.W.T.; Zock, P.L.; Coronel, R.

    2006-01-01

    Background: Omega-3 polyunsaturated fatty acids (W-PUFAs) from fish oil reduce the risk of sudden death presumably by preventing life-threatening arrhythmias. Acutely administered omega 3-PUFAs modulate the activity of several cardiac ion channels, but the chronic effects of a diet enriched with fis

  19. Lactobacillus plantarum as source of conjugated linoleic acid: Effect of pH, incubation Temperature and inulin incorporation.

    Carmen Soto

    2013-07-01

    Full Text Available The effect of pH and temperature, and inulin use, on the growth andthe fatty acid profile of Lactobacillus plantarum strain wereevaluated. The best results were obtained at 6.5 pH broth, producing3.2 g/L of biomass and about 20% of conjugated linoleic acid(CLA in the cell lipids. Similar growth was observed with 37 and45ºC, but a low CLA content (10.6% was achieved at 45°C. In thecase of inulin incorporation, a low biomass concentration (1 g/Land low production of CLA (12.4% were observed. These resultssuggest a pH and temperature dependence on CLA production bythe microorganism.

  20. Synthetic poly(amino acid) hydrogels with incorporated cell-adhesion peptides for tissue engineering

    Studenovská, Hana; Vodička, Petr; Proks, Vladimír; Hlučilová, Jana; Motlík, Jan; Rypáček, František

    2010-01-01

    Roč. 4, č. 6 (2010), s. 454-463. ISSN 1932-6254 R&D Projects: GA AV ČR KJB400500801; GA MŠk 1M0538 Institutional research plan: CEZ:AV0Z40500505; CEZ:AV0Z50450515 Keywords : polyamino acid * hydrogel * porosity Subject RIV: CD - Macromolecular Chemistry Impact factor: 3.534, year: 2010

  1. High affinity nucleic acid aptamers for streptavidin incorporated into bi-specific capture ligands

    Tahiri-Alaoui, Abdessamad; Frigotto, Laura; Manville, Nick; Ibrahim, Jamal; Romby, Pascale; James, William

    2002-01-01

    We have isolated 2′-Fluoro-substituted RNA aptamers that bind to streptavidin (SA) with an affinity around 7 ± 1.8 nM, comparable with that of recently described peptide aptamers. Binding to SA was not prevented by prior saturation with biotin, enabling nucleic acid aptamers to form useful ternary complexes. Mutagenesis, secondary structure analysis, ribonuclease footprinting and deletion analysis provided evidence for the essential structural features of SA-binding aptamers. In order to prov...

  2. Lipoxygenase- and cyclooxygenase-reaction products and incorporation into glycerolipids or radiolabeled arachidonic acid in the bovine retina

    The metabolism of radiolabeled arachidonic acid (AA) by the intact bovine retina in vitro has been studied. Synthesis of prostaglandins (PGs) and hydroxyeicosatetraenoic acids (HETEs), and incorporation of AA into glycerolipids has been measured by reverse-phase and straight-phase high performance liquid chromatography with flow scintillation detection, and by thin-layer chromatography. AA was actively acylated into glycerolipids, particularly triglycerides, phosphatidylcholine and phosphatidylinositol. AA was also converted to the major PGs, PGF2 alpha, PGE2, PGD2, 6-keto-PGF1 alpha and TXB2, and to the lipoxygenase reaction products, 12-HETE, 5-HETE, and other monohydroxy isomers. Approximately 6% of the radiolabeled AA was converted to eicosanoids. The synthesis of HETEs was inhibited in a concentration-dependent manner (IC50 . 8.3 nM) by nordihydroguaiaretic acid (NDGA). PG synthesis was inhibited by aspirin (10 microM), indomethacin (1 microM) and NDGA (IC50 . 380 nM). Metabolism of AA via lipoxygenase, cyclooxygenase and activation-acylation was inhibited by boiling retinal tissue prior to incubation. These studies demonstrate an active system for the uptake and utilization of AA in the bovine retina, and provide the first evidence of lipoxygenase-mediated metabolism of AA, resulting in the synthesis of mono-hydroxyeicosatetraenoic acids, in the retina

  3. Evidence for a universal pathway of abscisic acid biosynthesis in higher plants from 18O incorporation patterns

    Previous labeling studies of abscisic acid (ABA) with 18O2 have been mainly conducted with water-stressed leaves. In this study, 18O incorporation into ABA of stressed leaves of various species was compared with 18O labeling of ABA of turgid leaves and of fruit tissue in different stages of ripening. In stressed leaves of all six species investigated, avocado (Persea americana), barley (Hordeum vulgare), bean (Phaseolus vulgaris), cocklebur (Xanthium strumarium), spinach (Spinacia oleracea), and tobacco (Nicotiana tabacum), 18O was most abundant in the carboxyl group, whereas incorporation of a second and third 18O in the oxygen atoms on the ring of ABA was much less prominent after 24 h in 18O2. ABA from turgid bean leaves showed significant 18O incorporation, again with highest 18O enrichment in the carboxyl group. On the basis of 18O-labeling patterns observed in ABA from different tissues it is concluded that, despite variations in precusor pool sizes and intermediate turnover rates, there is a universal pathway of ABA biosynthesis in higher plants which involves cleavage of a larger precursor molecule, presumably an oxygenated carotenoid

  4. Evidence for a universal pathway of abscisic acid biosynthesis in higher plants from sup 18 O incorporation patterns

    Zeevaart, J.A.D.; Heath, T.G.; Gage, D.A. (Michigan State University, East Lansing (USA))

    1989-12-01

    Previous labeling studies of abscisic acid (ABA) with {sup 18}O{sub 2} have been mainly conducted with water-stressed leaves. In this study, {sup 18}O incorporation into ABA of stressed leaves of various species was compared with {sup 18}O labeling of ABA of turgid leaves and of fruit tissue in different stages of ripening. In stressed leaves of all six species investigated, avocado (Persea americana), barley (Hordeum vulgare), bean (Phaseolus vulgaris), cocklebur (Xanthium strumarium), spinach (Spinacia oleracea), and tobacco (Nicotiana tabacum), {sup 18}O was most abundant in the carboxyl group, whereas incorporation of a second and third {sup 18}O in the oxygen atoms on the ring of ABA was much less prominent after 24 h in {sup 18}O{sub 2}. ABA from turgid bean leaves showed significant {sup 18}O incorporation, again with highest {sup 18}O enrichment in the carboxyl group. On the basis of {sup 18}O-labeling patterns observed in ABA from different tissues it is concluded that, despite variations in precusor pool sizes and intermediate turnover rates, there is a universal pathway of ABA biosynthesis in higher plants which involves cleavage of a larger precursor molecule, presumably an oxygenated carotenoid.

  5. Hair dye-incorporated poly-γ-glutamic acid/glycol chitosan nanoparticles based on ion-complex formation

    Lee HY

    2011-11-01

    Full Text Available Hye-Young Lee1,*, Young-IL Jeong2,*, Ki-Choon Choi31Anyang Science University, Anyang, Gyeonggi, South Korea; 2Chonnam National University Hwasun Hospital, Jeonnam, South Korea; 3Grassland and Forages Research Center, National Institute of Animal Science, Rural Development Administration, Chungnam, South Korea*These authors contributed equally to this work.Background: p-Phenylenediamine (PDA or its related chemicals are used more extensively than oxidative hair dyes. However, permanent hair dyes such as PDA are known to have potent contact allergy reactions in humans, and severe allergic reactions are problematic.Methods: PDA-incorporated nanoparticles were prepared based on ion-complex formation between the cationic groups of PDA and the anionic groups of poly(γ-glutamic acid (PGA. To reinforce PDA/PGA ion complexes, glycol chitosan (GC was added. PDA-incorporated nanoparticles were characterized using field-emission scanning electron microscopy, Fourier-transform infrared (FT-IR spectroscopy, dynamic light scattering, and powder X-ray diffractometry (XRD.Results: Nanoparticles were formed by ion-complex formation between the amine groups of PDA and the carboxyl groups of PGA. PDA-incorporated nanoparticles are small in size (<100 nm, and morphological observations showed spherical shapes. FT-IR spectra results showed that the carboxylic acid peak of PGA decreased with increasing PDA content, indicating that the ion complexes were formed between the carboxyl groups of PGA and the amine groups of PDA. Furthermore, the intrinsic peak of the carboxyl groups of PGA was also decreased by the addition of GC. Intrinsic crystalline peaks of PDA were observed by XRD. This crystalline peak of PDA was completely nonexistent when nanoparticles were formed by ion complex between PDA, PGA, and GC, indicating that PDA was complexed with PGA and no free drug existed in the formulation. During the drug-release experiment, an initial burst release of PDA was

  6. Synthetic Cyclolipopeptides Selective against Microbial, Plant and Animal Cell Targets by Incorporation of D-Amino Acids or Histidine

    Vilà, Sílvia; Badosa, Esther; Montesinos, Emilio; Planas, Marta; Feliu, Lidia

    2016-01-01

    Cyclolipopeptides derived from the antimicrobial peptide c(Lys-Lys-Leu-Lys-Lys-Phe-Lys-Lys-Leu-Gln) (BPC194) were prepared on solid-phase and screened against four plant pathogens. The incorporation at Lys5 of fatty acids of 4 to 9 carbon atoms led to active cyclolipopeptides. The influence on the antimicrobial activity of the Lys residue that is derivatized was also evaluated. In general, acylation of Lys1, Lys2 or Lys5 rendered the sequences with the highest activity. Incorporation of a D-amino acid maintained the antimicrobial activity while significantly reduced the hemolysis. Replacement of Phe with a His also yielded cyclolipopeptides with low hemolytic activity. Derivatives exhibiting low phytotoxicity in tobacco leaves were also found. Interestingly, sequences with or without significant activity against phytopathogenic bacteria and fungi, but with differential hemolysis and phytotoxicity were identified. Therefore, this study represents an approach to the development of bioactive peptides with selective activity against microbial, plant and animal cell targets. These selective cyclolipopeptides are candidates useful not only to combat plant pathogens but also to be applied in other fields. PMID:27008420

  7. Improved thrombin binding aptamer by incorporation of a single unlocked nucleic acid monomer

    Pasternak, Anna; Hernandez, Frank J; Rasmussen, Lars Melholt;

    2011-01-01

    A 15-mer DNA aptamer (named TBA) adopts a G-quadruplex structure that strongly inhibits fibrin-clot formation by binding to thrombin. We have performed thermodynamic analysis, binding affinity and biological activity studies of TBA variants modified by unlocked nucleic acid (UNA) monomers. UNA...... that a UNA monomer is allowed in many positions of the aptamer without significantly changing the thrombin-binding properties. The biological effect of a selection of the modified aptamers was tested by a thrombin time assay and showed that most of the UNA-modified TBAs possess anticoagulant properties...

  8. Enhanced anticancer potency using an acid-responsive ZnO-incorporated liposomal drug-delivery system

    Tripathy, Nirmalya; Ahmad, Rafiq; Ko, Hyun Ah; Khang, Gilson; Hahn, Yoon-Bong

    2015-02-01

    The development of stimuli-responsive nanocarriers is becoming important in chemotherapy. Liposomes, with an appropriate triggering mechanism, can efficiently deliver their encapsulated cargo in a controlled manner. We explored the use of acid-sensitive zinc oxide nanoparticles (ZNPs) as modulators of the responsive properties of liposomes. Nanocomplexes formed by the incorporation of ZNPs in liposomes (ZNP-liposomes) were designed to demonstrate the pH-responsive release of a drug (daunorubicin) without premature drug leakage and with the maintenance of the relevant therapeutic concentrations. The nanocomplexes were spherical in shape with a narrow size distribution and showed a high drug-encapsulating efficiency. Under acidic conditions, the ZNP-liposome nanocomplexes released the loaded drug more rapidly than bare liposomes. Using flow cytometry, confocal microscopy and an MTT assay, we demonstrated that these nanocomplexes were readily taken up by cancer cells, resulting in significantly enhanced cytotoxicity. On exposure to the acidic conditions inside cancer cells, the ZNPs rapidly decomposed, releasing the entrapped drug molecules from the ZNP-liposome nanocomplexes, producing widespread cytotoxic effects. The incorporated ZNPs were multimodal in that they not only resulted in a pH-responsive drug-delivery system, but they also had a synergistic chemo-photodynamic anticancer action. This design provides a significant step towards the development of multimodal liposome structures.The development of stimuli-responsive nanocarriers is becoming important in chemotherapy. Liposomes, with an appropriate triggering mechanism, can efficiently deliver their encapsulated cargo in a controlled manner. We explored the use of acid-sensitive zinc oxide nanoparticles (ZNPs) as modulators of the responsive properties of liposomes. Nanocomplexes formed by the incorporation of ZNPs in liposomes (ZNP-liposomes) were designed to demonstrate the pH-responsive release of a drug

  9. Incorporation of small BN domains in graphene during CVD using methane, boric acid and nitrogen gas.

    Bepete, George; Voiry, Damien; Chhowalla, Manish; Chiguvare, Zivayi; Coville, Neil J

    2013-07-21

    Chemical doping of graphene with small boron nitride (BN) domains has been shown to be an effective way of permanently modulating the electronic properties in graphene. Herein we show a facile method of growing large area graphene doped with small BN domains on copper foils using a single step CVD route with methane, boric acid powder and nitrogen gas as the carbon, boron and nitrogen sources respectively. This facile and safe process avoids the use of boranes and ammonia. Optical microscopy confirmed that continuous films were grown and Raman spectroscopy confirmed changes in the electronic structure of the grown BN doped graphene. Using XPS studies we find that both B and N can be substituted into the graphene structure in the form of small BN domains to give a B-N-C system. A novel structure for the BN doped graphene is proposed. PMID:23759928

  10. Incorporation of small BN domains in graphene during CVD using methane, boric acid and nitrogen gas

    Bepete, George; Voiry, Damien; Chhowalla, Manish; Chiguvare, Zivayi; Coville, Neil J.

    2013-06-01

    Chemical doping of graphene with small boron nitride (BN) domains has been shown to be an effective way of permanently modulating the electronic properties in graphene. Herein we show a facile method of growing large area graphene doped with small BN domains on copper foils using a single step CVD route with methane, boric acid powder and nitrogen gas as the carbon, boron and nitrogen sources respectively. This facile and safe process avoids the use of boranes and ammonia. Optical microscopy confirmed that continuous films were grown and Raman spectroscopy confirmed changes in the electronic structure of the grown BN doped graphene. Using XPS studies we find that both B and N can be substituted into the graphene structure in the form of small BN domains to give a B-N-C system. A novel structure for the BN doped graphene is proposed.

  11. Extraction and transport of uranium (VI) by polymer inclusion membranes incorporating Di-(2-ethylhexyl) phosphoric acid

    Di-(2-ethylhexyl) phosphoric acid (D2EHPA) is a commonly used extraction reagent for the separation of uranium from aqueous solutions. Previous work showed the suitability of a polymer inclusion membrane (PIM) based on D2EHPA and poly(vinyl chloride) (PVC) for the extraction and transport of uranium (VI) from sulfate solutions. This study investigates the influence of the main system parameters (i.e. stirring rate, percentage of D2EHPA in the membrane, initial U(VI) concentration, membrane thickness, and concentration of sulfuric acid) on the extraction process. A comprehensive mathematical model, describing the extraction of U(VI), was developed, numerically solved and fitted to the experimental extraction data to determine the values of the extraction constant and the diffusion coefficient of the U(VI)-D2EHPA complex in the membrane. The transport properties of the PVC/D2EHPA PIM have been further improved by adding o-nitrophenyloctyl ether as plasticizer. Fluxes of U(VI) as high as 1.5 x 10-6 mol m-2 S-1 were recorded using a membrane composed of 35% D2EHPA, 10% o-nitrophenyloctyl ether and 55% PVC (m/m) from a solution containing 100 mg L-1 U(VI) in 0.1 mol L-1 H2SO4 into a solution containing 6 mol L-1H2SO4. The effect of the counter-ion on the extraction and back extraction of uranium was also investigated and the membranes were tested for durability over repeated cycles of extraction and back-extraction. It was demonstrated that a 45% D2EHPA and 55% PVC PIM (m/m) allowed U(VI) to be completely separated thermodynamically from a range of common metal cations and kinetically from iron (III). (author)

  12. Fluorescence turn-on recognition of chiral amino acids using dye incorporated β-CD functionalized AuNPs assembly

    An assembly of dye incorporated β-cyclodextrin (βCD) functionalized AuNPs for the fluorescent probing of chiral amino acids is presented. Gold nanoparticles (AuNPs) possessing a high extinction coefficient function can be used as excellent fluorescent quenchers in AuNP–fluorophore system. Inclusion of fluorescein (FL) into β-cyclodextrin (βCD) makes energy transfer to occur through the donor and quencher nearby. This energy transfer switches off by virtue of the analyte induced release of FL from β-CD cavity, which results in the fluorescence recovery of the quenched dye. Analysis suggests that the assembly of AuNPs–βCDs–FL is effective as a turn-on fluorescent probe for the chiroselective optical discrimination between D,L-tryptophan, D,L-phenyl alanine and D,L-tyrosine. The detection limits for analyzing L-tryptophan, L-phenyl alanine and L-tyrosine were found to be 0.59, 1.2 and 1.5 μM respectively. - Highlights: • Fluorescence quenching AuNP–βCD–dye assembly via energy transfer. • Energy transfer from dye to AuNPs is a SET process. • Fluorescence turn-on detection of amino acids by the competitive binding method. • Chiroselective discrimination between enantiomeric amino acids

  13. Fluorescence turn-on recognition of chiral amino acids using dye incorporated β-CD functionalized AuNPs assembly

    Aswathy, B., E-mail: aswathybv@gmail.com; Sony, G., E-mail: emailtosony@gmail.com

    2014-10-15

    An assembly of dye incorporated β-cyclodextrin (βCD) functionalized AuNPs for the fluorescent probing of chiral amino acids is presented. Gold nanoparticles (AuNPs) possessing a high extinction coefficient function can be used as excellent fluorescent quenchers in AuNP–fluorophore system. Inclusion of fluorescein (FL) into β-cyclodextrin (βCD) makes energy transfer to occur through the donor and quencher nearby. This energy transfer switches off by virtue of the analyte induced release of FL from β-CD cavity, which results in the fluorescence recovery of the quenched dye. Analysis suggests that the assembly of AuNPs–βCDs–FL is effective as a turn-on fluorescent probe for the chiroselective optical discrimination between D,L-tryptophan, D,L-phenyl alanine and D,L-tyrosine. The detection limits for analyzing L-tryptophan, L-phenyl alanine and L-tyrosine were found to be 0.59, 1.2 and 1.5 μM respectively. - Highlights: • Fluorescence quenching AuNP–βCD–dye assembly via energy transfer. • Energy transfer from dye to AuNPs is a SET process. • Fluorescence turn-on detection of amino acids by the competitive binding method. • Chiroselective discrimination between enantiomeric amino acids.

  14. Prediction of Protein Submitochondrial Locations by Incorporating Dipeptide Composition into Chou's General Pseudo Amino Acid Composition.

    Ahmad, Khurshid; Waris, Muhammad; Hayat, Maqsood

    2016-06-01

    Mitochondrion is the key organelle of eukaryotic cell, which provides energy for cellular activities. Submitochondrial locations of proteins play crucial role in understanding different biological processes such as energy metabolism, program cell death, and ionic homeostasis. Prediction of submitochondrial locations through conventional methods are expensive and time consuming because of the large number of protein sequences generated in the last few decades. Therefore, it is intensively desired to establish an automated model for identification of submitochondrial locations of proteins. In this regard, the current study is initiated to develop a fast, reliable, and accurate computational model. Various feature extraction methods such as dipeptide composition (DPC), Split Amino Acid Composition, and Composition and Translation were utilized. In order to overcome the issue of biasness, oversampling technique SMOTE was applied to balance the datasets. Several classification learners including K-Nearest Neighbor, Probabilistic Neural Network, and support vector machine (SVM) are used. Jackknife test is applied to assess the performance of classification algorithms using two benchmark datasets. Among various classification algorithms, SVM achieved the highest success rates in conjunction with the condensed feature space of DPC, which are 95.20 % accuracy on dataset SML3-317 and 95.11 % on dataset SML3-983. The empirical results revealed that our proposed model obtained the highest results so far in the literatures. It is anticipated that our proposed model might be useful for future studies. PMID:26746980

  15. Synthesis, structure, and thermodynamics of a lanthanide coordination compound incorporating 5-nitroisophthalic acid

    Highlights: ► A microporous LnMOF have been prepared and characterized. ► The standard molar enthalpy for the compound was determined to be to be −(831.4 ± 16.0) kJ · mol−1. ► Thermodynamics and kinetics of the reaction of formation of the compound were invesitigated. - Abstract: A lanthanide coordination compound, [Sm3(5-nip)4(5-Hnip)(H2O)7·9H2O]n (5-H2nip = 5-nitroisophthalic acid), has been synthesized and characterized by elemental analysis, IR, TG-DSC, and single-crystal X-ray diffraction. Structural analysis reveals that the compound features two kinds of 1D channels with guest water molecules. TG-DSC curves show that the dehydrated product of the compound exhibits high stability up to 673 K. The enthalpy change of reaction of formation in water, ΔrHmθ(l), was determined to be (27.608 ± 0.133) kJ · mol−1 at (298.15 ± 0.01) K by microcalorimetry. Based on a designed thermochemical cycle and other auxiliary thermodynamic data, the enthalpy change of reaction of formation in solid at (298.15 ± 0.01) K and the standard molar enthalpy for the compound, ΔrHmθ(s) and ΔfHmθ, were calculated to be (96.8 ± 0.8) kJ · mol−1 and (−831.4 ± 16.0) kJ · mol−1, respectively. In addition, thermodynamics and thermokinetics of the reaction of formation of the compound were investigated in water.

  16. Incorporation of hyaluronic acid into collagen scaffolds for the control of chondrocyte-mediated contraction and chondrogenesis

    Tang Shunqing [Department of Biomedical Engineering, Jinan University, Guangzhou 510632 (China); Spector, Myron [Tissue Engineering, VA Boston Healthcare System, Boston, MA 02130 (United States)

    2007-09-15

    Hyaluronic acid (HA), a principal matrix molecule in many tissues, is present in high amounts in articular cartilage. HA contributes in unique ways to the physical behavior of the tissue, and has been shown to have beneficial effects on chondrocyte activity. The goal of this study was to incorporate graduated amounts of HA into type I collagen scaffolds for the control of chondrocyte-mediated contraction and chondrogenesis in vitro. The results demonstrated that the amount of contraction of HA/collagen scaffolds by adult canine articular chondrocytes increased with the HA content of the scaffolds. The greatest amount of chondrogenesis after two weeks was found in the scaffolds which had undergone the most contraction. HA can play a useful role in adjusting the mechanical behavior of tissue engineering scaffolds and chondrogenesis in chondrocyte-seeded scaffolds.

  17. Incorporation of hyaluronic acid into collagen scaffolds for the control of chondrocyte-mediated contraction and chondrogenesis

    Hyaluronic acid (HA), a principal matrix molecule in many tissues, is present in high amounts in articular cartilage. HA contributes in unique ways to the physical behavior of the tissue, and has been shown to have beneficial effects on chondrocyte activity. The goal of this study was to incorporate graduated amounts of HA into type I collagen scaffolds for the control of chondrocyte-mediated contraction and chondrogenesis in vitro. The results demonstrated that the amount of contraction of HA/collagen scaffolds by adult canine articular chondrocytes increased with the HA content of the scaffolds. The greatest amount of chondrogenesis after two weeks was found in the scaffolds which had undergone the most contraction. HA can play a useful role in adjusting the mechanical behavior of tissue engineering scaffolds and chondrogenesis in chondrocyte-seeded scaffolds

  18. Maximising metal ions flux across a microdialysis membrane by incorporating poly-L-aspartic acid, poly-L-histidine, 8-hydroxyquinoline and ethylenediaminetetraacetic acid in the perfusion liquid

    This paper presents a study of quiescent microdialysis sampling of Cr3+, Cu2+, Ni2+ and Pb2+ involving the incorporation of poly-L-aspartic acid, poly-L-histidine, 8-hydroxyquinoline (8-HQ) and ethylenediaminetetraacetic acid (EDTA), in the perfusion liquid as an approach to maximise metal analyte flux across the microdialysis membrane. These chelating agents were individually optimised with respect to microdialysis recovery and subsequently combined in the perfusion liquid. A combination of 20% (w/v) poly-L-histidine, 0.032% (w/v) poly-L-aspartic acid and 1 mM 8-HQ achieved microdialysis recovery up to 90%. Since 1 mM EDTA achieved recoveries greater than 80% for all metals understudy, EDTA was not combined with any of the chelating agents. Under the optimal conditions of maximum metal ion flux across the microdialysis membrane, metal ions from natural and wastewater were sampled and analysed with an electrothermal atomic absorption spectrometer equipped with a Zeeman background corrector. Results showed higher concentrations of detected metal ions after microdialysis sampling compared to direct detection without sample clean-up. Incorporation of chelating agents in the microdialysis perfusion liquid enhanced metal ions recovery in real samples and achieved enrichment factors of up to 42. The study demonstrated that combining chelating agents is a good approach towards maximising metal flux across the dialysis membrane. Given that recoveries between 80 and 90% were achieved under quiescent microdialysis sampling conditions, these findings are an important development for in vivo diagnostic sampling of metal ions

  19. Oxidation and sensing of ascorbic acid and dopamine on self-assembled gold nanoparticles incorporated within polyaniline film

    Chu, Wenya; Zhou, Qun; Li, Shuangshuang; Zhao, Wei; Li, Na; Zheng, Junwei

    2015-10-01

    Electrochemical biosensors based on conducting polymers incorporated with metallic nanoparticles can greatly enhance sensitivity and selectivity. Herein, we report a facile fabrication approach for polyaniline (PAN) incorporated with a gold nanoparticle (AuNP) composite electrode by electrodeposition of PAN on a self-assembled AuNP layer on the surface of an indium tin oxide electrode. The resulting AuNP/PAN composite electrode exhibits a remarkable synergistic effect on the electrocatalytic oxidation of ascorbic acid (AA) and dopamine (DA). It is demonstrated that the oxidation reaction of AA mainly occurs at AuNPs inside the PAN film as the ascorbate anions are doped into the polymer during the oxidation of the PAN film. Conversely, the oxidation of positively charged DA may only take place at the PAN/solution interface. The different mechanisms of the electrode reactions result in the oxidation of AA and DA occurring at different potentials. As a result, the AuNP/PAN composite electrode can be employed to simultaneously detect AA and DA with a good linear range, high sensitivity, and low detection limit.

  20. Autoradiographic study on the incorporation of carbon-14 labeled formate and adenine into nucleic acid in blood-forming cells

    The incorporation of [14C]formate and [8-14C]adenine into nucleic acid in blood-forming cells was studied by the autoradiographic technique. The isotopic markers were injected subcutaneously into young rats weighting from 100 to 150 g three times every 24 hours and the animals were examined 3 hours after the last injection. In the case of [14C]formate injection, erythroblasts exhibited extremely strong labeling in contrast to weaker labeling of other blood-forming cells. In the case of [14C]adenine administration, on the other hand, immature cells of the granuclocytic series as well as immature reticulum cells (proliferating cells of reticular tissue) were much more heavily labeled than were other blood-forming cells, particularly the erythroblasts which revealed weak or no labeling. By digestion or extraction of DNA, RNA or both from cells with DNase, RNase or hot 10% perchloric acid treatment, respectively, it was confirmed that the observed heavy labeling of any type of cells with either [14C]formate or [14C]adenine was due chiefly to incorporation of the radioactive materials into nuclear DNA. The present results are discussed together with the findings of earlier studies on lymphoid cells which indicate that, in certain cell types, the patterns of [3H]deoxycytidine labeling differ considerably from the corresponding patterns of [3H]deoxycytidine labeling. The present and earlier findings provide evidence to substantiate that, among blood-forming cells, there are considerable variations in the labeling patterns of nuclear DNA depending on differences in the radioactive DNA precursors used as well as in the cell types. (author)

  1. Effect of polyunsaturated fatty acids and phospholipids on ( sup 3 H)-vitamin E incorporation into pulmonary artery endothelial cell membranes

    Sekharam, K.M.; Patel, J.M.; Block, E.R. (Univ. of Florida College of Medicine, Gainesville (USA))

    1990-12-01

    Vitamin E, a dietary antioxidant, is presumed to be incorporated into the lipid bilayer of biological membranes to an extent proportional to the amount of polyunsaturated fatty acids or phospholipids in the membrane. In the present study we evaluated the distribution of incorporated polyunsaturated fatty acids (PUFA) and phosphatidylethanolamine (PE) in various membranes of pulmonary artery endothelial cells. We also studied whether incorporation of PUFA or PE is responsible for increased incorporation of (3H)-vitamin E into the membranes of these cells. Following a 24-hr incubation with linoleic acid (18:2), 18:2 was increased by 6.9-, 9.2-, and 13.2-fold in plasma, mitochondrial, and microsomal membranes, respectively. Incorporation of 18:2 caused significant increases in the unsaturation indexes of mitochondrial and microsomal polyunsaturated fatty acyl chains (P less than .01 versus control in both membranes). Incubation with arachidonic acid (20:4) for 24 hr resulted in 1.5-, 2.3-, and 2.4-fold increases in 20:4 in plasma, mitochondrial, and microsomal membranes, respectively. The unsaturation indexes of polyunsaturated fatty acyl chains of mitochondrial and microsomal membranes also increased (P less than .01 versus control in both membranes). Although incubations with 18:2 or 20:4 resulted in several-fold increases in membrane 18:2 or 20:4 fatty acids, incorporation of (3H)-vitamin E into these membranes was similar to that in controls. Following a 24-hr incubation with PE, membrane PE content was significantly increased, and (3H)-vitamin E incorporation was also increased to a comparable degree, i.e., plasma membrane greater than mitochondria greater than microsomes. Endogenous vitamin E content of the cells was not altered because of increased incorporation of PE and (3H)-vitamin E.

  2. Thermal stability and surface acidity of mesoporous silica doubly doped by incorporation of sulfate and zirconium ions

    A sulfated Si-Zr-MCM-41 solid with highly ordered mesostructure was synthesized through a templated synthesis route where the CTAB surfactant was used as template. During the synthesis procedure, various amounts of (NH4)2SO4 were added into the mixed solution of Zr and Si precursors to in situ sulfate the MCM-41 solids, aiming to enhance the acidity and thermal stability. The resultant materials showed a long-range ordered hexagonal arrangement with high surface area larger than 797 m2/g and an average pore size distributed at approximate 2.5-2.8 nm. The high-resolution TEM observations confirmed that the order of the mesostructure gained when the molar ratio of SO42-/(ZrO2 + SiO2) increased from 0.1 to 0.3 but decreased as it reached 0.5, which is consistent with the results of 29Si MAS-NMR and XRD analysis. Compared to Si-MCM-41, the (Q2 + Q3)/Q4 ratio derived from the NMR spectra of the Zr-doped sample was higher, indicating that zirconium atoms were incorporated into the silica framework. Unexpectedly, in situ sulfation does not enhance the surface Broensted acidity, most likely due to the sulfur retained within the bulk of the materials; however, it indeed improved the thermal stability of the solid and long-range order of the structure

  3. The effect of 1,25-dihydroxycholecalciferol on the multiple forms of alkaline phosphatase and the sialic acid incorporation into microsomes of chick duodenum

    Polyacrylamide disc gel electrophoresis of n-butanol solubilized alkaline phosphatase from chick duodenum revealed that the change of alkaline phosphatase induced by 1,25-(OH)2D3 involved the transformation of desialoenzyme to sialoenzyme. The initial stimulation by 1,25-(OH)2D3 of the incorporation of sialic acid into duodenal microsomes corresponded with the initial increase in calcium absorption. After this initial stimulation, there was a rapid decline in sialic acid incorporation into microsomes decreasing below control levels at 24 hr. Calcium concentration in the microsomes followed a pattern similar to the incorporation of sialic acid into microsomes. The depressed sialic acid incorporation was reversed by the addition of calcium in vitro. These results suggest that the initial action of 1,25-(OH)2D3 is to change the membrane permeability to calcium and to change the subcellular distribution of calcium in the small intestine. The accumulated calcium in the microsomes then stimulates the sialic acid incorporation into desialoenzyme. This results in the changes of isozyme pattern of alkaline phosphatase, viz, the transformation of desialoenzyme to sialoenzyme. The transformed alkaline phosphatase might be one of the factors involved more directly in the regulation of calcium transport in intestine. (auth.)

  4. Sulfuric acid functional zirconium (or aluminum) incorporated mesoporous MCM-48 solid acid catalysts for alkylation of phenol with tert-butyl alcohol

    Several zirconium (or aluminum) incorporated mesoporous MCM-48 solid acid catalysts (SO42−/Zr-MCM-48 and SO42−/Al-MCM-48) were prepared by the impregnation method and their physicochemical properties were characterized by means of XRD, FT-IR, TEM, NH3-TPD and N2 physical adsorption. Also, the catalytic activities of these solid acid catalysts were evaluated by the alkylation of phenol with tert-butyl alcohol. The effect of weight hour space velocity (WHSV), reaction time and reaction temperature on catalytic properties was also studied. The results show that the SO42−/Zr-MCM-48 and SO42−/Al-MCM-48 still have good mesoporous structure and long range ordering. Compared with the Zr (or Al)–MCM-48 samples, SO42−/Zr-MCM-48 and SO42−/Al-MCM-48 solid acid catalysts have strong acidity and exhibit high activities in alkylation reaction of phenol with tert-butyl alcohol. The SO42−/Zr-MCM-48-25 (molar ratio of Si/Zr=0.04) catalyst was found to be the most promising and gave the highest phenol conversion among all catalysts. A maximum phenol conversion of 91.6% with 4-tert-butyl phenol (4-TBP) selectivity of 81.8% was achieved when the molar ratio of tert-butyl alcohol:phenol is 2:1, reaction time is 2 h, the WHSV is 2 h−1 and the reaction temperature is 140 °C. - Highlights: • Sulfuric acid functional mesoporous solid acid catalysts were prepared via impregnation method. • The alkylation of phenol with tert-butyl alcohol was carried out over these solid acid catalysts. • The catalytic activity of SO42−/Zr-MCM-48-25 catalyst is much higher than that of the others. • A maximum phenol conversion of 91.6% was achieved under optimum reaction conditions for SO42−/Zr-MCM-48-25

  5. The effect of zinc and phytic acid on the incorporation of 1-14C-acetate into aflatoxin by resting mycelia of Aspergillus parasiticus

    The effect of zinc and phytic acid on [1-14C]-acetate incorporation into aflatoxins by resting mycelium was studied. When different levels of ZnSO4 were used to study its effect on the incorporation of [1-14C]-acetate into aflatoxins, it was found that the specific radioactivity incorporation into aflatoxins was maximum at the level of 10 mM-ZnSO4. At this concentration the change in the specific radioactivities of aflatoxins B1 + B2 and aflatoxins G1 + G2 were +74.61% and +29.66%, respectively. On the other hand, phytic acid had an inhibitory effect on the incorporation of [1-14C]-acetate. These observations have been correlated in order to find out why soyabean is unable to produce aflatoxins by Aspergillus parasiticus. (orig.)

  6. Reduction of Inflammatory Responses and Enhancement of Extracellular Matrix Formation by Vanillin-Incorporated Poly(Lactic-co-Glycolic Acid) Scaffolds

    Lee, Yujung; Kwon, JeongIl; Khang, Gilson; Lee, Dongwon

    2012-01-01

    Vanillin is one of the major components of vanilla, a commonly used flavoring agent and preservative and is known to exert potent antioxidant and anti-inflammatory activities. In this work, vanillin-incorporated poly(lactic-co-glycolic acid) (PLGA) films and scaffolds were fabricated to evaluate the effects of vanillin on the inflammatory responses and extracellular matrix (ECM) formation in vitro and in vivo. The incorporation of vanillin to PLGA films induced hydrophilic nature, resulting i...

  7. Poloxamer-hydroxyethyl cellulose-α-cyclodextrin supramolecular gels for sustained release of griseofulvin.

    Marcos, Xelhua; Pérez-Casas, Silvia; Llovo, José; Concheiro, Angel; Alvarez-Lorenzo, Carmen

    2016-03-16

    Supramolecular gels of poloxamer-hydroxyethyl cellulose (HEC)-α-cyclodextrin (αCD) were developed aiming to obtain synergisms regarding solubilization and sustained release of griseofulvin for topical application. The effects of αCD concentration (0-10%w/w) on the phase behavior of aqueous dispersions of Pluronic(®) P123 (14%w/w) mixed with HEC (2%w/w) were evaluated at 4, 20 and 37°C. The cooperative effects of the inclusion complex formation between poly(ethylene oxide) (PEO) blocks and HEC with αCD prevented phase separation and led to supramolecular networks that solubilize the antifungal drug. Rheological and bioadhesive properties of gels with and without griseofulvin could be easily tuned modulating the polymers proportions. Supramolecular gels underwent sol-gel transition at lower temperature than P123 solely dispersions and enabled drug sustained release for at least three weeks. All gels demonstrated good biocompatibility in the HET-CAM test. Furthermore, the drug-loaded gels showed activity against Trichophyton rubrum and Trichophyton mentagrophytes and thus may be useful for the treatment of tinea capitis and other cutaneous fungal infections. PMID:26795192

  8. Membrane Sealant Poloxamer P188 Protects Against Isoproterenol Induced Cardiomyopathy in Dystrophin Deficient Mice

    Sali Arpana

    2011-05-01

    Full Text Available Abstract Background Cardiomyopathy in Duchenne muscular dystrophy (DMD is an increasing cause of death in patients. The absence of dystrophin leads to loss of membrane integrity, cell death and fibrosis in cardiac muscle. Treatment of cardiomyocyte membrane instability could help prevent cardiomyopathy. Methods Three month old female mdx mice were exposed to the β1 receptor agonist isoproterenol subcutaneously and treated with the non-ionic tri-block copolymer Poloxamer P188 (P188 (460 mg/kg/dose i.p. daily. Cardiac function was assessed using high frequency echocardiography. Tissue was evaluated with Evans Blue Dye (EBD and picrosirius red staining. Results BL10 control mice tolerated 30 mg/kg/day of isoproterenol for 4 weeks while death occurred in mdx mice at 30, 15, 10, 5 and 1 mg/kg/day within 24 hours. Mdx mice tolerated a low dose of 0.5 mg/kg/day. Isoproterenol exposed mdx mice showed significantly increased heart rates (p Conclusions This model suggests that chronic intermittent intraperitoneal P188 treatment can prevent isoproterenol induced cardiomyopathy in dystrophin deficient mdx mice.

  9. Role of leucine in isoprenoid metabolism. Incorporation of (3-/sup 13/C)leucine and of (2-/sup 3/H,4-/sup 14/C)-. beta. ,. beta. -dimethyl-acrylic acid into phytosterols by tissue cultures of Andrographis paniculata

    Anastasis, P.; Freer, I.; Overton, K.; Rycroft, D.; Singh, S.B. (Glasgow Univ. (UK). Dept. of Chemistry)

    1985-02-01

    (3-/sup 13/C)Leucine is incorporated into phytosterols by tissue cultures of Andrographis paniculata by breakdown to acetyl-CoA and its subsequent incorporation via (3S)-3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) and mevalonic acid; (2-/sup 3/H,4-/sup 14/C)-..beta..,..beta..-dimethylacrylic acid also is not incorporated intact.

  10. 5-aminolevulinic acid-incorporated poly(vinyl alcohol nanofiber-coated metal stent for application in photodynamic therapy

    Kang DH

    2012-05-01

    Full Text Available Jin Ju Yoo*, Chan Kim*, Chung-Wook Chung, Young-Il Jeong, Dae Hwan KangNational Research and Development Center for Hepatibiliary Disease, Pusan National University Yangsan Hospital, Gyeongnam 626-770, Korea *These authors contributed equally to this work.Background: The study investigated the use of combined photodynamic therapy (PDT and stent placement for the treatment of cholangiocarcinoma (CC. For this purpose, 5-aminolevulinic acid (ALA was incorporated into poly(vinyl alcohol (PVA nanofiber, and coated onto metal stents. Their efficacy was assessed in PDT towards HuCC-T1 CC cells.Methods: Fabrication of ALA-PVA nanofiber, and simultaneous coating onto metal stents, was performed through electrospinning. The dark-toxicity, generation of protoporphyrin IX (PpIX, and PDT effect of ALA and ALA-PVA nanofiber were studied in vitro, using HuCC-T1 CC cells.Results: The ALA-PVA nanofibers were coated onto metal stents less than 1000 nm in diameter. ALA-only displayed marginal cytotoxicity; ALA-PVA nanofiber showed less cytotoxicity. PpIX generation was not sigficantly different between ALA and ALA-PVA nanofiber treatments. PVA itself did not generate PpIX in tumor cells. ALA and ALA-PVA nanofiber displayed a similar PDT effect on tumor cells. Cell viability was decreased, dose-dependently, until ALA concentration reached 100 µg/mL. Necrosis and apoptosis of tumor cells occurred similarly for ALA and ALA- PVA nanofiber treatments.Conclusion: The ALA-PVA nanofiber-coated stent is a promising candidate for therapeutic use with cholangiocarcinoma.Keywords: nanofiber, photodynamic therapy, 5-aminolevulinic acid, poly(vinyl alcohol

  11. Fabrication of phosphotungstic acid functionalized mesoporous silica composite membrane by alternative tape-casting incorporating phase inversion technique

    Bai, Li; Zhang, Lan; He, Hong Quan; Rasheed, Raj Kamal S./O. Abdul; Zhang, Cai Zhi; Ding, Ovi Lian; Chan, Siew Hwa

    2014-01-01

    Meso-porous silica (MCM-41) membranes functionalized by phosphotungstic acid (HPW) for high temperature proton exchange membrane fuel cells (HT-PEMFCs) are successfully developed by a cost-effective tape-casting incorporating phase inversion and vacuum assisted wet impregnation techniques. The microstructure of the membrane is characterized by field emission scanning electron microscopy (FESEM). The effect of MCM-41 content on the tensile strength, ultimate elongation, and weight gain ratio and swelling ratio in water/methanol of the membranes are investigated in detail. The thermal stability of MCM-41 membrane with/without HPW is analyzed by thermogravimetric analysis (TGA) and derivative thermogravimetry (DTG) techniques. In particular, the effects of HPW loading and MCM-41 content on the proton conductivity of HPW/MCM-41 membranes are studied comprehensively. The results on the swelling ratio and tensile tension show that the developed membranes can be applied as an electrolyte membrane for HT-PEMFCs. The developed MCM-41 membrane, in which polyethersulfone (PESf) is used as the supporting backbone, is able to operate up to 200 °C. The single cell assembled from HPW/MCM-41 membrane with 70 wt.% HPW loading gives a peak output power of ∼230 mW cm-2 and ∼125 mW cm-2 in H2/air at 90 °C and in methanol/air at 150 °C without any humidification, respectively.

  12. Incorporation of eicosapentaenoic and docosahexaenoic acids into lipid pools when given as supplements providing doses equivalent to typical intakes of oily fish 1 2 3 4

    Browning, Lucy M; Walker, Celia G.; Mander, Adrian P; West, Annette L.; Madden, Jackie; Gambell, Joanna M; Young, Stephen; Wang, Laura; Jebb, Susan A.; Calder, Philip C.

    2012-01-01

    Background: Estimation of the intake of oily fish at a population level is difficult. The measurement of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in biological samples may provide a useful biomarker of intake. Objective: We identified the most appropriate biomarkers for the assessment of habitual oily fish intake and changes in intake by elucidating the dose- and time-dependent response of EPA and DHA incorporation into various biological samples that represent roles in fatt...

  13. Hydrogen exchange during cell-free incorporation of deuterated amino acids and an approach to its inhibition

    Perdeuteration, selective deuteration, and stereo array isotope labeling (SAIL) are valuable strategies for NMR studies of larger proteins and membrane proteins. To minimize scrambling of the label, it is best to use cell-free methods to prepare selectively labeled proteins. However, when proteins are prepared from deuterated amino acids by cell-free translation in H2O, exchange reactions can lead to contamination of 2H sites by 1H from the solvent. Examination of a sample of SAIL-chlorella ubiquitin prepared by Escherichia coli cell-free synthesis revealed that exchange had occurred at several residues (mainly at Gly, Ala, Asp, Asn, Glu, and Gln). We present results from a study aimed at identifying the exchanging sites and level of exchange and at testing a strategy for minimizing 1H contamination during wheat germ cell-free translation of proteins produced from deuterated amino acids by adding known inhibitors of transaminases (1 mM aminooxyacetic acid) and glutamate synthetase (0.1 mM l-methionine sulfoximine). By using a wheat germ cell-free expression system, we produced [U–2H, 15N]-chlorella ubiquitin without and with added inhibitors, and [U–15N]-chlorella ubiquitin as a reference to determine the extent of deuterium incorporation. We also prepared a sample of [U–13C, 15N]-chlorella ubiquitin, for use in assigning the sites of exchange. The added inhibitors did not reduce the protein yield and were successful in blocking hydrogen exchange at Cα sites, with the exception of Gly, and at Cβ sites of Ala. We discovered, in addition, that partial exchange occurred with or without the inhibitors at certain side-chain methyl and methylene groups: Asn–Hβ, Asp–Hβ, Gln–Hγ, Glu–Hγ, and Lys–Hε. The side-chain labeling pattern, in particular the mixed chiral labeling resulting from partial exchange at certain sites, should be of interest in studies of large proteins, protein complexes, and membrane proteins.

  14. Hydrogen exchange during cell-free incorporation of deuterated amino acids and an approach to its inhibition

    Tonelli, Marco; Singarapu, Kiran K. [University of Wisconsin-Madison, National Magnetic Resonance Facility at Madison (NMRFAM), Department of Biochemistry (United States); Makino, Shin-ichi; Sahu, Sarata C.; Matsubara, Yuko [University of Wisconsin-Madison, Center for Eukaryotic Structural Genomics (CESG), Department of Biochemistry (United States); Endo, Yaeta [Ehime University, Cell-Free Science and Technology Research Center (Japan); Kainosho, Masatsune [Tokyo Metropolitan University, Center for Priority Areas (Japan); Markley, John L., E-mail: markley@nmrfam.wisc.edu [University of Wisconsin-Madison, National Magnetic Resonance Facility at Madison (NMRFAM), Department of Biochemistry (United States)

    2011-12-15

    Perdeuteration, selective deuteration, and stereo array isotope labeling (SAIL) are valuable strategies for NMR studies of larger proteins and membrane proteins. To minimize scrambling of the label, it is best to use cell-free methods to prepare selectively labeled proteins. However, when proteins are prepared from deuterated amino acids by cell-free translation in H{sub 2}O, exchange reactions can lead to contamination of {sup 2}H sites by {sup 1}H from the solvent. Examination of a sample of SAIL-chlorella ubiquitin prepared by Escherichia coli cell-free synthesis revealed that exchange had occurred at several residues (mainly at Gly, Ala, Asp, Asn, Glu, and Gln). We present results from a study aimed at identifying the exchanging sites and level of exchange and at testing a strategy for minimizing {sup 1}H contamination during wheat germ cell-free translation of proteins produced from deuterated amino acids by adding known inhibitors of transaminases (1 mM aminooxyacetic acid) and glutamate synthetase (0.1 mM l-methionine sulfoximine). By using a wheat germ cell-free expression system, we produced [U-{sup 2}H, {sup 15}N]-chlorella ubiquitin without and with added inhibitors, and [U-{sup 15}N]-chlorella ubiquitin as a reference to determine the extent of deuterium incorporation. We also prepared a sample of [U-{sup 13}C, {sup 15}N]-chlorella ubiquitin, for use in assigning the sites of exchange. The added inhibitors did not reduce the protein yield and were successful in blocking hydrogen exchange at C{sup {alpha}} sites, with the exception of Gly, and at C{sup {beta}} sites of Ala. We discovered, in addition, that partial exchange occurred with or without the inhibitors at certain side-chain methyl and methylene groups: Asn-H{sup {beta}}, Asp-H{sup {beta}}, Gln-H{sup {gamma}}, Glu-H{sup {gamma}}, and Lys-H{sup {epsilon}}. The side-chain labeling pattern, in particular the mixed chiral labeling resulting from partial exchange at certain sites, should be of

  15. A randomized study of the effect of fish oil on n-3 fatty acid incorporation and nutritional status in lung cancer patients

    Andersen, Jens Rikardt; Dannerfjord, Stina Hjerrild; Nørgaard, Michael; Lauritzen, Lotte; Lange, Peter; Jensen, N Aa; Boisen, L W; Jensen, R G; Andersen, M J; Sørensen, J B

    2015-01-01

    Long-chain n-3 polyunsaturated fatty acids (n-3 LCPUFA) have been proposed to have beneficial effect on cancer cachexia. The aims of the present study were to a) determine the incorporation of n-3 LCPUFA in erythrocytes (RBC) as a measurement of compliance to fish oil (FO)-supplement in lung cancer...

  16. Modulation of the rheological and mucoadhesive properties of thermosensitive poloxamer-based hydrogels intended for the rectal administration of quinine.

    Koffi, A A; Agnely, F; Ponchel, G; Grossiord, J L

    2006-03-01

    The aim of this work was to formulate and characterize thermosensitive gels based on poloxamer 407, a thermosensitive polymer, and hydroxypropylmethylcellulose (HPMC), a bioadhesive polymer, intended for the rectal delivery of quinine in children. In order to avoid the macroscopic phase separation between the two polymers it was necessary to add propanediol-1,2. After the formulation of stable ternary systems, their rheological properties were studied as a function of temperature. It was thus possible to obtained the gelation temperature of the systems as well as their elastic modulus, G', at 37 degrees C. It appeared that HPMC in the presence of propanediol-1,2 had a synergistic effect on the gelation of poloxamer 407. Tests on the rabbit rectal mucous membrane, founded on a technique of traction of the adhesive/adhered joint, made it possible to characterize the bioadhesive properties of the gels by measuring the work of adhesion, W, and the maximum detachment force, F(max). Over small concentration ranges of poloxamer and HPMC, very important variations of the viscoelastic and mucoadhesives properties were observed. It was shown that the viscoelastic and the bioadhesive properties were tightly correlated. Indeed, an empirical equivalence relation was established between the shear frequency and the rate of deformation in traction and allowed to describe all our samples by a master curve. This master curve would make it possible to predict the values of W at various rates of traction by the simple non-destructive measurement of the elastic modulus, G', at an equivalent shear frequency. PMID:16356700

  17. Oxygen-18 incorporation into malic acid during nocturnal carbon dioxide fixation in crassulacean acid metabolism plants: a new approach to estimating in vivo carbonic anhydrase activity

    Holtum, J.A.M.; Summons, R.; Roeske, C.A.; Comins, H.N.; O' Leary, M.H.

    1984-01-01

    Crassulacean acid metabolism (CAM) plants fix carbon dioxide at night by the carboxylation of phosphoenolpyruvate. If CO2 fixation is conducted with TC YO2, then in the absence of carbonic anhydrase, the malate formed by dark CO2 fixation should also contain high levels of carbon-13 and oxygen-18. Conversely, if carbonic anhydrase is present and highly active, oxygen exchange between CO2 and cellular H2O will occur more rapidly than carboxylation, and the ( TC) malate formed will contain little or no oxygen-18 above the natural abundance level. The presence of oxygen-18 in these molecules can be detected either by nuclear magnetic resonance or by mass spectrometry. Studies of phosphoenolpyruvate carboxylase in the presence and absence of carbonic anhydrase in vitro confirm the validity of the method. When CAM plants are studied by this method, we find that most species show incorporation of a significant amount of oxygen-18. Comparison of these results with results of isotope fractionation and gas exchange studies permits calculation of the in vivo activity of carbonic anhydrase toward HCO3 compared with that of phosphoenolpyruvate carboxylase. The ratio (carbonic anhydrase activity/phosphoenolpyruvate carboxylase activity) is species dependent and varies from a low of about 7 for Ananas comosus to values near 20 for Hoya carnosa and Bryophyllum pinnatum, 40 for Kalanchoee daigremontiana, and 100 or greater for Bryophyllum tubiflorum, Kalanchoee serrata, and Kalanchoae tomentosa. Carbonic anhydrase activity increases relative to phosphoenolpyruvate carboxylase activity at higher temperature. 37 references, 2 figures, 8 tables.

  18. Profiling of Polar Lipids in Marine Oleaginous Diatom Fistulifera solaris JPCC DA0580: Prediction of the Potential Mechanism for Eicosapentaenoic Acid-Incorporation into Triacylglycerol

    Yue Liang

    2014-05-01

    Full Text Available The marine oleaginous diatom Fistulifera solaris JPCC DA0580 is a candidate for biodiesel production because of its high lipid productivity. However, the substantial eicosapentaenoic acid (EPA content in this strain would affect the biodiesel quality. On the other hand, EPA is also known as the essential health supplement for humans. EPAs are mainly incorporated into glycerolipids in the microalgal cell instead of the presence as free fatty acids. Therefore, the understanding of the EPA biosynthesis including the incorporation of the EPA into glycerolipids especially triacylglycerol (TAG is fundamental for regulating EPA content for different purposes. In this study, in order to identify the biosynthesis pathway for the EPA-containing TAG species, a lipidomic characterization of the EPA-enriched polar lipids was performed by using direct infusion electrospray ionization (ESI-Q-TRAP-MS and MS/MS analyses. The determination of the fatty acid positional distribution showed that the sn-2 position of all the chloroplast lipids and part of phosphatidylcholine (PC species was occupied by C16 fatty acids. This result suggested the critical role of the chloroplast on the lipid synthesis in F. solaris. Furthermore, the exclusive presence of C18 fatty acids in PC highly indicated the biosynthesis of EPA on PC. Finally, the PC-based acyl-editing and head group exchange processes were proposed to be essential for the incorporation of EPA into TAG and chloroplast lipids.

  19. Structural and Functional Recovery of Electropermeabilized Skeletal Muscle in-vivo after Treatment with Surfactant Poloxamer 188

    Collins, John M.; Despa, Florin; Lee, Raphael C.

    2007-01-01

    A critical requirement for cell survival after trauma is sealing of breaks in the cell membrane [1–11]. The triblock copolymer surfactant Poloxamer 188 (P188) is known to increase the cell survival after membrane electroporation [8,22]. Here, we use a rat hind-limb model of electroporation injury to determine if the intravenous administration of P188 improves the recovery of the muscle function. Rat hind-limbs received a sequence of either 0, 3, 6, 9, or 12 electrical current pulses (2 ampere...

  20. Randomized Controlled Trial Examining the Effects of Fish Oil and Multivitamin Supplementation on the Incorporation of n-3 and n-6 Fatty Acids into Red Blood Cells

    Andrew Pipingas

    2014-05-01

    Full Text Available The present randomized, placebo-controlled, double-blind, parallel-groups clinical trial examined the effects of fish oil and multivitamin supplementation on the incorporation of n-3 and n-6 fatty acids into red blood cells. Healthy adult humans (n = 160 were randomized to receive 6 g of fish oil, 6 g of fish oil plus a multivitamin, 3 g of fish oil plus a multivitamin or a placebo daily for 16 weeks. Treatment with 6 g of fish oil, with or without a daily multivitamin, led to higher eicosapentaenoic acid (EPA composition at endpoint. Docosahexaenoic acid (DHA composition was unchanged following treatment. The long chain LC n-3 PUFA index was only higher, compared to placebo, in the group receiving the combination of 6 g of fish oil and the multivitamin. Analysis by gender revealed that all treatments increased EPA incorporation in females while, in males, EPA was only significantly increased by the 6 g fish oil multivitamin combination. There was considerable individual variability in the red blood cell incorporation of EPA and DHA at endpoint. Gender contributed to a large proportion of this variability with females generally showing higher LC n-3 PUFA composition at endpoint. In conclusion, the incorporation of LC n-3 PUFA into red blood cells was influenced by dosage, the concurrent intake of vitamin/minerals and gender.

  1. Changes in soil chemical properties and lettuce yield response following incorporation of biochar and cow dung to highly weathered acidic soils

    Agyei Frimpong, Kwame; Amoakwah, Emmanuel; Osei, Benjamin;

    2016-01-01

    imposed on two highly weathered, acidic soils from the coastal savanna and tropical rainforest agroecological zones of Ghana, respectively, to elucidate their effect on yield of lettuce. The study showed that application of biochar solely or in combination with cow dung increased soil pH, total organic...... carbon, and cation exchange capacity, and temporarily increased soil respiration and microbial biomass carbon. Further, incorporation of combined application of cow dung and biochar increased lettuce yield more than sole incorporation of either amendment. The study demonstrated that corn cob biochar can...... improve soil chemical properties and lettuce yield if applied solely or in combination with cow dung....

  2. INCORPORATION OF OMEGA-3 FATTY ACID AND SYNTHESIS OF THEIR METABOLITES IN MUSCLE TISSUE IN MICE FED WITH FLAXSEED AND PERILLA-ENRICHED DIET

    Sheisa Cyleia Sargi

    2013-01-01

    Full Text Available Polyunsaturated fatty acids omega 3 and omega 6 are believed to have beneficial physiological effects on the human body. The present study aimed to evaluate the incorporation and synthesis of omega-3 fatty acid and their metabolites in muscle tissue in mice fed with a flaxseed and perilla-enriched diet. The animals were fed a diet supplemented with flaxseed and perilla meals for fifty-six days and control animals received a commercial diet. Mice were sacrificed on the 7th, 28th and 56th days and the muscle tissue was collected. The FA concentration was analyzed by gas chromatography. The levels of alpha-Linolenic Acid (LNA and their metabolites, Eicosapentaenoic Acid (EPA and Docosahexaenoic Acid (DHA increased as a function of feeding length in the groups given the supplemented diet. LNA was incorporated into muscle tissue and conversion into Very Long Chain Polyunsaturated Fatty Acids (VLC-PUFA was observed. The muscle tissue of animals fed with enriched diets presented a high sum of n-3 FA and high concentrations of LNA, EPA and DHA, as compared to mice fed commercial diet. The intake of flaxseed and perilla-meal enriched diets by mice may result in the deposition of LNA and its metabolites EPA and DHA into muscle tissue.

  3. Poloxamer 407-based intranasal thermoreversible gel of zolmitriptan-loaded nanoethosomes: formulation, optimization, evaluation and permeation studies.

    Shelke, Santosh; Shahi, Sadhana; Jalalpure, Sunil; Dhamecha, Dinesh

    2016-12-01

    Zolmitriptan is the drug of choice for migraine, but low oral bioavailability (ethosomes was optimized by using 3(2) factorial design for percent drug entrapment efficiency, vesicle size, zeta potential, and polydispersity index. Optimized formulation E6 showed the vesicle size (171.67 nm) and entrapment efficiency (66%) when compared with the other formulations. Thermoreversible gels prepared by using poloxamer 407 showed the phase transition temperature at 32-33 °C which was in line with the nasal physiological temperature. The optimized ethosomes were loaded into the thermoreversible mucoadhesive gel optimized by varying concentrations of poloxamer 407, carbopol 934, HPMC K100, and evaluated for gel strength, gelation temperature, mucoadhesive strength, in vitro drug release, and ex vivo drug permeation, where G3 and G6 were found to be optimized formulations. In vitro drug release was studied by different kinetic models suggested that G3 (n = 0.582) and G6 (n = 0.648) showed Korsemeyer-Peppas (KKP) model indicating non-Fickian release profiles. A permeation coefficient of 5.92 and 5.9 µg/cm(2) for G3 and G6, respectively, revealed very little difference in release rate after 24 h between both the formulations. Non-toxic nature of the gels on columnar epithelial cells was confirmed by histopathological evaluation. PMID:26758957

  4. IR 820 dye encapsulated in polycaprolactone glycol chitosan: Poloxamer blend nanoparticles for photo immunotherapy for breast cancer

    Kumar, Piyush; Srivastava, Rohit, E-mail: rsrivasta@iitb.ac.in

    2015-12-01

    In the present study, we have fabricated biocompatible and biodegradable monodisperse IR 820 encapsulated polycaprolactone (PCL) glycol chitosan (GC): Poloxamer blend nanoparticles (PP-IR NPs) for imaging and effective photo-immunotherapy. IR 820 has been used as an imaging and photothermal agent whereas glycol chitosan (GC) as an immunostimulatory agent. The combination of IR 820, poloxamer, and GC can be used effectively for photoimmunotherapy for cancer, drug-resistant and TNF-α resistant estrogen positive breast cancer. PP-IR NPs are stable in aqueous solution. The uniform size of 100–220 nm with a high zeta value of + 38 ± 2 mV led them to accumulate in cancer cells. Laser treatment did not affect the morphology of PP-IR NPs as observed under the transmission electron microscope (TEM). In vitro cytotoxicity studies on MCF-7 cells showed enhanced toxicity upon laser treatment. Further, we validated the cell death by reactive oxygen species (ROS) production. Our studies thus showed that PP-IR NPs are effective in suppressing metastatic cancer as the combinational therapy leads to the formation of apoptotic bodies in MCF-7 cells. - Highlights: • PPIR nanoparticles for photoimmunotherapy for cancer • IR 820/GC serves as theranostic and immunostimulatory. • Photoimmunotherapy enhances cytotoxicity by reactive oxygen species production.

  5. IR 820 dye encapsulated in polycaprolactone glycol chitosan: Poloxamer blend nanoparticles for photo immunotherapy for breast cancer

    In the present study, we have fabricated biocompatible and biodegradable monodisperse IR 820 encapsulated polycaprolactone (PCL) glycol chitosan (GC): Poloxamer blend nanoparticles (PP-IR NPs) for imaging and effective photo-immunotherapy. IR 820 has been used as an imaging and photothermal agent whereas glycol chitosan (GC) as an immunostimulatory agent. The combination of IR 820, poloxamer, and GC can be used effectively for photoimmunotherapy for cancer, drug-resistant and TNF-α resistant estrogen positive breast cancer. PP-IR NPs are stable in aqueous solution. The uniform size of 100–220 nm with a high zeta value of + 38 ± 2 mV led them to accumulate in cancer cells. Laser treatment did not affect the morphology of PP-IR NPs as observed under the transmission electron microscope (TEM). In vitro cytotoxicity studies on MCF-7 cells showed enhanced toxicity upon laser treatment. Further, we validated the cell death by reactive oxygen species (ROS) production. Our studies thus showed that PP-IR NPs are effective in suppressing metastatic cancer as the combinational therapy leads to the formation of apoptotic bodies in MCF-7 cells. - Highlights: • PPIR nanoparticles for photoimmunotherapy for cancer • IR 820/GC serves as theranostic and immunostimulatory. • Photoimmunotherapy enhances cytotoxicity by reactive oxygen species production

  6. Enhancement in bioavailability of ketorolac tromethamine via intranasal in situ hydrogel based on poloxamer 407 and carrageenan.

    Li, Chenxi; Li, Chunyan; Liu, Zheshuo; Li, Qiuhong; Yan, Xueying; Liu, Yu; Lu, Weiyue

    2014-10-20

    The objective of this study was to construct a new in situ gel system based on the combination of poloxamer 407 and carrageenan (carrageenan-poloxamer 407 hydrogel, CPH) for intranasal delivery of ketorolac tromethamine. CPH showed potassium ion concentration - dependent erosion characteristics which ensured slow erosion in aqueous environment containing potassium ion at the physiological level. Loading with ketorolac tromethamine influenced erosion, drug release and thermosensitive properties of CPH. CPH containing 15% ketorolac tromethamine showed suitable gelation temperature (near 35°C) and in vitro sustained release profiles. Pharmacokinetic study of intranasal CPH containing 15% ketorolac tromethamine in rats demonstrated enhanced absolute bioavailability (68.8 ± 23.3%) and prolonged mean residence time (8.8 ± 3.5h) in comparison with the intranasal solution group (24.8 ± 13.8%, 3.9 ± 0.6h). Nasal ciliotoxicity evaluation on an in situ toad palate model preliminarily showed the safety of CPH for intranasal use. All results suggested the potential of CPH as a new sustained - release platform for the intranasal delivery of ketorolac tromethamine. PMID:25138250

  7. Gas chromatographic measurement of 3- and 4-thia fatty acids incorporated into various classes of rat liver lipids during feeding experiments.

    Grav, H J; Asiedu, D K; Berge, R K

    1994-08-01

    A practical procedure is described for the quantitative measurement of the amount of acyl units derived from tetradecylthioacetic acid (effecting hypolipemia in rats) and tetradecylthiopropionic acid (effecting hyperlipidemia). The procedure involves three main successive steps: (1) extraction; (2) solid-phase lipid class separation yielding free fatty acids, phospholipids, triacylglycerides, cholesterol esters, and diacylglycerides without crosscontamination; and (3) gas chromatography of hydrolyzed lipids derivatized to picolinyl esters, combined with unambiguous identification by gas chromatography-mass spectrometry. The overall recoveries of heptadecanoyl lipids added as internal standards during extraction were 94-96%, except for cholesteryl heptadecanoate where the recovery was 60% owing to incomplete hydrolysis. Recoveries of thia fatty acids from samples spiked with these compounds were 95%. Flame-ionization response factors were found to be 0.92 and 0.81 for the tetradecylthioacetic acid and tetradecylthiopropionic acid picolinyl esters, respectively, compared to that of heptadecanoic acid. The lower limit of quantitation was 25 pmol as injected. Measurement of the amount of thia fatty acyl units in rat plasma and in liver lipids 4 h after administration of single doses by gastric intubation indicated efficient absorbtion and rapid incorporation into liver lipids, particularly in the phospholipid fraction. Both plasma clearance and channelling into lipids was slower for the 4-thia fatty acid. PMID:7952108

  8. Poloxamer 188 and antioxidants prevent acute radiation necrosis of adult skeletal muscle cells

    Full text: To date there are no effective therapeutic agents widely available for acute radiation sickness. Acute cellular necrosis occurring minutes to hours after exposure to high doses of ionizing radiation (IR) results from rapid membrane lipid peroxidation, blebbing and membrane breakdown. Not only can repairing the membrane prevent acute necrosis, but it can also provide a critical time period to address other mechanisms of cell death, such as apoptosis or mitotic arrest, which manifest over a longer time frame. We have previously shown that certain polymer surfactants can restore structural integrity and transport barrier function of cell membranes following high-dose IR. We now present data showing that the amphiphillic surfactant Poloxamer 188 (P188), a tri-block copolymer composed of two hydrocephalic blocks separated by a hydrophobic central block, has efficacy in preventing acute necrosis of adult rat skeletal muscle cells after high-dose IR and that at doses in which P188 is effective, adding the antioxidant ascorbate or n-acetyl cysteine further increases cell survival. Explanted rat flexor digitoum brevis muscle cells received 10, 40 or 40 Gy IR from Co 60 in a 21% oxygen atmosphere and their viability was determined using fluorometric probes (Calcein-AM and Ethidium homodimer) at 4 and 18 hours after IR. Compared to unirradiated cells, 10 Gy did not cause acute necrosis. Significant acute necrosis was observed after 40 and 80 Gy in a dose-dependent manner. Post-IR treatment with P188 significantly enhanced the cells' viability. By comparison, treating with 10 kDa neutral Dextran, a purely hydrophilic polymer, was not found to be effective. Despite progressive cell death over 18 h after high-dose IR, cells treated with P188 showed greater survival than cells grown only in media or Dextran-treated cells. Cells treated with 40 Gy survived better than those treated with 80 Gy, indicating some limits to the efficacy of treatment with P188. Cells

  9. Synthesis of unusual alpha-amino acids and study of the effect of their incorporation into antimicrobial peptides. Total synthesis of biactive marine natural products and analogues thereof

    El Marrouni El Ghazaoui, Abdellatif

    2012-01-01

    The principle theme of this thesis was the synthesis of bioactive compounds. To this end, this work was focus on two main projects. The first one, which was carried out in the Department of Chemistry of the University of Girona under the supervision of Dr Montserrat Heras, concerned the synthesis of new unnatural amino acids bearing a pyrimidine ring within their side chain for incorporation into the antimicrobial peptide BP100 following a rational design in order to improve its biological pr...

  10. Eicosapentaenoic acid membrane incorporation impairs ABCA1-dependent cholesterol efflux via a protein kinase A signaling pathway in primary human macrophages.

    Fournier, Natalie; Tardivel, Sylviane; Benoist, Jean-François; Vedie, Benoît; Rousseau-Ralliard, Delphine; Nowak, Maxime; Allaoui, Fatima; Paul, Jean-Louis

    2016-04-01

    A diet rich in n-3/n-6 polyunsaturated fatty acids (PUFAs) is cardioprotective. Dietary PUFAs affect the cellular phospholipids composition, which may influence the function of membrane proteins. We investigated the impact of the membrane incorporation of several PUFAs on ABCA1-mediated cholesterol efflux, a key antiatherogenic pathway. Arachidonic acid (AA) (C20:4 n-6) and docosahexaenoic acid (DHA) (C22:6 n-3) decreased or increased cholesterol efflux from J774 mouse macrophages, respectively, whereas they had no effect on efflux from human monocyte-derived macrophages (HMDM). Importantly, eicosapentaenoic acid (EPA) (C20:5 n-3) induced a dose-dependent reduction of ABCA1 functionality in both cellular models (-28% for 70μM of EPA in HMDM), without any alterations in ABCA1 expression. These results show that PUFA membrane incorporation does not have the same consequences on cholesterol efflux from mouse and human macrophages. The EPA-treated HMDM exhibited strong phospholipid composition changes, with high levels of both EPA and its elongation product docosapentaenoic acid (DPA) (C22:5 n-3), which is associated with a decreased level of AA. In HMDM, EPA reduced the ATPase activity of the membrane transporter. Moreover, the activation of adenylate cyclase by forskolin and the inhibition of cAMP phosphodiesterase by isobutylmethylxanthine restored ABCA1 cholesterol efflux in EPA-treated human macrophages. In conclusion, EPA membrane incorporation reduces ABCA1 functionality in mouse macrophages as well as in primary human macrophages and this effect seems to be PKA-dependent in human macrophages. PMID:26776055

  11. Different kinetic in incorporation and depletion of n-3 fatty acids in erythrocytes and leukocytes of mice

    Mu, Huiling; Thogersen, Regitze Louise; Maaetoft-Udsen, Kristina;

    2006-01-01

    during a 6-wk feeding period. Over the first 3-wk period (the incorporation period) the mice were fed a special diet with a high n-3/n-6 PUFA ratio. In the following 3-wk period (the depletion period) the mice were fed a standard chow diet. A linear increase of the concentration of EPA and DHA in...... erythrocyte membranes was observed during the incorporation period, whereas a stagnation was observed after the second week for leukocytes. The level of EPA did not fall to the background level after the depletion period, and the level of DHA was kept almost constant during the depletion period in the...... erythrocyte membranes. In leukocytes the concentration of both EPA and DHA decreased during the depletion period, but did not reach the background level after the 3-wk depletion. In conclusion, the kinetics of EPA and DHA in the different cells are different. The rate of incorporation is faster than that of...

  12. The chain length of lignan macromolecule from flaxseed hulls is determined by the incorporation of coumaric acid glucosides and ferulic acid glucosides

    Struijs, K.; Vincken, J.P.; Doeswijk, T.G.; Voragen, A.G.J.; Gruppen, H.

    2009-01-01

    Lignan macromolecule from flaxseed hulls is composed of secoisolariciresinol diglucoside (SDG) and herbacetin diglucoside (HDG) moieties ester-linked by 3-hydroxy-3-methylglutaric acid (HMGA), and of p-coumaric acid glucoside (CouAG) and ferulic acid glucoside (FeAG) moieties ester-linked directly t

  13. Optimized Lentiviral Transduction Protocols by Use of a Poloxamer Enhancer, Spinoculation, and scFv-Antibody Fusions to VSV-G.

    Anastasov, Nataša; Höfig, Ines; Mall, Sabine; Krackhardt, Angela M; Thirion, Christian

    2016-01-01

    Lentiviral vectors (LV) are widely used to successfully transduce cells for research and clinical applications. This optimized LV infection protocol includes a nontoxic poloxamer-based adjuvant combined with antibody-retargeted lentiviral particles. The novel poloxamer P338 demonstrates superior characteristics for enhancing lentiviral transduction over the best-in-class polybrene-assisted transduction. Poloxamer P338 exhibited dual benefits of low toxicity and high efficiency of lentiviral gene delivery into a range of different primary cell cultures. One of the major advantages of P338 is its availability in pharma grade and applicability as cell culture medium additive in clinical protocols. Lentiviral vectors pseudotyped with the vesicular stomatitis virus glycoprotein (VSV-G) can be produced to high titers and mediate high transduction efficiencies in vitro. For clinical applications the need for optimized transduction protocols, especially for transduction of primary T and stem cells, is high. The successful use of retronectin, the second lentivirus enhancer available as GMP material, requires the application of specific coating protocols not applicable in all processes, and results in the need of a relatively high multiplicity of infection (MOI) to achieve effective transduction efficiencies for hematopoietic cells (e.g., CD34+ hematopoietic stem cells). Cell specificity of lentiviral vectors was successfully increased by displaying different ratios of scFv-fused VSV-G glycoproteins on the viral envelope. The system has been validated with human CD30+ lymphoma cells, resulting in preferential gene delivery to CD30+ cells, which was increased fourfold in mixed cell cultures, by presenting scFv antibody fragments binding to respective surface markers. A combination of spinoculation and poloxamer-based chemical adjuvant increases the transduction of primary T-cells by greater than twofold. The combination of poloxamer-based and scFv-retargeted LVs increased

  14. Comparison of Isotope-labeled Amino Acid Incorporation Rates (CILAIR) Provides a Quantitative Method to Study Tissue Secretomes

    Roelofsen, Johan; Dijkstra, Martijn; Weening, Desiree; de Vries, Marcel; Hoek, Annemieke; Vonk, Roel J.

    2009-01-01

    Adipose tissue is an endocrine organ involved in regulation of whole-body energy metabolism via storage of lipids and secretion of various peptide hormones (adipokines). We previously characterized the adipose tissue secretome and showed that [(13)C]lysine incorporation into secreted proteins can be

  15. Evaluation of the environmental implications of the incorporation of feed-use amino acids in the manufacturing of pig and broiler feeds using Life Cycle Assessment.

    Mosnier, E; van der Werf, H M G; Boissy, J; Dourmad, J-Y

    2011-12-01

    The incorporation of feed-use (FU) amino acids (AAs) in diets results in a reduced use of protein-rich ingredients such as soybean meal, recognized to have elevated contributions to environmental impacts. This study investigated whether the incorporation of L-lysine.HCl, L-threonine and FU-methionine reduces the environmental impacts of pig and broiler feeds using Life Cycle Assessment. The following impact categories were considered: climate change, eutrophication, acidification, terrestrial ecotoxicity, cumulative energy demand and land occupation. Several feeds were formulated either to minimize the cost of the formulation (with or without AA utilization), to maximize AA incorporation (i.e. the cost of AA was considered to be similar to that of soybean meal), or to minimize greenhouse gas emissions. For both pig and broiler feeds, calculations were made first using only cereals and soybean meal as main ingredients and then using cereals and several protein-rich ingredients (soybean meal, rapeseed meal and peas). In addition, these calculations were performed using two types of soybean meal (from Brazil, associated with recent deforestation or not). For broiler feeds, two types of maize (from France, irrigated, with mineral fertilization v. not irrigated, with animal manure fertilization) were also tested. Regarding the feeds formulated to minimize cost, incorporation of AA decreased the values for eutrophication, terrestrial ecotoxicity and cumulative energy demand of both pig and broiler feeds, regardless of the base ingredients. Reduction in climate change and acidification due to the incorporation of AA depended on the nature of the feed ingredients, with the effect of AA incorporation being greater when combined with ingredients with high impacts such as soybean meal associated with deforestation. Feeds formulated to maximize AA incorporation generally had a similar composition to those formulated to minimize cost, suggesting that the costs of AA were not

  16. Improving thermal stability and light fastness of Acid Red 114 by incorporating its anions in a ZnAl-layered double hydroxides matrix

    Haifeng Zhu; Liren Wang; Pinggui Tang; Yongjun Feng; Dianqing Li

    2012-01-01

    Incorporation of anions of Acid Red 114 dye (1,3-naphthalenedisulfonic acid,8-[2-[3,3'-dimethyl-4'-[2-[4-[[(4-methylphenyl)sulfonyl]oxy] phenyl]diazenyl] [1,1'-biphenyl]-4-yl]diazenyl]-7-hydroxy-,disodium salt) (denoted as NPDA) into ZnAl-layered double hydroxides (LDHs) has been carried out by an anionexchange method in an effort to improve their thermal stability and light fastness.After intercalation of NPDA anions,the interlayer distance of the LDHs increases from 0.87 to 2.18nm,confirming their incorporation into the interlayer galleries of the LDHs host.Infrared spectroscopy and thermogravimetric analysis revealed the presence of host-guest interactions between LDHs layers and NPDA anions.The thermal stability of NPDA and ZnAl-NPDA-LDHs was compared by thermogravimetric-differential thermal analysis,UV-visible spectroscopy and infrared spectroscopy.It was found that the thermal stability of NPDA anions was markedly improved by incorporation into the ZnAl-LDHs matrix,while the light fastness was also enhanced.

  17. Thermal Behavior and Free-Radical-Scavenging Activity of Phytic Acid Alone and Incorporated in Cosmetic Emulsions

    André Luis Máximo Daneluti; Maria Valéria Robles Velasco; André Rolim Baby; Jivaldo do Rosário Matos

    2015-01-01

    Phytic acid is a natural compound widely used as depigmenting agent in cosmetic emulsions. Few studies are available in the literature covering the stability and the antioxidating property of this substance, used alone or into emulsions. Therefore, the purpose of this work was to investigate the thermal behavior and antioxidant properties of phytic acid alone and into cosmetic emulsions. The thermal behavior of this substance was evaluated by thermogravimetry (TG)/derivative thermogravimetr...

  18. Modified thermoresponsive Poloxamer 407 and chitosan sol-gels as potential sustained-release vaccine delivery systems

    Kojarunchitt, Thunjiradasiree; Baldursdottir, Stefania; Dong, Yao-Da;

    2015-01-01

    Thermoresponsive, particle-loaded, Poloxamer 407 (P407)-Pluronic-R® (25R4) or chitosan-methyl cellulose (MC) formulations were developed as single-dose, sustained release vaccines. The sol-gels, loaded either with a particulate vaccine (cubosomes) or soluble antigen (ovalbumin) and adjuvants (Quil...... antigen in vivo while the chitosan-MC sol-gels sustained the release of antigen up to at least 14 days after administration. The chitosan-MC sol-gels stimulated both cellular and humoral responses. The inclusion of cubosomes in the sol-gels did not provide a definitive beneficial effect. Further analysis...... of the formulations with small-angle X-ray scattering (SAXS) revealed that while cubosomes were stable in chitosan-MC gels they were not stable in P407-25R4 formulations. The reason for the mixed response to cubosome-loaded vehicles requires more investigation, however it appears that the cubosomes...

  19. Effects of two different dietary sources of long chain omega-3, highly unsaturated fatty acids on incorporation into the plasma, red blood cell, and skeletal muscle in horses.

    Hess, T M; Rexford, J K; Hansen, D K; Harris, M; Schauermann, N; Ross, T; Engle, T E; Allen, K G D; Mulligan, C M

    2012-09-01

    group compared with other groups and increased (P < 0.001) by 40% between d 30 and 90. As far as the authors are aware, this is the first study to demonstrate that dietary fatty acid supplementation will affect muscle fatty acid composition in horses. Incorporation of n-3 LCHUFA into blood and muscle depends directly on dietary supply of specific fatty acids. PMID:22966078

  20. Stable isotope labeling, in vivo, of D- and L-tryptophan pools in lemna gibba and the low incorporation of label into indole-3-acetic acid

    The authors present evidence that the role of tryptophan and other potential intermediates in the pathways that could lead to indole derivatives needs to be reexamined. Two lines of Lemna gibba were tested for uptake of [15N-indole]-labeled tryptophan isomers and incorporation of that label into free indole-3-acetic acid (IAA). Both lines required levels of L-[15N]tryptophan 2 to 3 orders of magnitude over endogenous levels in order to obtain measurable incorporation of label into IAA. Labeled L-tryptophan was extractable from plant tissue after feeding and showed no measurable isomerization into D-tryptophan. D-[15N]trytophan supplied to Lemna at rates of approximately 400 times excess of endogenous D-tryptophan levels (to yield an isotopic enrichment equal to that which allowed detection of the incorporation of L-tryptophan into IAA), did not result in measurable incorporation of label into free IAA. These results demonstrate that L-tryptophan is a more direct precursor to IAA than the D isomer and suggest (a) that the availability of tryptophan in vivo is not a limiting factor in the biosynthesis of IAA, thus implying that other regulatory mechanisms are in operation and (b) that L-tryptophan also may not be a primary precursor to IAA in plants

  1. Pharmacokinetics and enhanced oral bioavailability in beagle dogs of cyclosporine A encapsulated in glyceryl monooleate/poloxamer 407 cubic nanoparticles

    Jie Lai

    2009-12-01

    Full Text Available Jie Lai1,2, Yi Lu1, Zongning Yin2, Fuqiang Hu3, Wei Wu11School of Pharmacy, Fudan University, Shanghai, China, 2West China School of Pharmacy, Sichuan University, Chengdu, China, 3School of Pharmacy, Zhejiang University, Hangzhou, ChinaAbstract: Efforts to improve the oral bioavailability of cyclosporine A (CyA remains a challenge in the field of drug delivery. In this study, glyceryl monooleate (GMO/poloxamer 407 cubic nanoparticles were evaluated as potential vehicles to improve the oral bioavailability of CyA. Cubic nanoparticles were prepared via the fragmentation of a bulk GMO/poloxamer 407 cubic phase gel by sonication and homogenization. The cubic inner structure formed was verified using Cryo-TEM. The mean diameters of the nanoparticles were about 180 nm, and the entrapment efficiency of these particles for CyA was over 85%. The in vitro release of CyA from these nanoparticles was less than 5% at 12 h. The results of a pharmacokinetic study in beagle dogs showed improved absorption of CyA from cubic nanoparticles as compared to microemulsion-based Neoral®; higher Cmax (1371.18 ± 37.34 vs 969.68 ± 176.3 ng mL-1, higher AUC0–t (7757.21 ± 1093.64 vs 4739.52 ± 806.30 ng h mL-1 and AUC0–∞ (9004.77 ± 1090.38 vs 5462.31 ± 930.76 ng h mL-1. The relative oral bioavailability of CyA cubic nanoparticles calculated on the basis of AUC0–∞ was about 178% as compared to Neoral®. The enhanced bioavailability of CyA is likely due to facilitated absorption by cubic nanoparticles rather than improved release.Keywords: nanoparticles, cubosomes, cyclosporine A, glyceryl monooleate, oral drug delivery, bioavailability, beagle dogs

  2. Inter-tissue differences in fatty acid incorporation as a result of dietary oil manipulation in Port Jackson sharks (Heterodontus portusjacksoni).

    Beckmann, Crystal L; Mitchell, James G; Stone, David A J; Huveneers, Charlie

    2014-06-01

    Fatty acid profile analysis is a tool for dietary investigation that may complement traditional stomach contents analysis. While recent studies have shown that the liver of sharks fed different diets have differing fatty acid profiles, the degree to which diet is reflected in shark blood serum and muscle tissue is still poorly understood. An 18-week controlled feeding experiment was undertaken using captive Port Jackson sharks (Heterodontus portusjacksoni). Sharks were fed exclusive diets of artificial pellets treated with fish or poultry oil and sampled every 6 weeks. The fatty acid profiles from liver, blood serum, and muscle were affected differently, with the period from which significant differences were observed varying by tissue and diet type. The total fatty acid profiles of fish oil and poultry oil fed sharks were significantly different from week 12 onwards in the liver and blood serum, but significant differences were only observed by week 18 in the muscle tissue of sharks fed different diets. The drivers of dissimilarity which aligned with dietary input were 14:0, 18:2n-6, 20:5n-3, 18:1n-9 and 22:6n-3 in the liver and blood serum. Dietary fatty acids accumulated more consistently in the liver than in the blood plasma or muscle, likely due to its role as the central organ for fat processing and storage. Blood serum and muscle fatty acid profiles were influenced by diet, but fluctuated over-time. The low level of correlation between diet and muscle FA profiles is likely a result of low levels of fat (shark muscle tissues. Our findings describe inter-tissue differences in the incorporation of fatty acids from the diet to consumer, which should be taken into account when interpreting dietary patterns from fatty acid profiles. PMID:24719244

  3. TARTARIC ACID TEMPLATED SYNTHESIS OF MESOPOROUS Ti-INCORPORATED SILICA AND ITS CATALYTIC ACTIVITY FOR THE RING-OPENING POLYMERIZATION OF ε-CAPROLACTONE

    Jie-bin Pang; Chang-ming Dong; Kun-yuan Qiu; Yen Wei

    2002-01-01

    Ti-incorporated mesoporous silica materials with pore diameters of 3-4 nm have been prepared via the co-hydrolysis and co-condensation reactions, that is the sol-gel reactions, of titanium (Ⅳ) tetrabutoxide and tetraethylorthosilicate in the presence of tartaric acid as template, followed by extraction with ethanol to remove the templatemolecules. The materials were characterized in detail by Fourier transform infrared spectroscopy, nitrogen adsorption-desorption test, powder X-ray diffraction, transmission electron microscopy and X-ray energy dispersive spectroscopy. Theresults indicate that the Ti-containing silica materials have large specific surface areas (ca. 1200 m2 g-1) and pore volumes(ca. 0.900 cm3 g-1). The mesoporosity arises from disordered interconnecting channels or pores. The Ti-incorporated silicasexhibit catalytic activity for the ring-opening polymerization of ε-caprolactone, otherwise, the pure mesoporous silicamaterial shows no catalytic activity under the identical conditions.

  4. Peptide Synthesis through Cell-Free Expression of Fusion Proteins Incorporating Modified Amino Acids as Latent Cleavage Sites for Peptide Release.

    Liutkus, Mantas; Fraser, Samuel A; Caron, Karine; Stigers, Dannon J; Easton, Christopher J

    2016-05-17

    Chlorinated analogues of Leu and Ile are incorporated during cell-free expression of peptides fused to protein, by exploiting the promiscuity of the natural biosynthetic machinery. They then act as sites for clean and efficient release of the peptides simply by brief heat treatment. Dehydro analogues of Leu and Ile are similarly incorporated as latent sites for peptide release through treatment with iodine under cold conditions. These protocols complement enzyme-catalyzed methods and have been used to prepare calcitonin, gastrin-releasing peptide, cholecystokinin-7, and prolactin-releasing peptide prohormones, as well as analogues substituted with unusual amino acids, thus illustrating their practical utility as alternatives to more traditional chemical peptide synthesis. PMID:26918308

  5. CO2 incorporation and 4-hydroxy-2-methylbenzoic acid formation during anaerobic metabolism of m-cresol by a methanogenic consortium

    The metabolism of m-cresol by methanogenic cultures enriched from domestic sewage sludge was investigated. In the initial studies, bromoethanesulfonic acid was used to inhibit methane production. This led to the accumulation of 4.0 ± 0.8 mol of acetate per mol of m-cresol metabolized. These results suggested that CO2 incorporation occurred because each molecule of m-cresol contained seven carbon atoms, whereas four molecules of acetate product contained a total of eight carbon atoms. To verify this, [14C]bicarbonate was added to bromoethanesulfonic acid-inhibited cultures, and those cultures yielded [14C]acetate. Of the label recovered as acetate, 89% was found in the carboxyl position. Similar cultures fed [methyl-14C]m-cresol yielded methyl-labeled acetate. A 14C-labeled transient intermediate was detected in cultures given either m-cresol and [14C]bicarbonate or bicarbonate and [methyl-14C]m-cresol. The intermediate was identified as 4-hydroxy-2-methylbenzoic acid. In addition, another metabolite was detected and identified as 2-methylbenzoic acid. This compound appeared to be produced only sporadically, and it accumulated in the medium, suggested that the dehydroxylation of 4-hydroxy-2-methylbenzoic acid led to an apparent dead-end product

  6. Orchestrating the Biosynthesis of an Unnatural Pyrrolysine Amino Acid for Its Direct Incorporation into Proteins Inside Living Cells

    Ehrlich, M.; Gattner, M. J.; Viverge, B.; Bretzler, J.; Eisen, D.; Stadlmeier, M.; Vrábel, Milan; Carell, T.

    2015-01-01

    Roč. 21, č. 21 (2015), s. 7701-7704. ISSN 0947-6539 Institutional support: RVO:61388963 Keywords : amber suppression * bioorganic chemistry * pyrrolysine * synthetic biology * unnatural amino acid Subject RIV: CC - Organic Chemistry Impact factor: 5.731, year: 2014

  7. Incorporation of alpha-Ketoglutaric Acid as a Fixed Bed Scrubber Media for the Neutralization of Hydrazine Family Hypergolic Fuels

    DeVor, R. W.; Santiago-Maldonado, E.; Parkerson, J. K.

    2010-01-01

    A candidate scrubber media, alpha-ketoglutaric acid (aKGA) adsorbed onto a silica-based substrate was examined as a potential alternative to the hydrazine-family hypergolic fuel neutralization techniques currently utilized at NASA/Kennedy Space Center (KSC). Helvenson et. al. has indicated that aKGA will react with hydrazines to produce non-hazardous, possibly biodegradable products. Furthermore, the authors have previously tested and demonstrated the use of aKGA aqueous solutions as a replacement neutralizing agent for citric acid, which is currently used as a scrubbing agent in liquid scrubbers at KSC. Specific properties examined include reaction efficiency, the loading capacity of aKGA onto various silica substrates, and the comparison of aKGA media performance to that of the citric acid vapor scrubber systems at KSC and a commercial vapor scrubber media. Preliminary investigations showed hydrophobic aerogel particles to be an ideal substrate for the deposition of the aKGA. Current studies have shown that the laboratory produced aKGA-Aerogel absorbent media are more efficient and cost effective than a commercially available fixed bed scrubber media, although much less cost effective than liquid-based citric acid scrubbers (although possibly safer and less labor intensive). A comparison of all three alternative scrubber technologies (liquid aKGA, solid-phase aKGA, and commercially available sorbent materials) is given considering both hypergolic neutralization capabilities and relative costs (as compared to the current citric acid scrubbing technology in use at NASA/KSC).

  8. Fabrication of electrospun polylactic acid nanofilm incorporating cinnamon essential oil/β-cyclodextrin inclusion complex for antimicrobial packaging.

    Wen, Peng; Zhu, Ding-He; Feng, Kun; Liu, Fang-Jun; Lou, Wen-Yong; Li, Ning; Zong, Min-Hua; Wu, Hong

    2016-04-01

    A novel antimicrobial packaging material was obtained by incorporating cinnamon essential oil/β-cyclodextrin inclusion complex (CEO/β-CD-IC) into polylacticacid (PLA) nanofibers via electrospinning technique. The CEO/β-CD-IC was prepared by the co-precipitation method and SEM and FT-IR spectroscopy analysis indicated the successful formation of CEO/β-CD-IC, which improved the thermal stability of CEO. The CEO/β-CD-IC was then incorporated into PLA nanofibers by electrospinning and the resulting PLA/CEO/β-CD nanofilm showed better antimicrobial activity compared to PLA/CEO nanofilm. The minimum inhibitory concentration (MIC) of PLA/CEO/β-CD nanofilm against Escherichia coli and Staphylococcus aureus was approximately 1 mg/ml (corresponding CEO concentration 11.35 μg/ml) and minimum bactericidal concentration (MBC) was approximately 7 mg/ml (corresponding CEO concentration 79.45 μg/ml). Furthermore, compared with the casting method, the mild electrospinning process was more favorable for maintaining greater CEO in the obtained film. The PLA/CEO/β-CD nanofilm can effectively prolong the shelf life of pork, suggesting it has potential application in active food packaging. PMID:26593582

  9. Enhanced bone formation in electrospun poly(l-lactic-co-glycolic acid)-tussah silk fibroin ultrafine nanofiber scaffolds incorporated with graphene oxide.

    Shao, Weili; He, Jianxin; Sang, Feng; Wang, Qian; Chen, Li; Cui, Shizhong; Ding, Bin

    2016-05-01

    To engineer bone tissue, it is necessary to provide a biocompatible, mechanically robust scaffold. In this study, we fabricated an ultrafine nanofiber scaffold by electrospinning a blend of poly(l-lactic-co-glycolic acid), tussah silk fibroin, and graphene oxide (GO) and characterized its morphology, biocompatibility, mechanical properties, and biological activity. The data indicate that incorporation of 10wt.% tussah silk and 1wt.% graphene oxide into poly(l-lactic-co-glycolic acid) nanofibers significantly decreased the fiber diameter from 280 to 130nm. Furthermore, tussah silk and graphene oxide boosted the Young's modulus and tensile strength by nearly 4-fold and 3-fold, respectively, and significantly enhanced adhesion, proliferation in mouse mesenchymal stem cells and functionally promoted biomineralization-relevant alkaline phosphatase (ALP) and mineral deposition. The results indicate that composite nanofibers could be excellent and versatile scaffolds for bone tissue engineering. PMID:26952489

  10. Incorporation of [14C]-orotic acid into DNA of regenerating rat liver results in a greater specific radioactivity of DNA-5-methylcytosines than DNA-cytosines

    To examine in vivo organization of events related to 5-methylcytosine (5-MC) occurrence in DNA, they determined the distribution of orotic acid incorporation into 5-MC vs cytosine (C) of rat liver DNA during liver compensatory hyperplasia. Rats were injected with 5 μCi of [6-14C]-orotic acid at 2 h intervals from 12 to 26 h post-partial hepatectomy. After a 2-h incorporation, livers were removed, DNA was isolated by Marmur's procedure and hydrolyzed by formic acid. Individual pyrimidines were isolated, quantitated by preparative and analytical high pressure liquid chromatography, and specific radioactivities determined. Two peaks of DNA synthesis were observed: a major peak at 18 h and a smaller peak at 22-24 h. The specific radioactivity of labeled thymine in DNA was approximately twice that of labeled C from 16 to 24 h. A more complex pattern was seen for labeling in 5-MC vs C during the same period. Values for 5-MC/C ratios increased to a maximum of 1.5 from 16 to 20 h, then decreased to 1.0 from 20 to 26 h. Thus in regenerating rat liver, [14C]-orotic acid label occurred to a 1.5-fold greater extent among DNA-5-MC than DNA-C during DNA synthesis. These results suggest the existence of mechanisms for: (a) preferential DNA synthesis in regions enriched in 5-MC content, or (b) different rates of turnover for 5-MC and C during DNA replication

  11. Uptake and incorporation of labelled oleic acid and glycerol by isolated and perfused liver of Wistar rats

    After perfusion by oleic acid (9-103H) and glycerol (1-14C) previously starved Wistar rats, the synthesis of hepatic triglycerides (TG) and phospholipids (PL) follows the two following different method: during the first minutes of perfusions, the most important method synthesis of TG and especially of PL is a de novo synthesis utilizing glycerol and the exogenous fatty acids (FA). The TG synthesized are 18 : 1 18 : 1 18 : 1 and 16 : 0 18 : 1 18 : 2 the PL synthesized are LP, AP and LPC; during perfusions of long duration (30, 60, 120 min.), the major method of synthesis of TG and PL is an active exchange of FA of the endogenous glycerolipids. The TG synthesized are 16 : 0 18 : 1 18 : 1 and 16 : 0 18 : 1 18 : 2 the PL synthesized are PE and PC

  12. Enhanced Yield of Recombinant Proteins with Site-specifically Incorporated Unnatural Amino Acids Using a Cell-Free Expression System

    Smolskaya, Sviatlana; Zhang, Zhiwen Jonathan; Alfonta, Lital

    2013-01-01

    Using a commercial protein expression system, we sought the crucial elements and conditions for the expression of proteins with genetically encoded unnatural amino acids. By identifying the most important translational components, we were able to increase suppression efficiency to 55% and to increase mutant protein yields to levels higher than achieved with wild type expression (120%), reaching over 500 µg/mL of translated protein (comprising 25 µg in 50 µL of reaction mixture). To our knowle...

  13. Extending the cross-linking/mass spectrometry strategy: Facile incorporation of photo-activatable amino acids into the model protein calmodulin in Escherichia coli cells.

    Piotrowski, Christine; Ihling, Christian H; Sinz, Andrea

    2015-11-01

    Photo-induced cross-linking is a highly promising technique to investigate protein conformations and protein-protein interactions in their natural cellular environment. One strategy relies on the non-directed incorporation of diazirine-containing photo-activatable amino acids into proteins and a subsequent cross-link formation induced by UV-A irradiation. The advantage of this photo-cross-linking strategy is that it is not restricted to lysine residues and that hydrophobic regions in proteins can also be targeted, which is advantageous for investigating membrane proteins. Here, we present a simplified protocol that relies on the use of mineral salts medium without any special requirements for the incorporation of photo-methionines into proteins in Escherichia coli cells. The possibility to perform these experiments in E. coli is especially valuable as it is the major system for recombinant protein production. The method is exemplified for the Ca(2+) regulating protein calmodulin containing nine methionines, which were found to be replaced by their photo-activatable analogues. Our protocol allows the facile and stochastic incorporation of photo-methionines as the basis for conducting photo-cross-linking experiments in E. coli in an efficient manner. PMID:25726908

  14. Facile fabrication of poly(L-lactic acid) microsphere-incorporated calcium alginate/hydroxyapatite porous scaffolds based on Pickering emulsion templates.

    Hu, Yang; Ma, Shanshan; Yang, Zhuohong; Zhou, Wuyi; Du, Zhengshan; Huang, Jian; Yi, Huan; Wang, Chaoyang

    2016-04-01

    In this study, we develop a facile one-pot approach to the fabrication of poly(L-lactic acid) (PLLA) microsphere-incorporated calcium alginate (ALG-Ca)/hydroxyapatite (HAp) porous scaffolds based on HAp nanoparticle-stabilized oil-in-water Pickering emulsion templates, which contain alginate in the aqueous phase and PLLA in the oil phase. The emulsion aqueous phase is solidified by in situ gelation of alginate with Ca(2+) released from HAp by decreasing pH with slow hydrolysis of d-gluconic acid δ-lactone (GDL) to produce emulsion droplet-incorporated gels, followed by freeze-drying to form porous scaffolds containing microspheres. The pore structure of porous scaffolds can be adjusted by varying the HAp or GDL concentration. The compressive tests show that the increase of HAp or GDL concentration is beneficial to improve the compressive property of porous scaffolds, while the excessive HAp can lead to the decrease in compressive property. Moreover, the swelling behavior studies display that the swelling ratios of porous scaffolds reduce with increasing HAp or GDL concentration. Furthermore, hydrophobic drug ibuprofen (IBU) and hydrophilic drug bovine serum albumin (BSA) are loaded into the microspheres and scaffold matrix, respectively. In vitro drug release results indicate that BSA has a rapid release while IBU has a sustained release in the dual drug-loaded scaffolds. In vitro cell culture experiments verify that mouse bone mesenchymal stem cells can proliferate on the porous scaffolds well, indicating the good biocompatibility of porous scaffolds. All these results demonstrate that the PLLA microsphere-incorporated ALG-Ca/HAp porous scaffolds have a promising potential for tissue engineering and drug delivery applications. PMID:26774574

  15. Synergistic effect of pH-responsive folate-functionalized poloxamer 407-TPGS-mixed micelles on targeted delivery of anticancer drugs

    Butt AM; Mohd Amin MC; Katas H

    2015-01-01

    Adeel Masood Butt, Mohd Cairul Iqbal Mohd Amin, Haliza Katas Centre for Drug Delivery Research, Faculty of Pharmacy, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia Background: Doxorubicin (DOX), an anthracycline anticancer antibiotic, is used for treating various types of cancers. However, its use is associated with toxicity to normal cells and development of resistance due to overexpression of drug efflux pumps. Poloxamer 407 (P407) and vitamin E TPGS (d-α-tocopheryl poly...

  16. An attempt to stabilize tanshinone IIA solid dispersion by the use of ternary systems with nano-CaCO 3 and poloxamer 188

    Hong-mei Yan

    2014-01-01

    Full Text Available Background: Tanshinone IIA (TSIIA on solid dispersions (SDs has thermodynamical instability of amorphous drug. Ternary solid dispersions (tSDs can extend the stability of the amorphous form of drug. Poloxamer 188 was used as a SD carrier. Nano-CaCO 3 played an important role in adsorption of biomolecules and is being developed for a host of biotechnological applications. Objective: The aim of the present study was to investigate the dissolution behavior and accelerated stability of TSIIA on solid dispersions (SDs by the use of ternary systems with nano-CaCO 3 and poloxamer 188. Materials and Methods: The TSIIA tSDs were prepared by a spray-drying method. First, the effect of combination of poloxamer 188 and nano-CaCO 3 on TSIIA dissolution was studied. Subsequently, a set of complementary techniques (DSC, XRPD, SEM and FTIR was used to monitor the physical changes of TSIIA in the SDs. Finally, stability test was carried out under the conditions 40°C/75% RH for 6 months. Results: The characterization of tSDs by differential scanning calorimetry analysis (DSC and X-ray powder diffraction (XRPD showed that TSIIA was present in its amorphous form. Fourier transforms infrared spectroscopy (FTIR suggested the presence of interactions between TSIIA and carriers in tSDs. Improvement in the dissolution rate was observed for all SDs. The stability study conducted on SDs with nano-CaCO 3 showed stable drug content and dissolution behavior, over the period of 6 months as compared with freshly prepared SDs. Conclusion: SDs preparation with nano-CaCO 3 and poloxamer 188 may be a promising approach to enhance the dissolution and stability of TSIIA.

  17. Auxin-induced changes in the incorporation of /sup 3/H-amino acids into soybean ribosomal proteins

    Travis, R.L.; Key, J.L.

    1976-06-01

    Auxin-induced activation of 80S ribosomes and polyribosome formation in mature soybean (Glycine max var. Hawkeye) hypocotyl (R. L. Travis, J. M. Anderson, and J. L. Key. 1973. Plant Physiol. 52: 608-612) in the presence of a mixture of radioactive amino acids correlates with an increased specific radioactivity of at least three ribosomal proteins; the labeling of one of these increased severalfold above the control level. Results of experiments with 5-fluorouracil and cycloheximide indicated that the proteins in question were synthesized in response to auxin and became associated with pre-existing ribosomes. Ribosome dissociation experiments indicated that these proteins were associated with the 60S ribosome subunit.

  18. Synergistic effect of pH-responsive folate-functionalized poloxamer 407-TPGS-mixed micelles on targeted delivery of anticancer drugs

    Butt AM

    2015-02-01

    Full Text Available Adeel Masood Butt, Mohd Cairul Iqbal Mohd Amin, Haliza Katas Centre for Drug Delivery Research, Faculty of Pharmacy, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia Background: Doxorubicin (DOX, an anthracycline anticancer antibiotic, is used for treating various types of cancers. However, its use is associated with toxicity to normal cells and development of resistance due to overexpression of drug efflux pumps. Poloxamer 407 (P407 and vitamin E TPGS (d-α-tocopheryl polyethylene glycol succinate, TPGS are widely used polymers as drug delivery carriers and excipients for enhancing the drug retention times and stability. TPGS reduces multidrug resistance, induces apoptosis, and shows selective anticancer activity against tumor cells. Keeping in view the problems, we designed a mixed micelle system encapsulating DOX comprising TPGS for its selective anticancer activity and P407 conjugated with folic acid (FA for folate-mediated receptor targeting to cancer cells. Methods: FA-functionalized P407 was prepared by carbodiimide crosslinker chemistry. P407-TPGS/FA-P407-TPGS-mixed micelles were prepared by thin-film hydration method. Cytotoxicity of blank micelles, DOX, and DOX-loaded micelles was determined by alamarBlue® assay. Results: The size of micelles was less than 200 nm with encapsulation efficiency of 85% and 73% for P407-TPGS and FA-P407-TPGS micelles, respectively. Intracellular trafficking study using nile red-loaded micelles indicated improved drug uptake and perinuclear drug localization. The micelles show minimal toxicity to normal human cell line WRL-68, enhanced cellular uptake of DOX, reduced drug efflux, increased DOX–DNA binding in SKOV3 and DOX-resistant SKOV3 human ovarian carcinoma cell lines, and enhanced in vitro cytotoxicity as compared to free DOX. Conclusion: FA-P407-TPGS-DOX micelles show potential as a targeted nano-drug delivery system for DOX due to their multiple synergistic factors of selective anticancer

  19. Bacteriostatic and anti-collagenolytic dental materials through the incorporation of polyacrylic acid modified CuI nanoparticles

    Renne, Walter George; Mennito, Anthony Samuel; Schmidt, Michael Gerard; Vuthiganon, Jompobe; Chumanov, George

    2015-05-19

    Provided are antibacterial and antimicrobial surface coatings and dental materials by utilizing the antimicrobial properties of copper chalcogenide and/or copper halide (CuQ, where Q=chalcogens including oxygen, or halogens, or nothing). An antimicrobial barrier is created by incorporation of CuQ nanoparticles of an appropriate size and at a concentration necessary and sufficient to create a unique bioelectrical environment. The unique bioelectrical environment results in biocidal effectiveness through a multi-factorial mechanism comprising a combination of the intrinsic quantum flux of copper (Cu.sup.0, Cu.sup.1+, Cu.sup.2+) ions and the high surface-to-volume electron sink facilitated by the nanoparticle. The result is the constant quantum flux of copper which manifests and establishes the antimicrobial environment preventing or inhibiting the growth of bacteria. The presence of CuQ results in inhibiting or delaying bacterial destruction and endogenous enzymatic breakdown of the zone of resin inter-diffusion, the integrity of which is essential for dental restoration longevity.

  20. Mapping Substance P Binding Sites on the Neurokinin-1 Receptor Using Genetic Incorporation of a Photoreactive Amino Acid

    Valentin-Hansen, Louise; Park, Minyoung; Huber, Thomas;

    2014-01-01

    binding site for SP includes multiple domains in the N-terminal (Nt) segment and the second extracellular loop (ECLII) of NK1. To map precisely the NK1 residues that interact with SP, we applied a novel receptor-based targeted photocross-linking approach. We used amber codon suppression to introduce the...... photoreactive unnatural amino acid p-benzoyl-l-phenylalanine (BzF) at 11 selected individual positions in the Nt tail (residues 11-21) and 23 positions in the ECLII (residues 170(C-10)-193(C+13)) of NK1. The 34 NK1 variants were expressed in mammalian HEK293 cells and retained the ability to interact with a...

  1. A novel binuclear copper complex incorporating a nalidixic acid derivative displaying a one-dimensional coordination polymeric structure.

    Bergamini, F R G; Ribeiro, M A; Miranda, P C M L; Formiga, A L B; Corbi, P P

    2016-07-01

    The identification of the antibacterial action of nalidixic acid (nx) was central to the development of the quinolone antibacterial compounds. The ability of the nx naphthyridyl ring to interact with and inhibit some proteins has encouraged the investigation of similar structures in the search for more active compounds with less adverse effects. The possibility of structural modification by attachment of other biologically active moieties to the naphthyridyl ring of nx allowed the development of new active antimicrobial molecules. Hydrazone derivatives of nx can be synthesized easily based on the condensation of the hydrazide derivative of nx with the desired aldehyde or ketone. Only a few complexes with nx hydrazone derivatives have been described but for none were the crystal structures elucidated. The synthesis of a new one-dimensional Cu(II) coordination polymer, namely catena-poly[[copper(II)-di-μ-chlorido-copper(II)-{μ-1-ethyl-N'-[(1H-imidazol-4-yl)methylidene]-7-methyl-4-oxo-1,4-dihydro-1,8-naphthyridine-3-carbohydrazidato}-[dimethanolcopper(II)]-{μ-1-ethyl-N'-[(1H-imidazol-3-yl)methylidene]-7-methyl-4-oxo-1,4-dihydro-1,8-naphthyridine-3-carbohydrazidato}] dichloride methanol tetrasolvate], {[Cu3(C16H15N6O2)2Cl2(CH3OH)2]Cl2·4CH3OH}n, with the (1H-imidazol-4-yl)methylidene carbohydrazide derivative of nalidixic acid (denoted h4imi), is presented and its structure is compared to the density functional theory (DFT) optimized structure of free h4imi. The title structure presents an octahedral Cu(II) ion on an inversion centre alternating along a polymer chain with a square-pyramidal Cu(II) ion, with the two Cu(II) centres bridged by two chloride ligands. Hydrogen bonds involving chloride counter-ions and methanol solvent molecules mediate the three-dimensional packing of the polymer. Comparison of the geometrical results from the structure analysis with those derived from a DFT study of the free ligand reveal the differences that arise upon coordination

  2. In vitro characteristics of poly(lactic-co-glycolic acid) microspheres incorporating gelatin particles loading basic fibroblast growth factor

    Shao-hong LI; Shao-xi CAI; Bing LIU; Kai-wang MA; Zhen-ping WANG; Xiao-kun LI

    2006-01-01

    Aim: To construct a sustained drug release system for basic fibroblast growth factor (bFGF). With this special system, bFGF can be used to repair an injured peripheral nerve, injured spinal cord, or as a carrier for other drugs that need to be released over a long time. Methods: Microsphere composite was prepared by encapsulating bFGF into gelatin particles with poly(lactic-co-glycolic acid) (PLGA) as its outer-coating. The encapsulation was conducted by a phase separation method. Results: The average diameter of the gelatin particle-PLGA microsphere composite was 5-18 μm, and bFGF-loading efficiency was up to 80.5%. The bFGF releasing experiment indicated that this new composite system could release bFGF continuously and protect bFGF from denaturation. Conclusion: A modified approach was successfully employed to develop a biodegradable system for sustained release of the drug of bFGF in vitro.

  3. [Incorporation of labeled amino acids into proteins of organs and tissues of rabbit after cessation of vital activity of the body and during resuscitation].

    Konikova, A S; Pogosova, A V; Nikulin, V I

    1971-01-01

    Protein synthesis was assayed through incorporation of labelled amino acids (I-C14-glycine and S15-methionine) into proteins of organs and tissues of rabbit after cessation of life activity of the organism and during reanimation. It is established that after death, protein synthesis ceased in different organs and tissues not at the same time. First of all, protein synthesis stops in the spleen while in the pancreas this process persists for a longest time. Interruption of assimilatory processes after cessation of life is reversible, but restoration of the process of protein synthesis in various organs occurs not uniformly. Ambiguous restoration of precursor utilisation by organs in the process of protein biosynthesis reflects characteristics of the course of this assimilatory process in resuscitated+ animals at early stages after reanimation. PMID:5317617

  4. Enhancing microdialysis recovery of metal ions by incorporating poly-L-aspartic acid and poly-L-histidine in the perfusion liquid

    A study of the evaluation of poly-L-aspartic acid and poly-L-histidine as binding agents to enhance microdialysis recovery of metal ions is presented. Investigations were carried out to compare microdialysis recovery for Cr, Cu, Ni, and Pb when using water as the perfusion liquid as well as when using various concentrations of poly-L-aspartic acid and poly-L-histidine in the perfusion liquid. All experiments were carried out under quiescent conditions using a concentric type of microdialysis probe fitted with a polysulfone membrane having a 30 kDa molecular weight cut-off and a 10 mm effective dialysis length. The metal ions were determined using an electrothermal atomic absorption spectrometer equipped with a Zeemann background corrector. Incorporation of 0.032% (w/v) of poly-L-aspartic acid enhanced the recovery of Cu and Pb by factors of 90 and 64%, respectively (%RSD<3). The recovery of Cr was enhanced by 5%, but that of Ni never exceeded values achieved using ultra pure water. The use of 20% (w/v) of poly-L-histidine resulted in enhancement factors of 66 and 4% for Cu and Pb, respectively (%RSD<2). For both Cr and Ni, the recovery never exceeded that achieved with water. The data from these studies demonstrate the suitability of poly-L-aspartic and poly-L-histidine as selective and effective binding agents that enhance the microdialysis recovery of metal ions. Application of the optimised conditions to the determination of Pb and Cu in a wastewater sample confirmed the versatility of microdialysis, as higher recoveries of Cu were obtained with poly-L-aspartic acid compared to direct determination

  5. Selective incorporation of (15S)-hydroxyeicosatetraenoic acid in phosphatidylinositol of human neutrophils: Agonist-induced deacylation and transformation of stored hydroxyeicosanoids

    The uptake and mobilization of (15S)-hydroxy-5,8,11-cis-13-trans-eicosatetraenoic acid (15-HETE), a major product of arachidonic acid metabolism, was examined with human neutrophils. Upon exposure to labeled 15-HETE, PMNs rapidly (15 sec to 20 min) incorporated approximately 20% of the label into phosphatidylinositol, while less than 4% was associated with other phospholipid classes and neutral lipids. This pattern was distinct from that of either labeled arachidonate or labeled(5S)-hydroxy-8,11,14-cis-6-trans-eicosatetraenoic acid (5-HETE), which within 20 min were predominantly associated with triglycerides and phosphatidylcholine. After reversed-phase HPLC, greater than 98% of the label in phosphatidylinositol, isolated from PMNs, was released with phospholipase A2. Upon exposure to either chemotactic peptide (FMLP), phorbol 12-myristate 13-acetate, or an ionophore (A23187), 15-HETE-labeled PMNs released 15-HETE from phosphatidylinositol and displayed an impaired ability to generate leukotriene B4 (LTB4), 20-OH-LTB4, and 20-COOH-LTB4. Deacylated [3H]15-HETE was converted to (5S,15S)-dihydroxy-6,13-trans-8,11-cis-eicosatetraenoic acid (5,15-DHETE), lipoxin A4, and lipoxin B4, each carrying 3H label. PMNs labeled with 5-HETE also released and transformed this HETE when stimulated. However, the profile of labeled products differed between PMNs with either esterified 15-HETE or 5-HETE. When activated, 5-HETE-labeled PMNs generated both 5,20-DHETE and 5,15-DHETE but not labeled lipoxins. Threshold aggregation induced by FMLP with 15-HETE-labeled PMNs was inhibited, while the threshold response was relatively unimpaired with either A23187 or phorbol 12-myristate 13-acetate-induced aggregation. Results indicate that 15-HETE is esterified into phosphatidylinositol of PMNs, which can be mobilized and transformed upon exposure of the cells to a second signal

  6. Effect of acid or alkaline catalyst and of different capping agents on the optical properties of CdS nanoparticles incorporated within a diureasil hybrid matrix

    Gonçalves, Luis F. F. F.; Silva, Carlos J. R.; Kanodarwala, Fehmida K.; Stride, John A.; Pereira, Mario R.

    2015-11-01

    CdS nanoparticles (NPs) were synthesized using colloidal methods and incorporated within a diureasil hybrid matrix. The surface capping of the CdS NPs by 3-mercaptopropyltrimethoxysilane (MPTMS) and 3-aminopropyltrimethoxysilane (APTMS) organic ligands during the incorporation of the NPs within the hybrid matrix has been investigated. The matrix is based on poly(ethylene oxide)/poly(propylene oxide) chains grafted to a siliceous skeleton through urea bonds and was produced by sol-gel process. Both alkaline and acidic catalysis of the sol-gel reaction were used to evaluate the effect of each organic ligand on the optical properties of the CdS NPs. The hybrid materials were characterized by absorption, steady-state and time-resolved photoluminescence spectroscopy and High Resolution Transmission Electron Microscopy (HR-TEM). The preservation of the optical properties of the CdS NPs within the diureasil hybrids was dependent on the experimental conditions used. Both organic ligands (APTMS and MPTMS) demonstrated to be crucial in avoiding the increase of size distribution and clustering of the NPs within the hybrid matrix. The use of organic ligands was also shown to influence the level of interaction between the hybrid host and the CdS NPs. The CdS NPs showed large Stokes shifts and long average lifetimes, both in colloidal solution and in the xerogels, due to the origin of the PL emission in surface states. The CdS NPs capped with MPTMS have lower PL lifetimes compared to the other xerogel samples but still larger than the CdS NPs in the original colloidal solution. An increase in PL lifetimes of the NPs after their incorporation within the hybrid matrix is related to interaction between the NPs and the hybrid host matrix.

  7. Effect of feeding levels on the milk yield and on the incorporation of 14C labelled Volatile Fatty Acids in rumen into the milk

    It has been known that milk production was influenced by the feeding schedule, such as hay vs silage and ratio or concentrate to roughage, which have an important effect upon the VFA (volatile fatty acid) production in the rumen. This experiment were conducted using acetate-1-14C, propionate-1-14C, and butylate-1-14C, to examine the effect of nutritional feeding levels on the milk yield and on the incorporation rate of ruminal VFA-1-14C into the milk components. The experiment using six Holstein cows, had two periods, with different levels of concentrate feeding. In the first period, the concentrate fed was a half of 4%-FCM and in the second period, its level was a half of the first one. As a roughage, corn silage and grass hay were fed. Following the reduced feeding level of the concentrate, nutrient intake, milk yield, milk component yield and the lactose percentage were all lowered, but the fat percentage was slightly increased and the casein percentage was constant while the incorporation rates of each ruminal VFA-1-14C into the milk components were also lowered, remarkably for the lactose. (auth.)

  8. Hypolipidemic Effects of Aqueous Extract of Three Cultivars of Musa sapientum Fruit Peel on Poloxamer-407 Induced Hyperlipidemic Wistar Rats.

    C EDENTA

    2014-12-01

    Full Text Available Objective: The effect of aqueous extracts of the ripped fruit peel of three cultivars of Musa sapientum (Saro, Ominni and Oranta on the lipid profile of normolipidemic and hyperlipidemic rats were examined. Methods: Aqueous peel extracts of the 3-cultivars of Musa sapientum (100mg/Kg bw were administered to normolipidemic and poloxamer-407 induced hyperlidemic rats (140-180g. Atorvastatin was used as standard drug (70 mg/Kg bw. Blood samples were collected for determination of plasma total cholesterol (Tc, triacylglycerides (TAG, low-density lipoprotein cholesterol (LDL-c and high-density lipoprotein cholesterol (HDL-c after 21-days in normolipidemic rats and 3-days in hyperlipidemic rats. Results: Normolipidemic rats treated with extract of Saro showed a significant (p0.05 difference when compared to the Atorvastatin and normal control groups. Conclusion: The findings in this study reveals Oranta cultivar of M. sapientum as a better potential drug candidate among the cultivars studied, hence could be useful for the treatment of hyperlipidemia and other cardiovascular related diseases.

  9. Pharmacokinetic and milk penetration of a difloxacin long-acting poloxamer gel formulation with carboxy-methylcellulose in lactating goats.

    Escudero, Elisa; Marín, Pedro; Cárceles, Carlos M; Ramírez, María J; Fernández-Varón, Emilio

    2011-04-01

    The single-dose disposition kinetics of difloxacin were determined in clinically normal lactating goats (n=6) after subcutaneous administration of a long-acting poloxamer 407 gel formulation with carboxy-methylcellulose (P407-CMC). Difloxacin concentrations were determined by high performance liquid chromatography with fluorescence detection. The concentration-time data were analysed by non-compartmental kinetic methods. Plasma and milk elimination half-lives after P407-CMC dosing were 35.19 h and 33.93 h, respectively. With this formulation, difloxacin achieved maximum plasma concentrations of 2.67±0.34 mg/L at 2.92±1.20 h and maximum milk concentrations of 2.31±0.35 mg/L at 4.00±0.00 h. The area under the curve (AUC) ratio AUC(milk)/AUC(plasma) was 0.89 after P407-CMC administration. It was concluded that a 15 mg/kg dose of difloxacin within P407-CMC would be effective against mastitis pathogens with a minimum inhibitory concentration (MIC)≤0.12 mg/L. PMID:20359917

  10. Dual-functional transdermal drug delivery system with controllable drug loading based on thermosensitive poloxamer hydrogel for atopic dermatitis treatment

    Wang, Wenyi; Wat, Elaine; Hui, Patrick C. L.; Chan, Ben; Ng, Frency S. F.; Kan, Chi-Wai; Wang, Xiaowen; Hu, Huawen; Wong, Eric C. W.; Lau, Clara B. S.; Leung, Ping-Chung

    2016-04-01

    The treatment of atopic dermatitis (AD) has long been viewed as a problematic issue by the medical profession. Although a wide variety of complementary therapies have been introduced, they fail to combine the skin moisturizing and drug supply for AD patients. This study reports the development of a thermo-sensitive Poloxamer 407/Carboxymethyl cellulose sodium (P407/CMCs) composite hydrogel formulation with twin functions of moisture and drug supply for AD treatment. It was found that the presence of CMCs can appreciably improve the physical properties of P407 hydrogel, which makes it more suitable for tailored drug loading. The fabricated P407/CMCs composite hydrogel was also characterized in terms of surface morphology by field emission scanning electron microscopy (FE-SEM), rheological properties by a rheometer, release profile in vitro by dialysis method and cytotoxicity test. More importantly, the findings from transdermal drug delivery behavior revealed that P407/CMCs showed desirable percutaneous performance. Additionally, analysis of cytotoxicity test suggested that P407/CMCs composite hydrogel is a high-security therapy for clinical trials and thus exhibits a promising way to treat AD with skin moisturizing and medication.

  11. Self-assembled silk sericin/poloxamer nanoparticles as nanocarriers of hydrophobic and hydrophilic drugs for targeted delivery

    In recent times self-assembled micellar nanoparticles have been successfully employed in tissue engineering for targeted drug delivery applications. In this review, silk sericin protein from non-mulberry Antheraea mylitta tropical tasar silk cocoons was blended with pluronic F-127 and F-87 in the presence of solvents to achieve self-assembled micellar nanostructures capable of carrying both hydrophilic (FITC-inulin) and hydrophobic (anticancer drug paclitaxel) drugs. The fabricated nanoparticles were subsequently characterized for their size distribution, drug loading capability, cellular uptake and cytotoxicity. Nanoparticle sizes ranged between 100 and 110 nm in diameter as confirmed by dynamic light scattering. Rapid uptake of these particles into cells was observed in in vitro cellular uptake studies using breast cancer MCF-7 cells. In vitro cytotoxicity assay using paclitaxel-loaded nanoparticles against breast cancer cells showed promising results comparable to free paclitaxel drugs. Drug-encapsulated nanoparticle-induced apoptosis in MCF-7 cells was confirmed by FACS and confocal microscopic studies using Annexin V staining. Up-regulation of pro-apoptotic protein Bax, down-regulation of anti-apoptotic protein Bcl-2 and cleavage of regulatory protein PARP through Western blot analysis suggested further drug-induced apoptosis in cells. This study projects silk sericin protein as an alternative natural biomaterial for fabrication of self-assembled nanoparticles in the presence of poloxamer for successful delivery of both hydrophobic and hydrophilic drugs to target sites.

  12. Self-assembled silk sericin/poloxamer nanoparticles as nanocarriers of hydrophobic and hydrophilic drugs for targeted delivery

    Mandal, Biman B; Kundu, S C, E-mail: kundu@hijli.iitkgp.ernet.i [Department of Biotechnology, Indian Institute of Technology, Kharagpur 721302 (India)

    2009-09-02

    In recent times self-assembled micellar nanoparticles have been successfully employed in tissue engineering for targeted drug delivery applications. In this review, silk sericin protein from non-mulberry Antheraea mylitta tropical tasar silk cocoons was blended with pluronic F-127 and F-87 in the presence of solvents to achieve self-assembled micellar nanostructures capable of carrying both hydrophilic (FITC-inulin) and hydrophobic (anticancer drug paclitaxel) drugs. The fabricated nanoparticles were subsequently characterized for their size distribution, drug loading capability, cellular uptake and cytotoxicity. Nanoparticle sizes ranged between 100 and 110 nm in diameter as confirmed by dynamic light scattering. Rapid uptake of these particles into cells was observed in in vitro cellular uptake studies using breast cancer MCF-7 cells. In vitro cytotoxicity assay using paclitaxel-loaded nanoparticles against breast cancer cells showed promising results comparable to free paclitaxel drugs. Drug-encapsulated nanoparticle-induced apoptosis in MCF-7 cells was confirmed by FACS and confocal microscopic studies using Annexin V staining. Up-regulation of pro-apoptotic protein Bax, down-regulation of anti-apoptotic protein Bcl-2 and cleavage of regulatory protein PARP through Western blot analysis suggested further drug-induced apoptosis in cells. This study projects silk sericin protein as an alternative natural biomaterial for fabrication of self-assembled nanoparticles in the presence of poloxamer for successful delivery of both hydrophobic and hydrophilic drugs to target sites.

  13. Self-assembled silk sericin/poloxamer nanoparticles as nanocarriers of hydrophobic and hydrophilic drugs for targeted delivery

    Mandal, Biman B.; Kundu, S. C.

    2009-09-01

    In recent times self-assembled micellar nanoparticles have been successfully employed in tissue engineering for targeted drug delivery applications. In this review, silk sericin protein from non-mulberry Antheraea mylitta tropical tasar silk cocoons was blended with pluronic F-127 and F-87 in the presence of solvents to achieve self-assembled micellar nanostructures capable of carrying both hydrophilic (FITC-inulin) and hydrophobic (anticancer drug paclitaxel) drugs. The fabricated nanoparticles were subsequently characterized for their size distribution, drug loading capability, cellular uptake and cytotoxicity. Nanoparticle sizes ranged between 100 and 110 nm in diameter as confirmed by dynamic light scattering. Rapid uptake of these particles into cells was observed in in vitro cellular uptake studies using breast cancer MCF-7 cells. In vitro cytotoxicity assay using paclitaxel-loaded nanoparticles against breast cancer cells showed promising results comparable to free paclitaxel drugs. Drug-encapsulated nanoparticle-induced apoptosis in MCF-7 cells was confirmed by FACS and confocal microscopic studies using Annexin V staining. Up-regulation of pro-apoptotic protein Bax, down-regulation of anti-apoptotic protein Bcl-2 and cleavage of regulatory protein PARP through Western blot analysis suggested further drug-induced apoptosis in cells. This study projects silk sericin protein as an alternative natural biomaterial for fabrication of self-assembled nanoparticles in the presence of poloxamer for successful delivery of both hydrophobic and hydrophilic drugs to target sites.

  14. SBA-15-incorporated nanoscale zero-valent iron particles for chromium(VI) removal from groundwater: Mechanism, effect of pH, humic acid and sustained reactivity

    Sun, Xia, E-mail: lygsunxia@163.com [School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094 (China); School of Chemistry and Chemical Engineering, Huaihai Institute of Technology, Lianyungang 222005 (China); Yan, Yubo [School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094 (China); Li, Jiansheng, E-mail: lijsh@mail.njust.edu.cn [School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094 (China); Han, Weiqing [School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094 (China); Wang, Lianjun, E-mail: wanglj@mail.njust.edu.cn [School of Environmental and Biological Engineering, Nanjing University of Science and Technology, Nanjing 210094 (China)

    2014-02-15

    Highlights: • Zero-valent iron nanoparticles were incorporated in the pores of SBA-15 rods. • Mechanism of the Cr(VI) removal by NZVIs/SBA-15 was proposed. • A low pH value was in favor of the Cr(VI) removal. • Humic acid (HA) had a negligible effect on the reactivity of NZVIs/SBA-15. • The stable reduction of NZVIs/SBA-15 was observed within six cycles. -- Abstract: Nanoscale zero-valent iron particles (NZVIs) were incorporated inside the channels of SBA-15 rods by a “two solvents” reduction technique and used to remove Cr(VI) from groundwater. The resulting NZVIs/SBA-15 composites before and after reaction were characterized by N{sub 2} adsorption/desorption, X-ray diffraction (XRD), transmission electron microscopy (TEM) and X-ray photoelectron spectroscopy (XPS). Results helped to propose the mechanism of Cr(VI) removal by NZVIs/SBA-15, where Cr(VI) in aqueous was firstly impregnated into the channels of the silica, then adsorbed on the surfaces of the incorporated NZVIs and reduced to Cr(III) directly in the inner pores of the silica. Corrosion products included Fe{sub 2}O{sub 3}, FeO(OH), Fe{sub 3}O{sub 4} and Cr{sub 2}FeO{sub 4}. Batch experiments revealed that Cr(VI) removal decreased from 99.7% to 92.8% when the initial solution pH increased from 5.5 to 9.0, accompanied by the decrease of the k{sub obs} from 0.600 to 0.024 min{sup −1}. Humic acid (HA) had a little effect on the removal efficiency of Cr(VI) by NZVIs/SBA-15 but could decrease the reduction rate. The stable reduction of NZVIs/SBA-15 was observed within six cycles. NZVIs/SBA-15 composites offer a promising alternative material to remove heavy metals from groundwater.

  15. Chronic Dosing with Membrane Sealant Poloxamer 188 NF Improves Respiratory Dysfunction in Dystrophic Mdx and Mdx/Utrophin-/- Mice.

    Bruce E Markham

    Full Text Available Poloxamer 188 NF (national formulary (NF grade of P-188 improves cardiac muscle function in the mdx mouse and golden retriever muscular dystrophy models. However in vivo effects on skeletal muscle have not been reported. We postulated that P-188 NF might protect diaphragm muscle membranes from contraction-induced injury in mdx and mdx/utrophin-/- (dko muscular dystrophy models. In the first study 7-month old mdx mice were treated for 22 weeks with subcutaneous (s.c. injections of saline or P-188 NF at 3 mg/Kg. In the second, dkos were treated with saline or P-188 NF (1 mg/Kg for 8 weeks beginning at age 3 weeks. Prednisone was the positive control in both studies. Respiratory function was monitored using unrestrained whole body plethysmography. P-188 NF treatment affected several respiratory parameters including tidal volume/BW and minute volume/BW in mdx mice. In the more severe dko model, P-188 NF (1 mg/Kg significantly slowed the decline in multiple respiratory parameters compared with saline-treated dko mice. Prednisone's effects were similar to those seen with P-188 NF. Diaphragms from P-188 NF or prednisone treated mdx and dko mice showed signs of muscle fiber protection including less centralized nuclei, less variation in fiber size, greater fiber density, and exhibited a decreased amount of collagen deposition. P-188 NF at 3 mg/Kg s.c. also improved parameters of systolic and diastolic function in mdx mouse hearts. These results suggest that P-188 NF may be useful in treating respiratory and cardiac dysfunction, the leading causes of death in Duchenne muscular dystrophy patients.

  16. Preparation and characterization of celecoxib solid dispersions; comparison of poloxamer-188 and PVP-K30 as carriers

    Alireza Homayouni

    2014-05-01

    Full Text Available Objective(s:Solid dispersion formulation is the most promising strategy to improve oral bioavailability of poorly water soluble drugs. The aim of this study was to compare the effect of polyvinylpyrrolidone K30 (PVP and poloxamer-188 (PLX as carrier in solid dispersion formulations of celecoxib (CLX. Materials and Methods: Solid dispersions of CLX:PVP or CLX:PLX were prepared at different ratios (2:1, 1:1, 1:2, 1:4, 1:6 by solvent evaporation and melting methods, respectively. The characterization of samples was performed using differential scanning calorimetery (DSC, X-Ray powder diffraction (XRPD and Fourier transform infrared spectroscopy (FT-IR. The Gordon-Taylor equation was used to estimate the Tg of solid dispersion systems and the possibility of the interaction between CLX and PVP. Also, the dissolution rate of all samples was determined. Results: DSC and XRPD analyses confirmed the presence of amorphous state of drug in solid dispersion systems. FT-IR studies showed CLX could participate in hydrogen bonding with PVP whilst no specific interaction between CLX and PLX was observed. Both PVP and PLX enhanced the dissolution rate of drug in solid dispersion samples. The dissolution rate was dependent on the ratio of drug: carrier. Interestingly, the solid dispersion samples of PLX at 2:1 and 1:1 drug: carrier showed slower dissolution rate than pure CLX, whilst these results were not observed for PVP. Conclusion: The effect of PVP on dissolution rate enhancement was more pronounced compared to the other carrier. Having a higher Tg and more effect on dissolution rate, PVP could be considered as a more suitable carrier compared to PLX in solid dispersion formulation of CLX.

  17. iHyd-PseAAC: Predicting Hydroxyproline and Hydroxylysine in Proteins by Incorporating Dipeptide Position-Specific Propensity into Pseudo Amino Acid Composition

    Yan Xu

    2014-05-01

    Full Text Available Post-translational modifications (PTMs play crucial roles in various cell functions and biological processes. Protein hydroxylation is one type of PTM that usually occurs at the sites of proline and lysine. Given an uncharacterized protein sequence, which site of its Pro (or Lys can be hydroxylated and which site cannot? This is a challenging problem, not only for in-depth understanding of the hydroxylation mechanism, but also for drug development, because protein hydroxylation is closely relevant to major diseases, such as stomach and lung cancers. With the avalanche of protein sequences generated in the post-genomic age, it is highly desired to develop computational methods to address this problem. In view of this, a new predictor called “iHyd-PseAAC” (identify hydroxylation by pseudo amino acid composition was proposed by incorporating the dipeptide position-specific propensity into the general form of pseudo amino acid composition. It was demonstrated by rigorous cross-validation tests on stringent benchmark datasets that the new predictor is quite promising and may become a useful high throughput tool in this area. A user-friendly web-server for iHyd-PseAAC is accessible at http://app.aporc.org/iHyd-PseAAC/. Furthermore, for the convenience of the majority of experimental scientists, a step-by-step guide on how to use the web-server is given. Users can easily obtain their desired results by following these steps without the need of understanding the complicated mathematical equations presented in this paper just for its integrity.

  18. Incorporation of nisI-mediated nisin immunity improves vector-based nisin-controlled gene expression in lactic acid bacteria.

    Oddone, Gian M; Mills, David A; Block, David E

    2009-05-01

    Lactic acid bacteria (LAB) have been used successfully to express a wide variety of recombinant proteins, ranging from flavor-active proteins to antibiotic peptides and oral vaccines. The nisin-controlled expression (NICE) system is the most prevalent of the systems for production of heterologous proteins in LAB. Previous optimization of the NICE system has revealed a strong limit on the concentration of the inducer nisin that can be tolerated by the culture of host cells. In this work, the nisin immunity gene, nisI, has been inserted into the recently reported pMSP3535H2 vector that contains the complete NICE system on a high-copy Escherichia coli-LAB shuttle vector. Fed-batch fermentation data show that Lactococcus lactis IL1403 cells transformed with the new vector, pMSP3535H3, tolerate a 5-fold increase in the concentration of the inducer nisin, and, at this elevated concentration, produce a 1.8-fold increased level of green fluorescent protein (GFP), a model recombinant protein. Therefore, the incorporation of nisI in the pMSP3535H3 NICE system described here unveils new ranges of induction parameters to be studied in the course of optimizing recombinant protein expression in LAB. PMID:19141301

  19. GOASVM: a subcellular location predictor by incorporating term-frequency gene ontology into the general form of Chou's pseudo-amino acid composition.

    Wan, Shibiao; Mak, Man-Wai; Kung, Sun-Yuan

    2013-04-21

    Prediction of protein subcellular localization is an important yet challenging problem. Recently, several computational methods based on Gene Ontology (GO) have been proposed to tackle this problem and have demonstrated superiority over methods based on other features. Existing GO-based methods, however, do not fully use the GO information. This paper proposes an efficient GO method called GOASVM that exploits the information from the GO term frequencies and distant homologs to represent a protein in the general form of Chou's pseudo-amino acid composition. The method first selects a subset of relevant GO terms to form a GO vector space. Then for each protein, the method uses the accession number (AC) of the protein or the ACs of its homologs to find the number of occurrences of the selected GO terms in the Gene Ontology annotation (GOA) database as a means to construct GO vectors for support vector machines (SVMs) classification. With the advantages of GO term frequencies and a new strategy to incorporate useful homologous information, GOASVM can achieve a prediction accuracy of 72.2% on a new independent test set comprising novel proteins that were added to Swiss-Prot six years later than the creation date of the training set. GOASVM and Supplementary materials are available online at http://bioinfo.eie.polyu.edu.hk/mGoaSvmServer/GOASVM.html. PMID:23376577

  20. Poloxamer bioadhesive hydrogel for buccal drug delivery: Cytotoxicity and trans-epithelial permeability evaluations using TR146 human buccal epithelial cell line.

    Zeng, Ni; Mignet, Nathalie; Dumortier, Gilles; Olivier, Elodie; Seguin, Johanne; Maury, Marc; Scherman, Daniel; Rat, Patrice; Boudy, Vincent

    2015-11-30

    A salbutamol sulfate (SS)-Poloxamer bioadhesive hydrogel specially developed for buccal administration was investigated by studying interactions with TR146 human buccal epithelium cells (i.e. cellular toxicity (i) and trans-epithelial SS diffusion (ii)). The assessment of cell viability (MTT, Alamar Blue), membrane integrity (Neutral Red), and apoptosis assay (Hoechst 33342), were performed and associated to Digital Holographic Microscopy analysis. After the treatment of 2h, SS solution induced drastic cellular alterations that were prevented by hydrogels in relation with the concentrations of poloxamer and xanthan gum. The formulation containing P407 19%/P188 1%/Satiaxane 0.1% showed the best tolerance after single and multiple administrations and significantly reduced the trans-epithelial permeability from 5.00±0.29 (×10(3)) (SS solution) to 1.83±0.22 cm/h. Digital Holographic Microscopy images in good agreement with the viability data confirmed the great interest of this direct technique. In conclusion, the proposed hydrogels represent a safe and efficient buccal drug delivery platform. PMID:26403384

  1. Rhodanese incorporated in Langmuir and Langmuir-Blodgett films of dimyristoylphosphatidic acid: Physical chemical properties and improvement of the enzyme activity.

    de Araújo, Felipe Tejada; Caseli, Luciano

    2016-05-01

    Preserving the catalytic activity of enzymes immobilized in bioelectronics devices is essential for optimal performance in biosensors. Therefore, ultrathin films in which the architecture can be controlled at the molecular level are of interest. In this work, the enzyme rhodanese was adsorbed onto Langmuir monolayers of the phospholipid dimyristoylphosphatidic acid and characterized by surface pressure-area isotherms, polarization-modulated infrared reflection-absorption spectroscopy (PM-IRRAS), and Brewster angle microscopy (BAM). The incorporation of the enzyme (5% in mol) in the lipid monolayer expanded the film, providing small surface domains, as visualized by BAM. Also, amide bands could be identified in the PM-IRRAS spectra, confirming the presence of the enzyme at the air-water interface. Structuring of the enzyme into α-helices was identified in the mixed monolayer and was preserved when the film was transferred from the liquid interface to solids supports as Langmuir-Blodgett (LB) films. The enzyme-lipid LB films were then characterized by fluorescence spectroscopy, PM-IRRAS, and atomic force microscopy. Measurements of the catalytic activity towards cyanide showed that the enzyme accommodated in the LB films preserved more than 87% of the enzyme activity in relation to the homogeneous medium. After 1 month, the enzyme in the LB film maintained 85% of the activity in contrast to the homogeneous medium, which 24% of the enzyme activity was kept. The method presented in this work not only points to an enhanced catalytic activity toward cyanide, but also may explain why certain film architectures exhibit an improved performance. PMID:26836478

  2. Incorporation of EPA and DHA into plasma phospholipids in response to different omega-3 fatty acid formulations - a comparative bioavailability study of fish oil vs. krill oil

    Schuchardt Jan

    2011-08-01

    Full Text Available Abstract Background Bioavailability of omega-3 fatty acids (FA depends on their chemical form. Superior bioavailability has been suggested for phospholipid (PL bound omega-3 FA in krill oil, but identical doses of different chemical forms have not been compared. Methods In a double-blinded crossover trial, we compared the uptake of three EPA+DHA formulations derived from fish oil (re-esterified triacylglycerides [rTAG], ethyl-esters [EE] and krill oil (mainly PL. Changes of the FA compositions in plasma PL were used as a proxy for bioavailability. Twelve healthy young men (mean age 31 y were randomized to 1680 mg EPA+DHA given either as rTAG, EE or krill oil. FA levels in plasma PL were analyzed pre-dose and 2, 4, 6, 8, 24, 48, and 72 h after capsule ingestion. Additionally, the proportion of free EPA and DHA in the applied supplements was analyzed. Results The highest incorporation of EPA+DHA into plasma PL was provoked by krill oil (mean AUC0-72 h: 80.03 ± 34.71%*h, followed by fish oil rTAG (mean AUC0-72 h: 59.78 ± 36.75%*h and EE (mean AUC0-72 h: 47.53 ± 38.42%*h. Due to high standard deviation values, there were no significant differences for DHA and the sum of EPA+DHA levels between the three treatments. However, a trend (p = 0.057 was observed for the differences in EPA bioavailability. Statistical pair-wise group comparison's revealed a trend (p = 0.086 between rTAG and krill oil. FA analysis of the supplements showed that the krill oil sample contained 22% of the total EPA amount as free EPA and 21% of the total DHA amount as free DHA, while the two fish oil samples did not contain any free FA. Conclusion Further studies with a larger sample size carried out over a longer period are needed to substantiate our findings and to determine differences in EPA+DHA bioavailability between three common chemical forms of LC n-3 FA (rTAG, EE and krill oil. The unexpected high content of free EPA and DHA in krill oil, which might have a

  3. Incorporation of nano-clay saponite layers in the organo-clay hybrid films using anionic amphiphile stearic acid by Langmuir–Blodgett technique

    Hussain, Syed Arshad, E-mail: sa_h153@hotmail.com [Department of Physics, Tripura University, Suryamaninagar-799022 (India); Chakraborty, S.; Bhattacharjee, D. [Department of Physics, Tripura University, Suryamaninagar-799022 (India); Schoonheydt, R.A. [Centres for Surface Chemistry and Catalysis, K.U. Leuven, Kasteelpark Arenberg 23, 3001 Leuven (Belgium)

    2013-06-01

    In general cationic amphiphiles are used to prepare organo-clay hybrid film in Langmuir–Blodgett (LB) technique. In this present communication we demonstrated a unique technique to prepare the organo–clay hybrid films using an anionic amphiphile. The T–O–T type clay saponite was incorporated onto a floating stearic acid monolayer via a divalent cation Mg{sup 2+}. Salt MgCl{sub 2} was mixed along with the clay dispersion in the LB trough and amphiphile solution was spread onto the subphase in order to make the organo-clay hybrid films. It was observed that salt (MgCl{sub 2}) concentration on the subphase affects the organization of nano-dimensional clay platelet (saponite) in organo-clay hybrid films at air–water interface as well as in LB films. Noticeable changes in area per molecule and shape of the isotherms were observed and measured at subphases with different salt concentrations. Infrared reflection absorption spectroscopy studies reveal that only an in-plane (996 cm{sup −1}) vibration of ν (Si-O) band occurred when the salt concentration was 10 mM. However, both in-plane (996 cm{sup −1}) and out-of-plane (1063 cm{sup −1}) vibrations of the ν (Si-O) band of saponite occurred when the subphase salt concentration was 100 mM. Also the out-of-plane vibration of ν (OH) of saponite was prominent at higher salt concentration. This is because at lower salt concentration clay sheets remain flat on the surface whereas; at higher MgCl{sub 2} concentration they aggregated and form stacks of saponite layers. Also they may be slightly tilted with a very small tilt angle at higher salt concentration making a favorable condition for both in-plane and out-of-plane vibrations of ν (Si-O) in the hybrid films. Observed decrease in starting area per molecule in the pressure area isotherm measured at higher salt concentration also supports the tilting of clay layers at air–clay dispersion interface. Attentuated total reflectance Fourier transform infrared

  4. An Integrated Sequence-Structure Database incorporating matching mRNA sequence, amino acid sequence and protein three-dimensional structure data.

    Adzhubei, I A; Adzhubei, A. A.; Neidle, S.

    1998-01-01

    We have constructed a non-homologous database, termed the Integrated Sequence-Structure Database (ISSD) which comprises the coding sequences of genes, amino acid sequences of the corresponding proteins, their secondary structure and straight phi,psi angles assignments, and polypeptide backbone coordinates. Each protein entry in the database holds the alignment of nucleotide sequence, amino acid sequence and the PDB three-dimensional structure data. The nucleotide and amino acid sequences for ...

  5. Effects of 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin on (3H)-thymidine incorporation into rat liver deoxyribonucleic acid

    The effect of 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin (TCDD) on (3H)-thymidine incorporation into hepatic DNA was studied in rats. In nonhepatectomized male and female animals, incorporation measured at the peak of the first round of liver DNA synthesis after TCDD treatment (10 ug/kg) was similar to that of control animals. In contrast, the first round of (3H) thymidine incorporation after a 1/3 hepatectomy was enhanced 3-fold in TCDD-treated rats. The enhanced response to 1/3 hepatectomy was produced by doses of TCDD ranging from 1 to 30 ug/kg with an apparent ED50 of 5 ug/kg. Enhanced incorporation was observed when the 1/3 hepatectomy was performed 5-10 days after an ED50 dose and it returned to the control level after 20 days. This enhanced response was not preceded by changes in food consumption or hepatic activities of ornithine decarboxylase (ODC), tyrosine aminotransferase (ITAT) or gamma glutamyl transpeptidase (GGT) when compared to respective control values. Also, the enhanced incorporation was not necessarily due to removal of 1/3 of the liver because it was also seen in TCDD-treated rats that were laparotomized. The mechanism of enhancement in laparatomized animals does not appear to involve a diminished response of the liver to the inhibitory effects of adrenal hormones on liver DNA synthesis. This was suggested by the finding that an adrenalectomy prior to the laparotomy did not block the enhanced incorporation of (3H) thymidine into hepatic DNA. The mechanism by which TCDD enhances the first round of liver DNA synthesis after a 1/3 hepatectomy or laparotomy remains to be determined. (author)

  6. Incorporation of poly(glycidylmethacrylate) grafted bacterial cellulose nano-whiskers in poly(lactic acid) nanocomposites: improved barrier and mechanical properties

    Poly(glycidyl methacrylate) (PGMA) was grafted onto bacterial cellulose nanowhiskers (BCNW) by means of a redox-initiated free radical copolymerization reaction. The incorporation of PGMA chains decreased the thermal stability and crystallinity of BCNW. The neat and the PGMA-grafted BCNW were subseq...

  7. Task-specific ionic liquids incorporating alkyl phosphate cations for extraction of U(VI) from nitric acid medium. Synthesis, characterization, and extraction performance

    Imidazole-based task-specific ionic liquids designed to extract U(VI) from nitric acid medium were synthesized through the introduction of tributyl phosphate functional group in cationic of these ionic liquids. Their structures were confirmed by Fourier transform infrared spectrometer, nuclear magnetic resonance spectrum and element analysis. These task-specific ionic liquids can be used simultaneously as solvents and extractants for extracting U(VI) in nitric acid medium. Preliminary extraction data showed that they could be efficiently used to extract U(VI) in nitric acid medium at room temperature. (author)

  8. Optical, morphology and electrical properties of In2O3 incorporating acid-treated single-walled carbon nanotubes based DSSC

    This study focuses on the influence of an acid treatment process of single-walled carbon nanotubes (SWCNTs) in In2O3-based dye-sensitized solar cells (DSSCs). Pure In2O3, In2O3-SWCNTs with acid treatment and In2O3-SWCNTs without acid treatment were prepared using the sol–gel method via a spin coating technique annealed at 450 °C. The optical, morphology and electrical properties of the photoanodes were characterized by means of UV–Vis analysis, atomic force microscopy and field-emission scanning electron microscopy, and J–V curve measurements, respectively. The optical band gap obtained through UV–Vis analysis showed that the acid treatment process modified the band gap of the photoanode, which enhances the V oc of the DSSCs. In addition, In2O3-SWCNTs with acid treatment possess a porous structure that improves the power conversion efficiency (PCE) of the DSSCs. In addition, the diameter of acid-treated SWCNTs was reduced compared to pristine SWCNTs. In2O3-SWCNTs with acid treatment exhibited the highest PCE of 1.40% with J sc of 7.6 mA cm−2, V oc of 0.51 V, and fill factor of 0.36. The increment in V oc is due to the higher band gap obtained through the UV–Vis absorption spectrum. Moreover, In2O3-SWCNTs with acid treatment has a higher electron lifetime with a higher effective diffusion coefficient that slows down the recombination rate and speeds up the electron transport process. (paper)

  9. Solid-phase synthesis of amidine-substituted phenylbenzimidazoles and incorporation of this DNA binding and recognition motif into amino acid and peptide conjugates.

    Garner, Matthew L; Georgiadis, Taxiarchis M; Li, Jessica Bo; Wang, Tianxiu; Long, Eric C

    2014-05-01

    Amidine-substituted phenylbenzimidazoles are well-established DNA-binding structural motifs that have contributed to the development of diverse classes of DNA-targeted agents; this ring system not only assists in increasing the overall DNA affinity of an agent, but can also influence its site selectivity. Seeking a means to conveniently exploit these attributes, a protocol for the on-resin synthesis of amino acid- and peptide-phenylbenzimidazole-amidine conjugates was developed to facilitate installation of phenylbenzimidazole-amidines into peptide chains during the course of standard solid-phase syntheses. Building from a resin-bound amino acid or peptide on Rink amide resin, 4-formyl benzoic acid was coupled to the resin-bound free amine followed by introduction of 3,4-diamino-N'-hydroxybenzimidamide (in the presence of 1,4-benzoquinone) to construct the benzimidazole heterocycle. Finally, the resin-bound N'-hydroxybenzimidamide functionality was reduced to an amidine via 1 M SnCl2·2H2O in DMF prior to resin cleavage to release final product. This procedure permits the straightforward synthesis of amino acids or peptides that are N-terminally capped by a phenylbenzimidazole-amidine ring system. Employing this protocol, a series of amino acid-phenylbenzimidazole-amidine (Xaa-R) conjugates was synthesized as well as dipeptide conjugates of the general form Xaa-Gly-R (where R is the phenylbenzimidazole-amidine and Xaa is any amino acid). PMID:24562478

  10. Polyamine flux analysis by determination of heavy isotope incorporation from 13C, 15N-enriched amino acids into polyamines by LC-MS/MS.

    Cerrada-Gimenez, Marc; Häkkinen, Merja R; Vepsäläinen, Jouko; Auriola, Seppo; Alhonen, Leena; Keinänen, Tuomo A

    2012-02-01

    The study of polyamine flux, i.e. the circulating flow of polyamines through the interconnected biosynthetic and catabolic pathways, is of considerable interest because of the established links between the polyamine metabolism and many diseases, such as cancer and diabetes. To study polyamine flux in detail, a novel method based on following the label incorporation from the (13)C, (15)N-labeled polyamine precursors, arginine, methionine and ornithine, into polyamines by LC-MS/MS was implemented. This methodology was tested on three distinct cell lines with different spermidine/spermine-N (1)-acetyltransferase (SSAT) expression levels, i.e. non-transgenic, transgenic and knockout. These trials allowed the identification of the critical conditions for the successful polyamine flux measurement, such as the functional time frame of label incorporation, until plateau phase with the selected precursor is reached. The novel LC-MS/MS-based method for polyamine flux overcame the limitations of previous existing methodologies, with baseline separation of the different polyamine species and the exact quantification of the incorporated label. Moreover, the obtained results clearly show that the increased SSAT expression is associated with accelerated polyamine flux. PMID:21818565

  11. In vitro model of infected stratum corneum for the efficacy evaluation of poloxamer 407-based formulations of ciclopirox olamine against Trichophyton rubrum as well as differential scanning calorimetry and stability studies.

    Täuber, Anja; Müller-Goymann, Christel C

    2015-10-15

    Superficial fungal skin infections are a common disease and concern 20-25% of the world's population with the dermatophyte Trichophyton rubrum being the main trigger. Due to autoinoculation, fungal skin infections of the feet (tinea pedis) often occur simultaneously with fungal nail infections (onychomycosis). Therefore, the overall objective was the development and characterisation of poloxamer 407-based formulations with the antimycotic active ingredient ciclopirox olamine (CPX) for simultaneous antifungal therapy. The formulations consisted of poloxamer 407, water, isopropyl alcohol, propylene glycol and medium chain triglycerides in given ratios. The in vitro antifungal efficacy against T. rubrum was tested in a novel in vitro model of infected stratum corneum in comparison to a marketed semi-solid formulation containing 1% (w/w) ciclopirox olamine and a marketed nail lacquer containing 8% ciclopirox. Several liquid poloxamer 407-based formulations with only 1% CPX completely inhibited fungal growth after 6 days of incubation, whereas the marketed semi-solid formulation did not inhibit fungal growth. Differential scanning calorimetry studies revealing the interaction between the formulations and the SC showed that increasing isopropyl alcohol/propylene glycol concentrations as well as increasing CPX concentrations caused increasing endothermic transition shifts. Moreover, stability studies at 30 °C exhibited only a slight decrease of the CPX amount after 12 months of storage. Each formulation contained >90% of the initial CPX concentration after termination of the stability studies. PMID:26276254

  12. Efeito da gordura vegetal parcialmente hidrogenada sobre a incorporação de ácidos graxos trans em tecidos de ratos Effect of the partially hydrogenated vegetable fat on the incorporation of trans fatty acids in rat tissues

    Céphora Maria Sabarense

    2003-12-01

    Full Text Available A composição lipídica da dieta pode influenciar o perfil de ácidos graxos dos tecidos. O objetivo do presente estudo foi avaliar a incorporação de ácidos graxos trans no fígado e coração de ratos. Dois grupos com doze ratos Wistar recém-desmamados foram alimentados com duas dietas diferentes por oito semanas. Uma das dietas (experimental foi rica em isômeros trans (33,0% da fração lipídica e apresentou quantidades mínimas de ácidos linoléico e alfa-linolênico (8,0% e 0,7%, respectivamente, da fração lipídica da dieta, enquanto a outra (controle foi nutricionalmente adequada. O perfil de ácidos graxos das dietas e dos tecidos foi avaliado por cromatografia gasosa. Houve incorporação de 14,0% dos ácidos graxos trans no fígado e 8,6% no coração dos animais. Não foi observado efeito inibitório dos ácidos graxos trans no fígado sobre a formação dos ácidos araquidônico e docosahexaenóico. No entanto, no coração houve uma diminuição significante da concentração do ácido docosahexaenóico, provavelmente como reflexo da deficiência de ácido a-linolênico e da incorporação dos trans.The lipid composition of the diet can influence the profile of fatty acids the tissues. The objective of this study was to evaluate the incorporation of trans fatty acids in the liver and in the heart of rats. Two groups with twelve weanling Wistar rats each were fed two different diets for eight weeks. One of the diets (experimental was rich in trans fatty acids (33.0% of total lipids and presented minimal amounts of linoleic and alpha-linolenic acids (8.0% and 0.7%, respectively, of total lipids, while the other (control was nutritionally adequate. The profile of fatty acids of the diets and tissues was evaluated by gas chromatography. There was an incorporation of 14.0% of the trans fatty acids in the liver and 8.6% in the heart. There was no inhibitory effect of the trans fatty acids on the formation of arachidonic and

  13. Synthesis of a Hoechst 32258 analogue amino acid building block for direct incorporation of a fluorescent, high-affinity DNA binding motif into peptides

    Behrens, C; Harrit, N; Nielsen, P E

    2001-01-01

    The synthesis of a new versatile "Hoechst 33258-like" Boc-protected amino acid building block for peptide synthesis is described. It is demonstrated that this new ligand is an effective mimic of Hoechst 33258 in terms of DNA affinity and sequence specificity. Furthermore, this minor groove binder...

  14. Synthesis, structure elucidation, biological screening, molecular modeling and DNA binding of some Cu(II) chelates incorporating imines derived from amino acids

    Abdel-Rahman, Laila H.; Abu-Dief, Ahmed M.; Ismael, Mohammed; Mohamed, Mounir A. A.; Hashem, Nahla Ali

    2016-01-01

    Three tridentate Schiff bases amino acids were prepared by direct condensation of 3-methoxysalicylaldehyde (MS) or 4-diethylaminosalicylaldehyde (DS) with α-amino acid ligands [L-phenylalanine (P), L-histidine (H) and DL-tryptophan (T)]. The prepared Schiff bases amino acids were investigated by melting points, elemental analysis, 1HNMR and 13CNMR, IR, UV-Vis spectra, conductivity and magnetic measurements analyses. Subsequently, copper was introduced and Cu(II) complexes formed. These complexes were analyzed by thermal and elemental analyses and further investigated by FT-IR and UV/Vis spectroscopies. The experimental results indicating that all Cu(II) complexes contain hydrated water molecules (except DSPCu complex) and don't contain coordinated water molecules. The kinetic and thermal parameters were extracted from the thermal data using Coast and Redfern method. The molar conductance values of the Schiff base amino acid ligands and their Cu(II) complexes were relatively low, showing that these compounds have non-electrolytic nature. Magnetic susceptibility measurements showed the diamagnetic nature of the Schiff base amino acid ligands and paramagnetic nature of their complexes. Additionally, a spectrophotometric method was determined to extract their stability constants. It was found that the complexes possess 1:2 (M:L) stoichiometry. The results suggested that 3-methoxysalicylaldehyde and 4-diethylaminosalicylaldehyde amino acid Schiff bases behave as monobasic tridentate ONO ligands and coordinate Cu(II) ions in octahedral geometry according to the general formula [Cu(HL)2]·nH2O. To further understanding the structural and electronic properties of these complexes, Density Functional Theory (DFT) calculations were employed and provided a satisfactory description. The optimized structures of MST Schiff base ligand and its complex were calculated using DFT. The antimicrobial activity of the Schiff base ligands and their complexes were screened against some

  15. Carboxylic acid derivatives via catalytic carboxylation of unsaturated hydrocarbons: whether the nature of a reductant may determine the mechanism of CO2 incorporation?

    Kirillov, E.; Carpentier, J.-F.; Bunel, E

    2015-01-01

    International audience Application of CO2 as a renewable feedstock and C1 building block for prodn. of commodity and fine chems. is a highly challenging but obvious industry-relevant task. Of particular interest is the catalytic coupling of CO2 with inexpensive unsatd. hydrocarbons (olefins, dienes, styrenes, alkynes), providing direct access to carboxylic acids and their derivs. Although not brand new for the scientific community, it is still a complete challenge, as no truly effective ca...

  16. Incorporation of EPA and DHA into plasma phospholipids in response to different omega-3 fatty acid formulations - a comparative bioavailability study of fish oil vs. krill oil

    Schuchardt Jan; Schneider Inga; Meyer Henrike; Neubronner Juliane; von Schacky Clemens; Hahn Andreas

    2011-01-01

    Abstract Background Bioavailability of omega-3 fatty acids (FA) depends on their chemical form. Superior bioavailability has been suggested for phospholipid (PL) bound omega-3 FA in krill oil, but identical doses of different chemical forms have not been compared. Methods In a double-blinded crossover trial, we compared the uptake of three EPA+DHA formulations derived from fish oil (re-esterified triacylglycerides [rTAG], ethyl-esters [EE]) and krill oil (mainly PL). Changes of the FA composi...

  17. Sense codon emancipation for proteome-wide incorporation of noncanonical amino acids: rare isoleucine codon AUA as a target for genetic code expansion

    Bohlke, Nina; Budisa, Nediljko

    2014-01-01

    One of the major challenges in contemporary synthetic biology is to find a route to engineer synthetic organisms with altered chemical constitution. In terms of core reaction types, nature uses an astonishingly limited repertoire of chemistries when compared with the exceptionally rich and diverse methods of organic chemistry. In this context, the most promising route to change and expand the fundamental chemistry of life is the inclusion of amino acid building blocks beyond the canonical 20 ...

  18. 2-ketogluconic acid secretion by incorporation of Pseudomonas putida KT 2440 gluconate dehydrogenase (gad) operon in Enterobacter asburiae PSI3 improves mineral phosphate solubilization.

    Kumar, Chanchal; Yadav, Kavita; Archana, G; Naresh Kumar, G

    2013-09-01

    Enterobacter asburiae PSI3 is known to efficiently solubilize rock phosphate by secretion of approximately 50 mM gluconic acid in Tris-buffered medium in the presence of 75 mM glucose and in a mixture of seven aldosugars each at 15 mM concentration, mimicking alkaline vertisol soils. Efficacy of this bacterium in the rhizosphere requires P release in the presence of low amount of sugars. To achieve this, E. asburiae PSI3 has been manipulated to express gluconate dehydrogenase (gad) operon of Pseudomonas putida KT 2440 to produce 2-ketogluconic acid. E. asburiae PSI3 harboring gad operon had 438 U of GAD activity, secreted 11.63 mM 2-ketogluconic and 21.65 mM gluconic acids in Tris-rock phosphate-buffered medium containing 45 mM glucose. E. asburiae PSI3 gad transformant solubilized 0.84 mM P from rock phosphate in TRP-buffered liquid medium. In the presence of a mixture of seven sugars each at 12 mM, the transformant brought about a drop in pH to 4.1 and released 0.53 mM P. PMID:23666029

  19. Influence of hydroxyurea on nucleic acids content and 3H-uridine incorporation in callus and tumorous tobacco tissues cultured in vitro

    A. Bielecka

    2015-05-01

    Full Text Available In callus and tumor tissues of Nicotiana tabacum cultured for 39 days in media supplemented with various concentrations of hydroxyurea (1.3 x 10-4 M - 1.3 x 10-3 M a decrease of DNA content (ca. 24 per cent in callus tissue and ca. 23 per cent in tumour tissue and a decrease of RNA content (over 10 per cent and ca. 9 per cent in callus and tumour tissue, respectively was observed. The autoradiographic method showed that a long-lasting action of this com-pound inhibits RNA synthesis. A stronger inhibitory influence of hydroxyurea upon incorporation of 3H-uridine from the incubation medium was revealed.

  20. Incorporation and profile of fatty acids in tilapia fillets (Oreochromis niloticus) fed with tung oil Incorporação e perfil de ácidos graxos em filés de tilápia (Oreochromis niloticus) alimentada com óleo de tungue

    Elton Guntendorfer Bonafé; Damila Rodrigues de Morais; Luana Caroline de Figueiredo; Nilson Evelázio de Souza; Oscar Oliveira Santos; Thiago Claus; Jesuí Vergílio Visentainer

    2013-01-01

    The acceptance of tung oil enriched diet and the incorporation of conjugated linolenic acid - CLnA into fillets of Genetically Improved Farmed Tilapia (GIFT) were investigated. The diet was well accepted, and after 10 days CLnA was incorporated into the fillets with a 1.02% content of total fatty acids (FA). In addition, biosynthesis of the conjugated linoleic acid isomers - CLA (0.31% of fillet total FA content) from CLnA, and the presence of alpha-linolenic acid - LNA (1.08% of fillet total...

  1. Feeding laying hens seal blubber oil: effects on egg yolk incorporation, stereospecific distribution of omega-3 fatty acids, and sensory aspects.

    Schreiner, M; Hulan, H W; Razzazi-Fazeli, E; Böhm, J; Iben, C

    2004-03-01

    Seventy-two 26-wk-old Single Comb White Leghorn laying hens were randomly assigned to 36 cages (2 per cage) in a 3-orthogonal 4 x 4 latin square, with the fourth row suppressed, to assess the effect of feeding refined seal blubber oil (SBO, containing 22.2% omega-3 fatty acids) on the fatty acid composition and position in the egg yolk lipids. The experiment was conducted over a period of 9 wk. Eggs were collected and numbered, and the weights were recorded for each week and cage. Eggs collected at wk 5 and 9 were used for total lipid, lipid class, fatty acid, and positional analyses. Sensory evaluation was carried out on eggs collected at wk 6 and 7. Feeding SBO at 1.25% led to an increase (P 0.1) on the sensory attributes of the egg and on production parameters such as egg weight, number of eggs laid, and feed intake (P > 0.05). When feeding SBO in amounts higher than 1.25% proportionately, a plateau effect of the LCn3PUFA content of the eggs was observed. This appears to be because the PUFA content in the sn-2 position of the phospholipids cannot exceed a certain amount. When this amount is reached, the LCn3PUFA will be increasingly stored in triglycerides. The results presented here clearly indicate how eggs can be produced with optimized composition of LCn3PUFA without affecting (P > 0.1) the sensory properties of the eggs. The procedures elaborated herein provide directly applicable consequences for the food industry. PMID:15049501

  2. Identification of protein-protein binding sites by incorporating the physicochemical properties and stationary wavelet transforms into pseudo amino acid composition.

    Jia, Jianhua; Liu, Zi; Xiao, Xuan; Liu, Bingxiang; Chou, Kuo-Chen

    2016-09-01

    With the explosive growth of protein sequences entering into protein data banks in the post-genomic era, it is highly demanded to develop automated methods for rapidly and effectively identifying the protein-protein binding sites (PPBSs) based on the sequence information alone. To address this problem, we proposed a predictor called iPPBS-PseAAC, in which each amino acid residue site of the proteins concerned was treated as a 15-tuple peptide segment generated by sliding a window along the protein chains with its center aligned with the target residue. The working peptide segment is further formulated by a general form of pseudo amino acid composition via the following procedures: (1) it is converted into a numerical series via the physicochemical properties of amino acids; (2) the numerical series is subsequently converted into a 20-D feature vector by means of the stationary wavelet transform technique. Formed by many individual "Random Forest" classifiers, the operation engine to run prediction is a two-layer ensemble classifier, with the 1st-layer voting out the best training data-set from many bootstrap systems and the 2nd-layer voting out the most relevant one from seven physicochemical properties. Cross-validation tests indicate that the new predictor is very promising, meaning that many important key features, which are deeply hidden in complicated protein sequences, can be extracted via the wavelets transform approach, quite consistent with the facts that many important biological functions of proteins can be elucidated with their low-frequency internal motions. The web server of iPPBS-PseAAC is accessible at http://www.jci-bioinfo.cn/iPPBS-PseAAC , by which users can easily acquire their desired results without the need to follow the complicated mathematical equations involved. PMID:26375780

  3. Attempting to monitor the incorporation of deuterium into indole-3-acetic acid and tryptophan in Zea mays grown on deuterium oxide labeled water

    We are attempting to determine when seedlings of Zea mays sweet corn, var. Silver Queen begin de novo biosynthesis of tryptophan and indole-3-acetic acid (IAA). We wish to use the general precursor, deuterium labeled water, to minimize assumptions as to the biosynthetic route. Protium in positions 2, 4, 5, 6 ampersand 7 of the indole ring are non-exchangeable. IAA and tryptophan synthesized via the shikimic acid pathway would contain deuterium in one or more of these positions . The protium on the indene nitrogen, the carboxyl, the amino group, or the protium alpha to the carboxyl exchange readily and so are removed prior to analysis by base catalyzed exchange. The IAA, or trypotophan, is then purified by DEAE, Dowex 50, and two HPLC steps. IAA is methylated with diazomethane and analyzed by GC/MS. Trytophan is acetylated with triethylamine-acetic anhydride and then methylated with diazomethane and analyzed by GC/MS. Results of these studies on plants grown for varying lengths of time and under various light and nutrient conditions will be reported

  4. Probing the Active Site of MIO-dependent Aminomutases, Key Catalysts in the Biosynthesis of amino Acids Incorporated in Secondary Metabolites

    Cooke, H.; Bruner, S

    2010-01-01

    The tyrosine aminomutase SgTAM produces (S)-{beta}-tyrosine from L-tyrosine in the biosynthesis of the enediyne antitumor antibiotic C-1027. This conversion is promoted by the methylideneimidazole-5-one (MIO) prosthetic group. MIO was first identified in the homologous family of ammonia lyases, which deaminate aromatic amino acids to form {alpha},{beta}-unsaturated carboxylates. Studies of substrate specificity have been described for lyases but there have been limited reports in altering the substrate specificity of aminomutases. Furthermore, it remains unclear as to what structural properties are responsible for catalyzing the presumed readdition of the amino group into the {alpha},{beta}-unsaturated intermediates to form {beta}-amino acids. Attempts to elucidate specificity and mechanistic determinants of SgTAM have also proved to be difficult as it is recalcitrant to perturbations to the active site via mutagenesis. An X-ray cocrystal structure of the SgTAM mutant of the catalytic base with L-tyrosine verified important substrate binding residues as well as the enzymatic base. Further mutagenesis revealed that removal of these crucial interactions renders the enzyme inactive. Proposed structural determinants for mutase activity probed via mutagenesis, time-point assays and X-ray crystallography revealed a complicated role for these residues in maintaining key quaternary structure properties that aid in catalysis.

  5. Probing the active site of MIO-dependent aminomutases, key catalysts in the biosynthesis of beta-amino acids incorporated in secondary metabolites.

    Cooke, Heather A; Bruner, Steven D

    2010-09-01

    The tyrosine aminomutase SgTAM produces (S)-ss-tyrosine from L-tyrosine in the biosynthesis of the enediyne antitumor antibiotic C-1027. This conversion is promoted by the methylideneimidazole-5-one (MIO) prosthetic group. MIO was first identified in the homologous family of ammonia lyases, which deaminate aromatic amino acids to form alpha,ss-unsaturated carboxylates. Studies of substrate specificity have been described for lyases but there have been limited reports in altering the substrate specificity of aminomutases. Furthermore, it remains unclear as to what structural properties are responsible for catalyzing the presumed readdition of the amino group into the alpha,ss-unsaturated intermediates to form ss-amino acids. Attempts to elucidate specificity and mechanistic determinants of SgTAM have also proved to be difficult as it is recalcitrant to perturbations to the active site via mutagenesis. An X-ray cocrystal structure of the SgTAM mutant of the catalytic base with L-tyrosine verified important substrate binding residues as well as the enzymatic base. Further mutagenesis revealed that removal of these crucial interactions renders the enzyme inactive. Proposed structural determinants for mutase activity probed via mutagenesis, time-point assays and X-ray crystallography revealed a complicated role for these residues in maintaining key quaternary structure properties that aid in catalysis. PMID:20577998

  6. Predicting lysine phosphoglycerylation with fuzzy SVM by incorporating k-spaced amino acid pairs into Chou׳s general PseAAC.

    Ju, Zhe; Cao, Jun-Zhe; Gu, Hong

    2016-05-21

    As a new type of post-translational modification, lysine phosphoglycerylation plays a key role in regulating glycolytic process and metabolism in cells. Due to the traditional experimental methods are time-consuming and labor-intensive, it is important to develop computational methods to identify the potential phosphoglycerylation sites. However, the prediction performance of the existing phosphoglycerylation site predictor is not satisfactory. In this study, a novel predictor named CKSAAP_PhoglySite is developed to predict phosphoglycerylation sites by using composition of k-spaced amino acid pairs and fuzzy support vector machine. On the one hand, after many aspects of assessments, we find the composition of k-spaced amino acid pairs is more suitable for representing the protein sequence around the phosphoglycerylation sites than other encoding schemes. On the other hand, the proposed fuzzy support vector machine algorithm can effectively handle the imbalanced and noisy problem in phosphoglycerylation sites training dataset. Experimental results indicate that CKSAAP_PhoglySite outperforms the existing phosphoglycerylation site predictor Phogly-PseAAC significantly. A matlab software package for CKSAAP_PhoglySite can be freely downloaded from https://github.com/juzhe1120/Matlab_Software/blob/master/CKSAAP_PhoglySite_Matlab_Software.zip. PMID:26908349

  7. Two-photon absorption of fluorescent protein chromophores incorporating non-canonical amino acids: TD-DFT screening and classical dynamics.

    Alaraby Salem, M; Brown, Alex

    2015-10-14

    Two-photon spectroscopy of fluorescent proteins is a powerful bio-imaging tool characterized by deep tissue penetration and little damage. However, two-photon spectroscopy has lower sensitivity than one-photon microscopy alternatives and hence a protein with a large two-photon absorption cross-section is needed. We use time-dependent density functional theory (TD-DFT) at the B3LYP/6-31+G(d,p) level of theory to screen twenty-two possible chromophores that can be formed upon replacing the amino-acid Tyr66 that forms the green fluorescent protein (GFP) chromophore with a non-canonical amino acid. A proposed chromophore with a nitro substituent was found to have a large two-photon absorption cross-section (29 GM) compared to other fluorescent protein chromophores as determined at the same level of theory. Classical molecular dynamics are then performed on a nitro-modified fluorescent protein to test its stability and study the effect of the conformational flexibility of the chromophore on its two-photon absorption cross-section. The theoretical results show that the large cross-section is primarily due to the difference between the permanent dipole moments of the excited and ground states of the nitro-modified chromophore. This large difference is maintained through the various conformations assumed by the chromophore in the protein cavity. The nitro-derived protein appears to be very promising as a two-photon absorption probe. PMID:26370051

  8. Techno-economic analysis for incorporating a liquid-liquid extraction system to remove acetic acid into a proposed commercial scale biorefinery.

    Aghazadeh, Mahdieh; Engelberth, Abigail S

    2016-07-01

    Mitigating the effect of fermentation inhibitors in bioethanol plants can have a great positive impact on the economy of this industry. Liquid-liquid extraction (LLE) using ethyl acetate is able to remove fermentation inhibitors-chiefly, acetic acid-from an aqueous solution used to produce bioethanol. The fermentation broth resulting from LLE has higher performance for ethanol yield and its production rate. Previous techno-economic analyses focused on second-generation biofuel production did not address the impact of removing the fermentation inhibitors on the economic performance of the biorefinery. A comprehensive analysis of applying a separation system to mitigate the fermentation inhibition effect and to provide an analysis on the economic impact of removal of acetic acid from corn stover hydrolysate on the overall revenue of the biorefinery is necessary. This study examines the pros and cons associated with implementing LLE column along with the solvent recovery system into a commercial scale bioethanol plant. Using details from the NREL-developed model of corn stover biorefinery, the capital costs associated with the equipment and the operating cost for the use of solvent were estimated and the results were compared with the profit gain due to higher ethanol production. Results indicate that the additional capital will add 1% to the total capital and manufacturing cost will increase by 5.9%. The benefit arises from the higher ethanol production rate and yield as a consequence of inhibitor extraction and results in a $0.35 per gallon reduction in the minimum ethanol selling price (MESP). © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:971-977, 2016. PMID:27390294

  9. Enhanced adsorptive and photocatalytic achievements in removal of methylene blue by incorporating tungstophosphoric acid-TiO2 into MCM-41

    The use of titania-dispersed materials in photocatalytic processes has been proposed as an alternative to the conventional bare TiO2, in order to modify the surface area and activity of the catalyst. A homogeneously dispersed Keggin unit into TiO2 was synthesized using tungstophosphoric acid (TPA) and titanium tetraisopropoxide. This compound was then loaded into MCM-41 by dispersing it in a suspension containing the mesoporous phase. Two other titanium-containing MCM-41 catalysts, Ti-MCM-41 and TiO2/MCM-41 were also prepared using isomorphous substitution synthesis method and impregnation method, respectively, for the sake of comparison. The prepared photocatalysts were characterized by X-ray diffraction (XRD), nitrogen physisorption (BET) and chemical analysis. The catalysts were used to study degradation of methylene blue (MB) in aqueous solution. XRD result shows a pure anatase crystalline phase for TPA-containing TiO2 indicating that there is good molecular distribution of tungstophosphoric acid into TiO2 structure. Supported TPA-TiO2 into MCM-41 shows both TPA-TiO2 and MCM-41 characteristic X-ray reflections in the high-angle and low-angle parts of the XRD patterns, respectively. The experimental results show that adsorption is a major constituent in the elimination of MB from the dye solutions by the TPA-containing materials. Exploitation of both adsorption and photocatalytic processes speeds up the removal of the dye using the TPA-TiO2-loaded MCM-41 photocatalyst. The elimination of MB is completed within 15 min for a 30 mg l-1 MB solution containing a catalyst dose of 100 mg/100 ml. The efficiencies of the other photocatalysts such as commercial TiO2, Ti-MCM-41, TiO2/MCM-41 and TPA-TiO2 for adsorption and degradation of MB were also studied and compared with that of the prepared catalyst.

  10. Synthesis, structure and thermochemical study of a cobalt energetic coordination compound incorporating 3,5-diamino-1,2,4-triazole and pyridine-2,6-dicarboxylic acid

    Highlights: • An energetic coordination compound with dinuclear cobalt unit has been synthesized and structurally characterized. • The standard molar enthalpy of formation of the compound was determined by a designed thermochemical cycle. • The specific heat capacity of the compound at T = 298.15 K was determined to be 1.13 ± 0.02 J · mol−1 · K−1. - Abstract: An energetic coordination compound [Co2(C2H5N5)2(C7H3NO4)2(H2O)2]·2H2O (Hdatrz(C2H5N5) = 3,5-diamino-1,2,4-triazole, H2pda(C7H5NO4) = pyridine-2,6-dicarboxylic acid) has been synthesized and characterized by elemental analysis, chemical analysis, IR spectroscopy, single-crystal X-ray diffraction and thermal analysis. X-ray diffraction analysis confirmed that the compound possessed a di-nuclear unit and featured a 3D super-molecular structure. Furthermore, a reasonable thermochemical cycle was designed based on the preparation reaction of the compound and the standard molar enthalpy of dissolution of reactants and products was measured by the RD496-2000 calorimeter. Finally, the standard molar enthalpy of formation of the compound was determined to be −(2475.0 ± 3.1) kJ · mol−1 in accordance with Hess’s law. In addition, the specific heat capacity of the compound at T = 298.15 K was determined to be (1.13 ± 0.02) J · K−1 · g−1 by RD496-2000 calorimeter

  11. Effects of butter high in ruminant trans and monounsaturated fatty acids on lipoproteins, incorporation of fatty acids into lipid classes, plasma C-reactive protein, oxidative stress, hemostatic variables, and insulin in healthy young men

    Tholstrup, T.; Raff, M.; Basu, S.; Nonboe, P.; Sejrsen, K.; Straarup, Ellen Marie

    2006-01-01

    Background: Evidence suggests that ruminant trans fatty acids (FAs), such as vaccenic acid, do not increase the risk of ischemic heart disease (IHD). However, the effects of ruminant trans FAs on risk markers of IHD have been poorly investigated. Objective: The objective was to investigate the...... effect of butter with a naturally high content of vaccenic acid and a concomitantly higher content of monounsaturated FAs on classic and novel risk markers of IHD. Design: In a double-blind, randomized, 5-wk, parallel intervention study, 42 healthy young men were given 115 g fat/d from test butter that...... observed. Conclusions: Butter high in ruminant trans and monounsaturated FAs resulted in significantly lower total and HDL cholesterol than did the control butter with higher amounts of saturated FAs. It may be that the differences were due to the greater content of monounsaturated FAs and the lesser...

  12. Effect of Temperature-Sensitive Poloxamer Solution/Gel Material on Pericardial Adhesion Prevention: Supine Rabbit Model Study Mimicking Cardiac Surgery.

    Hyun Kang

    Full Text Available We investigated the mobility of a temperature-sensitive poloxamer/Alginate/CaCl2 mixture (PACM in relation to gravity and cardiac motion and the efficacy of PACM on the prevention of pericardial adhesion in a supine rabbit model.A total of 50 rabbits were randomly divided into two groups according to materials applied after epicardial abrasion: PACM and dye mixture (group PD; n = 25 and saline as the control group (group CO; n = 25. In group PD, rabbits were maintained in a supine position with appropriate sedation, and location of mixture of PACM and dye was assessed by CT scan at the immediate postoperative period and 12 hours after surgery. The grade of adhesions was evaluated macroscopically and microscopically two weeks after surgery.In group PD, enhancement was localized in the anterior pericardial space, where PACM and dye mixture was applied, on immediate post-surgical CT scans. However, the volume of the enhancement was significantly decreased at the anterior pericardial space 12 hours later (P < .001. Two weeks after surgery, group PD had significantly lower macroscopic adhesion score (P = .002 and fibrosis score (P = .018 than did group CO. Inflammation score and expression of anti-macrophage antibody in group PD were lower than those in group CO, although the differences were not significant.In a supine rabbit model study, the anti-adhesion effect was maintained at the area of PACM application, although PACM shifted with gravity and heart motion. For more potent pericardial adhesion prevention, further research and development on the maintenance of anti-adhesion material position are required.

  13. Poloxamer 407-induced atherosclerosis in mice appears to be due to lipid derangements and not due to its direct effects on endothelial cells and macrophages

    Thomas P. Johnston

    2003-01-01

    Full Text Available Coronary heart disease secondary to atherosclerosis is still the leading cause of death in the US. Animal models used for elucidating the pathogenesis of this disease primarily involve rabbits and pigs. Previous studies from this laboratory have demonstrated intraperitoneal injections of poloxamer 407 (P-407 in both male and female mice will lead to hyperlipidemia and atherosclerosis, suggesting the use of this polymer to develop a mouse model of atherosclerosis. In order to understand the mechanism of P-407-induced hyperlipidemia and vascular lesion formation, we evaluated the direct effects of P-407 on endothelial cell and macrophage functions in vitro, and its in vivo effects on the oxidation of circulating lipids following long-term (4 month administration. Our results demonstrated that incubation of P-407 with human umbilical vein endothelial cells in culture did not influence either cell proliferation or interleukin-6 and interleukin-8 production over a concentration range of 0-40 μM. In addition, nitric oxide production by macrophages was not affected by P-407 over a concentration range of 0-20 μM. Finally, we demonstrated that while P-407 could not induce the oxidation of LDL-C in vitro, long-term (4 month administration of P-407 in mice resulted in elevated levels of oxidized lipids in the plasma. Thus, it is suggested that the formation of atherosclerotic lesions in this mouse model of atherosclerosis does not result from either direct stimulation of endothelial cells or macrophage activation by P-407. Instead, these data would support the premise that oxidation of lipids (perhaps low-density lipoprotein cholesterol by an indirect mechanism following injection of P-407 may represent one of the mechanisms responsible for atheroma formation.

  14. Correção da acidez do solo em função de modos de incorporação de calcário Correction of soil acidity in function of lime incorporation manners

    Pedro Henrique Weirich Neto

    2000-04-01

    Full Text Available A solubilização do calcário no solo, sem o que não ocorre neutralização da acidez, demanda tempo e exige incorporação para aumentar o contato entre o corretivo e os colóides do solo. Com o objetivo de avaliar a correção da acidez do solo em função de diferentes modos de incorporação de calcário, foi conduzido um experimento, no ano agrícola 1993/94, em um Latossolo vermelho-escuro, distrófico, de campo nativo, do município de Ponta Grossa (PR. O delineamento experimental empregado foi o de blocos ao acaso, em parcelas subdivididas, com três repetições. Os tratamentos foram constituídos por cinco modos de incorporação: arado de discos mais duas gradagens, grade aradora mais duas gradagens, enxada rotativa, arado escarificador mais duas gradagens e distribuição na superfície sem incorporação, e por quatro doses de calcário dolomítico: 0, 2,8, 6,6 e 10,3 t ha-1, visando a elevar a saturação por bases do solo a 30, 60 e 90%. A planta teste utilizada foi o milho. Verificou-se que o modo de incorporação do corretivo no solo afeta a sua eficiência em relação à profundidade no perfil, na qual há neutralização da acidez. Houve elevação da saturação por bases do solo até a camada de 20cm, 15cm e 10cm, respectivamente, para os seguintes modos de incorporação: enxada rotativa, arado de discos ou grade aradora e arado escarificador ou calcário distribuído na superfície sem incorporação. Os valores de saturação por bases desejados não foram alcançados, três meses após a calagem, independente do modo de incorporação de calcário.The solubilization of lime in soil, without which the acidity neutralization does not occur, demands time and requires incorporation for increasing the contact between lime and soil coloides. In order to study the correction of soil acidity in function of lime incorporation manners, a field experiment was carried out, in Ponta Grossa, State of Paraná, Brazil, in a

  15. Tuning the structures of three coordination polymers incorporating ZnII and 2,2‧-dichloro-4,4‧-azodibenzoic acid via selective auxiliary ligands

    Zeng, Xiao-Ping; Ming, Mei

    2015-11-01

    By tuning the auxiliary ligands in the assembling reaction, three ZnII coordination polymers of [Zn(Cl-adc) (phen) (H2O)](DMF) (1), [Zn(Cl-adc) (DMA)](DMA) (2), and [Zn(Cl-adc) (dip)](DMF)0.5 (3) (Cl-H2adc = 2,2‧-dichloro-4,4‧-azodibenzoic acid, phen = 1,10-phenanthroline, dip = 1,3-di(imidazole)propane) have been successfully synthesized and characterized by single-crystal X-ray diffraction study, elemental analysis, IR spectra, TGA analyses, solid-state fluorescent property, and powder X-ray diffraction (PXRD). Single crystal X-ray diffraction reveals that 1 and 2 displays a 1D polymeric chain and 2D sql layered net with the presence of chelated phen and terminal DMA ligands, respectively. By incorporating dip linker, 3 exhibits a 2D + 2D → 3D entangled network, with each 2D net portraying wavelike sql layered structure. Their structural divergences should be properly attributed to fact that, the structural topologies can be well regulated by using three auxiliary ligands incorparating different coordination function.

  16. Practical preparation procedures for docetaxel-loaded nanoparticles using polylactic acid-co-glycolic acid

    Keum CG

    2011-10-01

    Full Text Available Chang-Gu Keum1*, Young-Wook Noh1*, Jong-Suep Baek1, Ji-Ho Lim1, Chan-Ju Hwang1, Young-Guk Na1, Sang-Chul Shin2, Cheong-Weon Cho11College of Pharmacy and Institute of Drug Research and Development, Chungnam National University, Gungdong, Yuseonggu, Daejeon, South Korea; 2College of Pharmacy, Chonnam National University, Yongbongdong, Buggu, Gwangju, South Korea *These authors contributed equally to this work Background: Nanoparticles fabricated from the biodegradable and biocompatible polymer, polylactic-co-glycolic acid (PLGA, are the most intensively investigated polymers for drug delivery systems. The objective of this study was to explore fully the development of a PLGA nanoparticle drug delivery system for alternative preparation of a commercial formulation. In our nanoparticle fabrication, our purpose was to compare various preparation parameters. Methods: Docetaxel-loaded PLGA nanoparticles were prepared by a single emulsion technique and solvent evaporation. The nanoparticles were characterized by various techniques, including scanning electron microscopy for surface morphology, dynamic light scattering for size and zeta potential, x-ray photoelectron spectroscopy for surface chemistry, and high-performance liquid chromatography for in vitro drug release kinetics. To obtain a smaller particle, 0.2% polyvinyl alcohol, 0.03% D-α-tocopheryl polyethylene glycol 1000 succinate (TPGS, 2% Poloxamer 188, a five-minute sonication time, 130 W sonication power, evaporation with magnetic stirring, and centrifugation at 8000 rpm were selected. To increase encapsulation efficiency in the nanoparticles, certain factors were varied, ie, 2–5 minutes of sonication time, 70–130 W sonication power, and 5–25 mg drug loading. Results: A five-minute sonication time, 130 W sonication power, and a 10 mg drug loading amount were selected. Under these conditions, the nanoparticles reached over 90% encapsulation efficiency. Release kinetics showed that 20

  17. Bilateral lesions of suprachiasmatic nuclei affect circadian rhythms in [3H]-thymidine incorporation into deoxyribonucleic acid in mouse intestinal tract, mitotic index of corneal epithelium, and serum corticosterone

    Investigations into the role of the suprachiasmatic nuclei (SCN) in the coordination of circadian rhythms have presented differing results. Several reports have shown that ablation of the suprachiasmatic nuclei (SCNA) alters the phase and amplitude of rhythms but does not abolish them. The present study investigates the effect of SCNA on the rhythms in cell proliferation in various regions of the intestinal tract as measured by the incorporation of [3H]-thymidine into deoxyribonucleic acid, in the mitotic activity of the corneal epithelium, and in serum corticosterone levels. The study involved mice with verified lesions of the SCN (six to 13 mice per time point) and control groups of both sham-operated and unoperated mice (seven of each per time point). The mice were killed in groups that represented seven time points over a single 24 hr span (3 hr intervals with the 0800 hr sampled both at start and end of the series). The tissues examined were the tongue, esophagus, gastric stomach, and colon for DNA synthesis, the corneal epithelium for mitotic index, and blood serum for corticosterone level. The most consistent result of SCNA was a phase advance in the rhythms in cell proliferation in the tongue, esophagus, gastric stomach, colon, and corneal epithelium. A reduction in rhythm amplitude occurred in the tongue, esophagus, and corneal epithelium; however, there was an amplitude increase for the stomach, colon, and serum corticosterone. The mesor (rhythm-adjusted mean) was increased by SCNA in all tissues except the corneal epithelium. These findings further support the role of the suprachiasmatic nuclear area in the control of rhythms in cell proliferation and corticosterone production, by acting as a ''phase-resetter'' and as a modulator of rhythm amplitude

  18. Incorporação de resíduo do tratamento de drenagem ácida em massa de cerâmica vermelha Incorporation of acid drainage treatment sludge waste into red ceramic products

    S. L. Galatto

    2009-03-01

    Full Text Available Neste trabalho, apresenta-se o estudo experimental em escala laboratorial e industrial da incorporação de resíduo do tratamento de Drenagem Ácida de Mina (DAM na formulação de massa de cerâmica vermelha, com substituição parcial das argilas. Foram realizados algumas análises e ensaios nas amostras dos blocos cerâmicos: análise dilatométrica a verde, absorção de água, resistência à compressão, eflorescência de sais solúveis e emissões atmosféricas (MP e SOx. Os resultados das análises nos corpos de prova ensaiados indicam aumento de resistência a compressão. Os ensaios de eflorescência de sais solúveis indicam maior intensidade de cristalização de sais solúveis na superfície dos corpos de prova e blocos cerâmicos à medida que se aumenta o percentual de resíduo. As análises realizadas nos blocos cerâmicos ensaiados em escala industrial, recomendam uma proporção de até 2,5% de resíduo na massa cerâmica.An experimental laboratory and industry scale study of the incorporation of residue from Acid Drainage Mine (ADM in the ceramics mass, with partial substitution of clays, is presented. Some analyses and assays of the ceramic blocks were been carried out: dilatometry, water absorption, compressive strength, efflorescence and emissions (MP and SOx. The results show an increase of compressive strength. The efflorescence assays indicate greater intensity of crystallization of soluble salts in the surface of the ceramic blocks for increasing residue content. The analyses in the ceramic blocks in industrial scale recommend a ratio of up to 2.5% of waste in bulk ceramics.

  19. Incorporating Feminist Standpoint Theory

    Ahlström, Kristoffer

    2005-01-01

    As has been noted by Alvin Goldman, there are some very interesting similarities between his Veritistic Social Epistemology (VSE) and Sandra Harding’s Feminist Standpoint Theory (FST). In the present paper, it is argued that these similarities are so significant as to motivate an incorporation of...

  20. Staphylococcus aureus Sortase A-Mediated Incorporation of Peptides: Effect of Peptide Modification on Incorporation.

    Silvie Hansenová Maňásková

    Full Text Available The endogenous Staphylococcus aureus sortase A (SrtA transpeptidase covalently anchors cell wall-anchored (CWA proteins equipped with a specific recognition motif (LPXTG into the peptidoglycan layer of the staphylococcal cell wall. Previous in situ experiments have shown that SrtA is also able to incorporate exogenous, fluorescently labelled, synthetic substrates equipped with the LPXTG motif (K(FITCLPETG-amide into the bacterial cell wall, albeit at high concentrations of 500 μM to 1 mM. In the present study, we have evaluated the effect of substrate modification on the incorporation efficiency. This revealed that (i by elongation of LPETG-amide with a sequence of positively charged amino acids, derived from the C-terminal domain of physiological SrtA substrates, the incorporation efficiency was increased by 20-fold at 10 μM, 100 μM and 250 μM; (ii Substituting aspartic acid (E for methionine increased the incorporation of the resulting K(FITCLPMTG-amide approximately three times at all concentrations tested; (iii conjugation of the lipid II binding antibiotic vancomycin to K(FITCLPMTG-amide resulted in the same incorporation levels as K(FITCLPETG-amide, but much more efficient at an impressive 500-fold lower substrate concentration. These newly developed synthetic substrates can potentially find broad applications in for example the in situ imaging of bacteria; the incorporation of antibody recruiting moieties; the targeted delivery and covalent incorporation of antimicrobial compounds into the bacterial cell wall.

  1. Thermosetting gel for the delivery of 5-aminolevulinic acid esters to the cervix.

    Collaud, Sabine; Peng, Qian; Gurny, Robert; Lange, Norbert

    2008-07-01

    5-Aminolevulinic acid (5-ALA)-mediated photodynamic therapy has been proposed as an alternative, cervix-sparing treatment for cervical intraepithelial neoplasia (CIN). In this context, topical application of 5-ALA to the cervix is beneficial due to the small necessary dose and its minimal side effects. Therefore, lipophilic 5-ALA esters, such as hexylaminolevulinate (HAL), have led to improved local bioavailability and therapeutic efficacy. Hydrogels have shown to be more appropriate for the local delivery of these derivatives, but due to the limited long-term stability of such formulations at 25 degrees C, the development of an extemporaneously prepared hydrogel targeting CIN can be advantageous. Therefore, a poloxamer 407 thermosetting gel, which is liquid at room temperature and becomes a semi-solid when in contact with the female genital tract, has been evaluated in vitro and in vivo. Rheological evaluation has shown that a 17.0% poloxamer 407 hydrogel with a sol-gel transition at 24.8 +/- 0.6 degrees C was the best formulation for easy application and optimal residence time. Furthermore, similarly to other hydrogels previously tested, such a formulation shows a more complete HAL release in vitro than conventional cream vehicles, and tends to increase porphyrin accumulation in nude mice skin. Finally, in vitro release profiles were correlated to the in vivo results. PMID:17828758

  2. Selective incorporation of 5-hydroxytryptophan into proteins in mammalian cells

    Zhang, Zhiwen; Alfonta, Lital; Schultz, Peter G

    2014-02-25

    This invention provides methods and compositions for incorporation of an unnatural amino acid into a peptide using an orthogonal aminoacyl tRNA synthetase/tRNA pair. In particular, an orthogonal pair is provided to incorporate 5-hydroxy-L-tryptophan in a position encoded by an opal mutation.

  3. Deuterium incorporation into Escherichia-coli proteins

    Lederer, H.; May, R. P.; Kjems, Jørgen;

    1986-01-01

    -dependent RNA polymerase of Escherichia coli match when RNA polymerase is isolated from cells grown in a medium containing 46% D2O and unlabelled glucose as carbon source. Their contrasts vanish simultaneously in a dialysis buffer containing 65% D2O. An expression was evaluated which allows the calculation of...... the degree of deuteration and match point of any E. coli protein from the D2O content of the growth medium, taking the 2H incorporation into RNA polymerase amino acids to be representative for all amino acids in E. coli proteins. The small-angle scattering results, on which the calculation of the...

  4. Study on Preparation and Dissolution Rate of Icaritin-Poloxamer 188 Solid Dispersion%淫羊藿素-泊洛沙姆188固体分散体的制备及溶出度研究

    吴玲

    2015-01-01

    OBJECTIVE:To prepare icaritin-poloxamer 188 solid dispersions in order to increase the dissolution rate of icaritin. METHODS:With poloxamer 188 as the carrier,melting method was used to prepare solid dispersions. By comparing the in vitro dissolution rates,the effects of the content of poloxamer 188(the ratios of icaritin to poloxamer 188 were 5∶1,3∶1,2∶1,1∶1,1∶3,1∶5,1∶7,1∶9,1∶11,1∶13,1∶15,1∶17,1∶19,1∶27 and 1∶31),melting temperature(60,70 and 80 ℃)and cooling tem-perature(-20,0 and 20 ℃)on the dissolution rate of icaritin in the solid dispersions were investigated,and the in vitro dissolu-tion rates of icaritin in its active pharmaceutical ingredient,physical mixture and solid dispersions were compared to confirm the for-mation of the solid dispersions. RESULTS:The dissolution rate of icaritin in the prepared solid dispersions increased to some extent as the proportion of the carrier increased. When the ratio of icaritin to the carrier was 1∶17-1∶27,the dissolution rate of icaritin at 120 min was above 90%. Where melting temperature and cooling temperature were respectively determined as 60 ℃ and 0 ℃ after comprehensive comparison,the dissolution rate of icaritin in the solid dispersions was 1.5 times as much as that in the physical mix-ture at 30 min. CONCLUSIONS:The prepared solid dispersion has a significantly higher dissolution rate of icaritin.%目的:制备淫羊藿素-泊洛沙姆188固体分散体以提高淫羊藿素的溶出度。方法:以泊洛沙姆188为载体,采用熔融法制备固体分散体。通过体外溶出度比较,考察了载体泊洛沙姆188的用量(淫羊藿素-泊洛沙姆188质量之比为5∶1、3∶1、2∶1、1∶1、1∶3、1∶5、1∶7、1∶9、1∶11、1∶13、1∶15、1∶17、1∶19、1∶27、1∶31)、熔融温度(60、70、80℃)、冷却温度(-20、0、20℃)对固体分散体中淫羊藿溶出度的影响;同时比较了淫羊藿素原料药及其

  5. Swelling and Drug Release Characteristics of Poly (methacrylic acid-co-poloxamer) hydrogels

    XUHui; LINYan-nan; DINGPing-tian; TIANMei-juan; ZHENGJun-min

    2003-01-01

    Poly (methacrylie acid co-poloxamer) hydrogel networks were synthesized by free-radical solution polymerization, and the dynamic swelling and in vitro release properties of model drugs, dextromethorphan hydrobromide (DMP) and vitamin B12 (VB12) were studied. These gels exhibited pH-dependant swelling and sustained drug release properties, and the water uptake rate and drug release rate in neutral or basic media were higher than that in acidic media. The results showed that the water uptake followed non-Fickian or zero order process in neutral or basic media, and the release of model drugs from hydrogels of appropriate composition was of zero order kinetics over a period of several hours.

  6. Aniline incorporated silica nanobubbles

    M J Rosemary; V Suryanarayanan; Ian Maclaren; T Pradeep

    2006-09-01

    We report the synthesis of stearate functionalized nanobubbles of SiO2 with a few aniline molecules inside, represented as C6H5NH2@SiO2@stearate, exhibiting fluorescence with red-shifted emission. Stearic acid functionalization allows the materials to be handled just as free molecules, for dissolution, precipitation, storage etc. The methodology adopted involves adsorption of aniline on the surface of gold nanoparticles with subsequent growth of a silica shell through monolayers, followed by the selective removal of the metal core either using sodium cyanide or by a new reaction involving halocarbons. The material is stable and can be stored for extended periods without loss of fluorescence. Spectroscopic and voltammetric properties of the system were studied in order to understand the interaction of aniline with the shell as well as the monolayer, whilst transmission electron microscopy has been used to study the silica shell.

  7. Nepal CRS project incorporates.

    1983-01-01

    The Nepal Contraceptive Retail Sales (CRS) Project, 5 years after lauching product sales in June 1978, incorporated as a private, nonprofit company under Nepalese management. The transition was finalized in August 1983. The Company will work through a cooperative agreement with USAID/Kathmandu to complement the national family planning goals as the program continues to provide comtraceptives through retail channels at subsidized prices. Company objectives include: increase contraceptive sales by at least 15% per year; make CRS cost effective and move towards self sufficiency; and explore the possibility of marketing noncontraceptive health products to improve primary health care. After only5 years the program can point to some impressive successes. The number of retial shops selling family planning products increased from 100 in 1978 to over 8000, extending CRS product availability to 66 of the country's 75 districts. Retail sales have climbed dramatically in the 5-year period, from Rs 46,817 in 1978 to Rs 271,039 in 1982. Sales in terms of couple year protection CYP) have grown to 24,451 CYP(1982), a 36% increase over 1980 CYP. Since the beginning of the CRS marketing program, total distribution of contraceptives--through both CRS and the Family Planning Maternal and Child Haelth (FP/MCH) Project--has been increasing. While the FP/MCH program remains the largest distributor,contribution of CRS Products is increasing, indicating that CRS is creating new product acceptors. CRS market share in 1982 was 43% for condoms and 16% for oral contraceptives (OCs). CRS markets 5 products which are subsidized in order to be affordable to consumers as well as attractive to sellers. The initial products launched in June 1978 were Gulaf standard dose OCs and Dhaal lubricated colored condoms. A less expensive lubricates, plain Suki-Dhaal condom was introduced in June 1980 in an attempt to reach poorer rural populations, but rural distribution costs are excessive and Suki

  8. Incorporation and profile of fatty acids in tilapia fillets (Oreochromis niloticus fed with tung oil Incorporação e perfil de ácidos graxos em filés de tilápia (Oreochromis niloticus alimentada com óleo de tungue

    Elton Guntendorfer Bonafé

    2013-02-01

    Full Text Available The acceptance of tung oil enriched diet and the incorporation of conjugated linolenic acid - CLnA into fillets of Genetically Improved Farmed Tilapia (GIFT were investigated. The diet was well accepted, and after 10 days CLnA was incorporated into the fillets with a 1.02% content of total fatty acids (FA. In addition, biosynthesis of the conjugated linoleic acid isomers - CLA (0.31% of fillet total FA content from CLnA, and the presence of alpha-linolenic acid - LNA (1.08% of fillet total FA content, eicosapentaenoic acid - EPA (2.85% of fillet total FA content and docosahexaenoic acid - DHA (3.08% of fillet total FA content were observed. Therefore, the consumption of this fish can increase the intake of different FA (CLnA, CLA, LNA, EPA and DHA, which play an important role in human metabolism.A aceitação de tratamentos enriquecidos com óleo de tungue e a incorporação do ácido linolênico conjugado (CLnA em filés de tilápia melhorada geneticamente (GIFT foram investigadas. A dieta teve boa aceitação e, após 10 dias, o ClnA havia sido incorporado nos filés, mostrando um teor de 1,02% dos ácidos graxos totais (AG. Além disso, a biossíntese dos isômeros do ácido linoleico conjugado - CLA (0,31% de teor de AG total no filé, a partir do CLnA, e a presença do ácido alfalinolênico - LNA (1,08% de teor de AG total no filé, ácido eicosapentaenoico - EPA (2,85 % de teor de AG total no filé e ácido docosa-hexaenoico - DHA (3,08% de teor de AG total no filé foram observados. Assim, o consumo deste peixe pode aumentar a ingestão de diferentes AG (CLnA, CLA, LNA, EPA e DHA, que desempenham um papel importante no metabolismo humano.

  9. 14CO2 incorporation into C4 acids by leaves of the C3-C4 intermediate species, Moricandia arvensis and Panicum milioides, at Gamma-levels of CO2

    The crucifer M. arvensis is a species with photorespiratory characteristics intermediate between C3 and C4 plants. Although the precise mechanism promoting decreased photorespiration has yet to be elucidated, evidence to date disfavors participation of C4 metabolism. In order to conclusively assess the possible contribution, if any, of limited C4 metabolism to the reduction of photorespiration, detached leaves of M. arvensis and M. moricandioides (C3) were labeled with 14CO2. When M. arvensis was illuminated and pulsed for 10 s the amount of label in malate increased from 4.5% of total soluble dpm fixed at 21% O2 and 340 μl/l CO2 to 10.3% at 20 μl/l. A two-fold increase was also evident for aspartate (from 2.9% to 6.5%). The C3 Moricandia species failed to show a similar response to decreased CO2. The physiological significance of the increase in C4 acid-labeling which occurred in the intermediate species will be investigated by pulse-chase analyses of C4 acids and glycine. Similar data for Panicum milioides (C3-C4) will also be presented

  10. Thermogelling hydrogels of cyclodextrin/poloxamer polypseudorotaxanes as aqueous-based nail lacquers: application to the delivery of triamcinolone acetonide and ciclopirox olamine.

    Nogueiras-Nieto, Luis; Begoña Delgado-Charro, M; Otero-Espinar, Francisco J

    2013-04-01

    This work investigated the use of in situ gelling hydrogels based on polypseudorotaxanes of Pluronic F-127 and partially methylated β-cyclodextrin as aqueous nail lacquers. N-acetylcysteine and urea were incorporated as penetration enhancers. The formulations were tested for their ability to deliver ciclopirox and triamcinolone across human nail plate and bovine hoof. Simple aqueous solutions of the drugs with N-acetylcysteine provided measurable fluxes across hoof membranes but became quickly depleted of drug. Further, these solutions would have a short residence time upon nail application. Addition of Pluronic F-127 facilitated drug solubilization and provided the formulations with in situ gelling properties but drug entrapment into the micelles slowed down the delivery process. This was solved by addition of methylated β-cyclodextrin; the formulations retained the thermogelling properties, drug solubilization was further increased, and drug delivery was accelerated. The polymer chains compete with the drugs for the cyclodextrin cavity forming polypseudorotaxanes, which facilitated drug release. The permeability of both drugs was higher across bovine hoof than human nail. The new polypseudorotaxanes formulation delivered more ciclopirox across human nail than a marketed organic lacquer which supports the growing hypothesis that aqueous-based nail lacquers represent a superior formulation strategy in nail topical delivery. PMID:23201053