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Lysine
Kinds of Lysine Terms modified by Lysine Selected AbstractsVoltammetric Sensor for Sodium Nitroprusside Determination in Biological Fluids Using Films of Poly- L -LysineELECTROANALYSIS, Issue 9 2007Claudece Pereira, Francisco Abstract Sodium nitroprusside (NP), a commercial vasodilator, can be pre-concentrated on vitreous carbon electrode modified by films of 97.5%: 2.5% poly- L -lysine (PLL): glutaraldehyde (GA). This coating gives acceptable anion exchange properties whilst giving the required improvement of adhesion to the glassy carbon electrode surface. Linear response range and detection limit on nitroprusside in B-R buffer pH,4.0, were 1×10,6 to 2×10,5 mol L,1 and 1×10,7 mol L,1, respectively. The repeatability of the proposed sensor, evaluated in term of relative standard deviation, was measured as 4.1% for 10 experiments. The voltammetric sensor was directly applied to determination of nitroprusside in human plasma and urine samples and the average recovery for these samples was around 95,97% without any pre treatment. [source] Chemo-, Regio- and Stereospecific Synthesis of Unnatural, Fluorescent Amino Acids by Condensation of L -Lysine and 1-Vinylpyrrole-2-carbaldehydesEUROPEAN JOURNAL OF ORGANIC CHEMISTRY, Issue 24 2010Andrey V. Ivanov Abstract A new family of unnatural, optically active amino acids containing the pyrrole moiety have been synthesized by condensation of 1-vinylpyrrole-2-carbaldehydes with L -lysine under mild conditions (EtOH, room temp., 2.5,3 h, 0.5 wt.-% of CF3CO2H) in up to 90,% yields. Unlike non-vinylated analogues, 1-vinylpyrrole-2-carbaldehydes react chemo-, regio- and stereospecifically with an ,-amino group only to afford products of exclusively (E) configuration. The amino acids synthesized containing aromatic or condensed aromatic substitutents in the pyrrole ring fluoresce in the UV/Vis region (,max = 350,382 nm, Stokes shifts 6150,7800 cm,1). [source] New Low-Molecular-Mass Gelators Based on L -Lysine: Amphiphilic Gelators and Water-Soluble OrganogelatorsHELVETICA CHIMICA ACTA, Issue 1 2004Masahiro Suzuki The new L -lysine alkali-metal salts 1,5 (M+=Na+ and K+) with different alkyl groups at the N, -position were easily synthesized, and their hydro- and organogelation properties were investigated. All compounds were H2O-soluble, and some salts, especially the potassium salts, functioned as a hydrogenator that could gel water below 2 wt-%. These salts also had organogelation abilities for many organic solvents. [source] Effects of amino acid supplementation on the nutritive quality of fermented linseed meal protein in the diets for rohu, Labeo rohita, fingerlingsJOURNAL OF APPLIED ICHTHYOLOGY, Issue 5 2001N. Mukhopadhyay A feeding trial was conducted for 8 weeks to examine the effects of partial substitution of fish meal (FM) protein (crude protein content: 58.5%) with linseed meal protein with and without supplemental amino acids in diets for rohu Labeo rohita (Hamilton), fingerlings (mean weight: 1.50 ± 0.3 g). Prior to incorporation into the diets, linseed meal was fermented with lactic acid bacteria (Lactobacillus acidophilus) to reduce/eliminate the antinutritional tannin and phytic acid factors. Twelve experimental diets (diets D1,D12) were formulated to replace the FM protein from a reference diet (RD) with linseed meal protein at different levels (four sets of diets, of which each set of three diets contained 25%, 50% and 75% replacement of FM protein by linseed meal protein, respectively). Diets D1,D3 were not supplemented with any amino acid. Lysine was supplemented in diets D4,D6. Diets D7,D9 were supplemented with methionine + cystine (together), and diets D10,D12 contained lysine and methionine + cystine (together). Lysine and methionine + cystine (together) were added to the diets at 5.7% and 3.1% of dietary protein, respectively. The groups of fish fed diets without amino acid supplementation had significantly lower percentages of weight gain, specific growth rate and high feed : gain ratio than the fish groups fed other experimental diets. The addition of lysine and methionine + cystine to the diet in which 50% of the FM protein was replaced by linseed meal protein (diet D11) significantly improved fish performance. The results of the present study suggest that rohu fingerlings can effectively utilize the supplemented amino acids and that linseed meal protein can replace up to 50% of the FM protein in rohu diets if the linseed meal is properly processed (fermented) and supplemented with the lacking amino acids. [source] Lysyl Hydroxylase-2b Directs Collagen Cross-Linking Pathways in MC3T3-E1 Cells,JOURNAL OF BONE AND MINERAL RESEARCH, Issue 8 2004Suchaya Pornprasertsuk Abstract To elucidate the roles of LH2b in collagen cross-linking, MC3T3-E1 cell clones expressing higher (S) or lower (AS) levels of LH2b were established. Compared with controls, the collagen cross-linking pattern was shifted toward hydroxylysine-aldehyde (S clones)- or lysine-aldehyde (AS clones)-derived pathways. The data indicate that LH2b directs collagen cross-linking pathways through its action on telopeptidyl lysine residues. Introduction: Lysine (Lys) hydroxylation is a post-translational modification of collagen critical for cross-linking and glycosylation. Currently, three isoforms of lysyl hydroxylase (LH) have been identified, but their specific functions are still not well defined. Recently, we proposed that LH2 might modulate collagen cross-linking pattern through its action on Lys residues located in the telopeptide domains of collagen. Materials and Methods: To directly test this hypothesis, several MC3T3-E1 cell-derived clones expressing higher (sense [S]) or lower (antisense [AS]) levels of LH2b, the predominant form of LH2 in this cell line, were established and cultured for 2 weeks, and collagen cross-links and precursor aldehydes in the matrices were analyzed. Results: In S clones tested, the ratio of dihydroxylysinonorleucine (DHLNL) to hydroxylysinonorleucine (HLNL) was significantly higher than the average of controls (76% and 140% increase, respectively), and the level of pyridinoline (Pyr) was elevated (100% and 150% increase, respectively). In contrast, when MC3T3-E1 cells were transfected with a LH2b antisense construct (AS clones), the DHLNL/HLNL ratios were significantly lower than that of controls (56% and 73% decrease, respectively), and Pyr was not detected. Furthermore, significant amounts of an aldol-derived cross-link, dehydrohistidinohydroxymerodesmosine, were produced (,0.3 mol/mol of collagen) in AS clones. Conclusions: The data clearly show a critical role of LH2b in determining collagen cross-linking pathways, most likely through its action on telopeptidyl Lys residues. [source] Basic region of residues 228,231 of protein kinase CK1, is involved in its interaction with axin: Binding to axin does not affect the kinase activity,JOURNAL OF CELLULAR BIOCHEMISTRY, Issue 2 2005Pablo Sobrado Abstract Protein kinase CK1, also known as casein kinase 1, participates in the phosphorylation of ,-catenin, which regulates the functioning of the Wnt signaling cascade involved in embryogenesis and carcinogenesis. ,-catenin phosphorylation occurs in a multiprotein complex assembled on the scaffold protein axin. The interaction of CK1, from Danio rerio with mouse-axin has been studied using a pull-down assay that uses fragments of axin fused to glutathione S transferase, which is bound to glutathione sepharose beads. The results indicate that the three lysines present in the basic region of residues 228,231 of CK1, are necessary for the binding of CK1 to axin. Lysine 231 is particularly important in this interaction. In order to define the relevance of the axin-CK1, interaction, the effect of the presence of axin on the phosphorylating activity of CK1, was tested. It is also evident that the region of axin downstream of residues 503,562 is required for CK1, interaction. The binding of CK1, to axin fragment 292,681 does not facilitate the phosphorylation of ,-catenin despite the fact that this axin fragment can also bind ,-catenin. Binding of CK1, to axin is not required for the phosphorylation of axin itself and, likewise, axin does not affect the kinetic parameters of the CK1, towards casein or a specific peptide substrate. © 2004 Wiley-Liss, Inc. [source] Kinetics of Lysine and Other Amino Acids Loss During Extrusion Cooking of Maize GritsJOURNAL OF FOOD SCIENCE, Issue 2 2003S. Ilo ABSTRACT: Maize grits were extrusion-cooked in a conical, counter-rotating twin-screw extruder at different barrel temperatures, feed moistures, and screw speeds. Residence time distribution was measured by a dye tracer technique. Experiments with lysine-fortified maize grits showed a 1st order reaction for lysine loss. A detailed kinetic study has been performed for the losses during extrusion cooking of lysine, cystine, and arginine. The 1st-order rate constants were dependent mainly on product temperature and feed moisture, whereas screw speed had no influence. Activation energy of lysine, arginine, and cystine loss was 127, 68, and 76 kJ/mol, respectively. Shear stress significantly affected the rate constants of amino acids loss in extrusion cooking. [source] Effects of pH on Caramelization and Maillard Reaction Kinetics in Fructose-Lysine Model SystemsJOURNAL OF FOOD SCIENCE, Issue 7 2001E.H. Ajandouz ABSTRACT: The nonenzymatic browning reactions of fructose and fructose-lysine aqueous model systems were investigated at 100 °C between pH 4.0 and pH 12.0 by measuring the loss of reactants and monitoring the pattern of UV-absorbance and brown color development. At all the pH values tested, the loss of fructose was lower in the presence than in the absence of lysine. And, in lysine-containing fructose solution, the sugar disappeared more rapidly than the amino acid. Lysine was moderately lost below pH 8.0. Caramelization of fructose, which accounted for more than 40% of total UV-absorbance and 10 to 36% of brown color development, may therefore lead to overestimating the Maillard reaction in foods. [source] Immunolocalization of the High-Mobility Group N2 protein and acetylated histone H3K14 in early developing parthenogenetic bovine embryos derived from oocytes of high and low developmental competenceMOLECULAR REPRODUCTION & DEVELOPMENT, Issue 2 2008Guilherme M. Bastos Abstract This study investigated differences in the distribution of acetylated histone H3 at Lysine 14 (H3K14ac) and the High-Mobility Group N2 (HMGN2) protein in the chromatin of early- (before 24 hr) and late-cleaved (after 24 hr) bovine embryos derived from small- (1,2 mm) and large-follicles (4,8 mm). The presence of HMGN2 and H3K14ac has been associated with different nuclear functions including chromatin condensation, transcription, DNA replication and repair. In vitro matured oocytes were parthenogenetically activated (PA) and cultured in synthetic oviduct fluid medium. Early- and late-cleaved embryos were fixed at 36, 50, 60, 70 and 80 hr after PA to detect the presence of H3K14ac and HMGN2. The rates of nuclear maturation (81.1% vs. 58.7%), early cleavage (46.9% vs. 38.9%), and development to blastocyst stage (34.3% vs. 18.9%) were higher (P,<,0.05) in oocytes derived from large- compared to small follicles. The proportion of positively stained nuclei at 50 and 60 hr after PA was higher for both H3K14ac (27.2% vs. 4.8% and 64.3% vs. 30%) and HMGN2 (47% vs. 21.3% and 60.6% vs. 46%) in early versus late cleaved embryos derived from small- versus large-follicles, respectively. However, the rate of positive nuclei in early-cleaved embryos from small-versus large-follicles was similar for HMGN2 (87% vs. 93%) but lower for H3K14ac (51% vs. 64.4%) at 80 hr after PA. These data suggest that less developmentally competent embryos derived from small follicles had an altered chromatin remodeling process at the early stages of development compared to those derived from large follicles that are more competent to support development to blastocyst stage. Mol. Reprod. Dev. 75: 282,290, 2008. © 2007 Wiley-Liss, Inc. [source] Magnetic resonance imaging and biological properties of pancreatic islets labeled with iron oxide nanoparticlesNMR IN BIOMEDICINE, Issue 8 2009Hoe Suk Kim Abstract This study was undertaken to investigate the in vitro effect of islet labeling with iron oxide nanoparticles for MRI on islet viability, insulin secretion, and gene expression. Isolated rat islets were labeled with Resovist (25,200,µg Fe/mL, a clinically approved MRI contrast agent) in the presence or absence of poly- l -Lysine (PLL, 1.5,µg/mL) for 48,h. The iron content of labeled islets was found to increase in a dose-dependent manner. More than 90% of the islets were labeled with 100,µg Fe/mL. We confirmed the localizations of iron oxide nanoparticles within islet , -cells by insulin immunostaining. As the concentration of Resovist increased, T2 values as determined by T2 -weighted MRI on a 1.5,Tesla MR scanner decreased. Labeling of 100 islets in a medium containing 100,µg Fe/mL of Resovist in the absence of PLL provided sufficient contrast for islet visualization on T2 -weighted MRI. MTT assays showed that the viability of labeled islets was not different from that of unlabeled islets. No statistical difference was observed between labeled (2.91,±,0.36) and unlabeled islets (2.83,±,0.61) in terms of the ability to secrete insulin, as determined by the glucose stimulation index. We also evaluated the effect of iron oxide incorporation on the gene expressions in islet cells using RT-PCR (reverse transcriptase PCR). Insulin expression in labeled islets was significantly elevated (1.83,±,0.25 fold vs. unlabeled; p,=,0.005), but not the expression of somatostatin (1.39,±,0.18 fold vs. unlabeled; p,=,0.085) or glucagons (1.28,±,0.13 fold vs. unlabeled; p,=,0.09). Expression of an important transcription factor for insulin gene transcription, BETA2 (beta-cell E-box trans-activator), was increased in labeled islets (1.67,±,0.15 fold vs. unlabeled; p,=,0.029). The findings of this study indicate that Resovist provides a satisfactory means to image islets and has no deleterious effect on islet function or gene expression. Copyright © 2009 John Wiley & Sons, Ltd. [source] Effect of dietary lysine and methionine supplementation on growth, nutrient utilization, carcass compositions and haemato-biochemical status in Indian Major Carp, Rohu (Labeo rohita H.) fed soy protein-based dietAQUACULTURE NUTRITION, Issue 4 2009P. SARDAR Abstract An 8-week feeding trial was conducted in flow through system to examine the effects of dietary supplementation of lysine and methionine on growth, nutrient utilization, haemato-biochemical status and carcass compositions in Indian major carp, rohu, Labeo rohita fingerlings (average weight 6.32 ± 0.06 g). Four experimental soy protein-based diets D0 (without lysine or methionine supplementation), D1 (lysine supplementation alone), D2 (methionine supplementation alone) and D3 (both lysine and methionine supplementation) were fed to triplicate groups. l -Lysine and dl -methionine were added to the diets containing 550 g kg,1 soybean meals at 4 and 7 g kg,1 of dry diet respectively. Significant higher weight gain, specific growth rate (SGR), protein efficiency ratio (PER), dry matter retention, nitrogen retention, total ash retention, whole carcass protein, haemoglobin concentration, haematocrit value, total erythrocytic count, total leucocytic count, plasma glucose and plasma total protein and lower FCR, per cent lipid retention and whole body moisture content were observed in fish fed soya protein-based diet supplemented with both lysine and methionine than that of fish of other dietary groups at the end of 8 weeks feeding trial. Although fish fed diet supplemented with either methionine or lysine did not show any significant differences of growth performances, feed utilization, carcass composition and haemato-biochemical status, fish of both of these dietary groups showed significantly better growth performances, feed utilization, carcass composition and haemato-biochemical status than that of fish fed diet without lysine and methionine supplementation. [source] Partial or total replacement of fishmeal by solvent-extracted cottonseed meal in diets for juvenile rainbow trout (Oncorhynchus mykiss)AQUACULTURE NUTRITION, Issue 6 2006L. LUO Abstract The effect of solvent-extracted cottonseed meal (SCSM) as a partial or total replacement of fishmeal was studied in juvenile rainbow trout (Oncorhynchus mykiss). Six experimental diets SCSM0, SCSM25, SCSM50, SCSM75, SCSM75A and SCSMT, containing a gradient of SCSM 0, 152, 305, 465, 460 and 610 g kg,1 to replace 0, 112.5, 225, 337.5, 337.5 and 450 g kg,1 fishmeal protein were fed to triplicate groups (initial body weight of 39.2 ± 0.1 g) for 8 weeks. The diet SCSM75A was supplemented with lysine and methionine, to be similar to SCSM0 for juvenile rainbow trout. Faeces were colleted after 4 weeks of normal feeding for apparent digestibility coefficients (ADC) of dry matter, crude protein and gross energy determination. Total replacement of fishmeal adversely affected growth performance. Fish fed with diet SCSMT had significantly (P < 0.05) lower weight gain, specific growth ratio, feed conversion efficiency (FCE) and protein efficiency ratio than fish fed with other diets. The FCE of SCSM75 and SCSM75A were significantly lower (P < 0.05) than those of fish fed with SCSM0 diets. The ADC of the dry matter of SCSM75 and SCSMT were significantly lower than the SCSM0 diet, and the ADC of crude protein and the energy of SCSMT were the lowest (P < 0.05). The ADC of threonine, proline, alanine, valine, isoleucine, leucine, lysine and methionine of fish fed with diet SCSMT were lower. Lysine and methionine supplement positively affected the ADC of SCS75A diet. There were no significant differences in the fish body composition. It is shown that SCSM can be utilized in the juvenile rainbow trout diet up to 305 g kg,1, to replace about 50% of fishmeal protein in this experiment. [source] Corrigendum: A Genetically Encoded ,- N -Methyl Lysine in Mammalian CellsCHEMBIOCHEM, Issue 8 2010Dan Groff Dr. No abstract is available for this article. [source] Synthesis and Proinflammatory Properties of Muramyl Tripeptides Containing Lysine and Diaminopimelic Acid MoietiesCHEMBIOCHEM, Issue 11 2005Abhijit Roychowdhury Dr. Abstract The unusual amino acid diaminopimelic acid (DAP) was prepared by cross metathesis of appropriately protected vinyl glycine and allyl glycine derivatives. Catalytic hydrogenation of the cross-coupling product resulted in reduction of the double bond and the removal of protecting groups. The resulting compounds were appropriately protected for the polymer-supported and solution-phase synthesis of muramyl tripeptides 2 and 3, which differ in the amidation of the ,-carboxylic acids of the isoglutamine and DAP moieties. Muramyl dipeptide (1, MDP), the DAP-containing muramyl tripeptide 3, and the lysine-containing muramyl tripeptides 4 and 5 induced TNF-, gene expression without TNF-, protein production in a human monocytic cell line. The observed block in translation could be removed by co-incubation with LPS, resulting in an apparent synergistic effect. Compound 2 did not induce TNF-, gene expression, neither did it exhibit a synergistic effect with LPS; this indicates that amidation of the ,-carboxylic acids of the isoglutamine and DAP moieties results in a loss of biological activity. It is proposed that amidation of ,-carboxylic acids is a strategy that may be used by pathogens to avoid detection by the innate immune system. Furthermore, the pattern recognition receptors Nod1 and Nod2 have been implicated in the possible induction of a synergistic effect of muropeptides with LPS. [source] ChemInform Abstract: Protonated Arginine and Lysine as Catalysts for the Direct Asymmetric Aldol Reaction in Ionic Liquids.CHEMINFORM, Issue 10 2009Marco Lombardo Abstract ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 200 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a "Full Text" option. The original article is trackable via the "References" option. [source] A Convenient and Efficient Synthesis of (S)-Lysine and (S)-Arginine Homologues via Olefin Cross-Metathesis.CHEMINFORM, Issue 45 2005Timothy P. Boyle No abstract is available for this article. [source] ChemInform Abstract: A New Receptor Molecule for Lysine and Histidine in Water: Strong Binding of Basic Amino Acid Esters by a Macrocyclic Host.CHEMINFORM, Issue 37 2001Thomas Grawe Abstract ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 100 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a "Full Text" option. The original article is trackable via the "References" option. [source] Virtual Screening and Biological Characterization of Novel Histone Arginine Methyltransferase PRMT1 InhibitorsCHEMMEDCHEM, Issue 1 2009Ralf Heinke Abstract Lysine and arginine methyltransferases participate in the posttranslational modification of histones and regulate key cellular functions. Protein arginine methyltransferase,1 (PRMT1) has been identified as an essential component of mixed lineage leukemia (MLL) oncogenic complexes, revealing its potential as a novel therapeutic target in human cancer. The first potent arginine methyltransferase inhibitors were recently discovered by random- and target-based screening approaches. Herein we report virtual and biological screening for novel inhibitors of PRMT1. Structure-based virtual screening (VS) of the Chembridge database composed of 328,000 molecules was performed with a combination of ligand- and target-based in,silico approaches. Nine inhibitors were identified from the top-scored docking solutions; these were experimentally tested using human PRMT1 and an antibody-based assay with a time-resolved fluorescence readout. Among several aromatic amines, an aliphatic amine and an amide were also found to be active in the micromolar range. [source] Lysine-based peptide nucleic acids (PNAs) with strong chiral constraint: Control of helix handedness and DNA binding by chiralityCHIRALITY, Issue S1 2005Tullia Tedeschi Abstract Two enantiomeric chiral PNAs bearing three adjacent d - or l -lysine-based residues in the middle of the strand ("chiral box" PNAs, sequence H-GTAGALysTLysCLysACT-NH2) have been used as models in order to comprehensively study the effects of the stereogenic centers on PNA conformation and on PNA binding properties to complementary PNA and DNA strands. The binding properties of the two enantiomeric PNAs and of their homologous achiral PNA have been extensively studied by UV and CD spectroscopy and by mass spectrometry, both in the antiparallel and in the parallel mode with complementary PNA and DNA strands. In the antiparallel PNA:PNA duplexes, l -Lys PNA were found to form left-handed, and d -Lys PNA right handed helices, while in parallel duplexes, the reversed helicities were observed. Correspondingly, the preferred mode of binding and the best mismatch recognition of the d -Lys containing PNA with (right handed) DNA was found to be in the antiparallel orientation, while that of l -Lys PNA was found to be in the parallel mode. A rationale which correlates the preferred handedness of the PNA-PNA duplexes to the directionality of the binding to complementary DNA duplexes has been devised according to structural data and considering the "retro,inverso" concept widely used for peptides. Chirality 17:S196,S204, 2005. © 2005 Wiley-Liss, Inc. [source] Chapter 6: Maize with Increased Lysine (Lysine Maize,LY038)COMPREHENSIVE REVIEWS IN FOOD SCIENCE AND FOOD SAFETY, Issue 1 2008Article first published online: 30 JAN 200 ABSTRACT:, Data and information provided in this case study relate to a crop derived by modern biotechnology, in which a specific nutrient (lysine) has been increased in maize grain.Lysine maize is a feed ingredient with enhanced nutritional characteristics for poultry and swine and provides an alternative to adding supplemental lysine to diets for these animals. Lysine maize is in an advanced state of development; therefore, extensive unpublished data and information are presented to demonstrate that (1) Lysine maize,and the feeds and foods derived from it,are as safe as those derived from conventional maize,and (2) the increased lysine in Lysine maize grain produces the intended nutritional benefit for broiler chickens when compared to a diet containing conventional maize grain and a crystalline lysine supplement. These conclusions are based on a detailed molecular characterization of Lysine maize,a safety assessment of the introduced protein,a safety and nutritional assessment of the LY038 crop,and a comparison of the agronomic and phenotypic properties of maize hybrids with and without the Lysine maize trait. Although Lysine maize is a specialty crop for use in animal feed,its safety for both animals and humans must be demonstrated. Free lysine is significantly increased in Lysine maize by the introduction of the dapA gene (cordapA) from Corynebacterium glutamicum that encodes a form of dihydrodipicolinate synthase (cDHDPS) that is insensitive to lysine feedback inhibition.Analysis of lysine anabolic and catabolic pathways in maize identified 6 metabolites that might change as a consequence of the introduction of cDHDPS insensitive to lysine-feedback inhibition. The results of compositional analysis demonstrated that Lysine maize grain is comparable to conventional maize, with the exception of the intended increase in lysine and a corresponding increase in 2 products of lysine catabolism,saccha-ropine and -aminoadipic acid. Therefore, the safety and/or nutritional implication of these 3 compounds under the conditions of use were the focus of additional assessments and found to not present either a safety or nutritional problem. [source] Use of quasi-isoelectric buffers as anolyte and catholyte to improve capillary isoelectric focusing performancesELECTROPHORESIS, Issue 8 2008Martine Poitevin Abstract The use of quasi-isoelectric anolytes and catholytes has been investigated to improve CIEF performances. Narrow pH cuts of carrier ampholytes (NC) have been compared to more conventional couples of anolytes/catholytes (phosphoric acid/sodium hydroxide and glutamic acid/lysine). First, a CIEF setup that consists in a bare silica capillary and 70:30 water/glycerol separation medium has been used. The experiments have shown that when using NC instead of more classical anolytes and catholytes, an increase in the protein detection time was observed and the resolutions obtained for neutral and acidic proteins were doubled. Moreover, according to the NC fraction used, the resolution was modified. In order to investigate further the mechanisms involved, a second setup using a capillary coated with hydroxypropylcellulose was used. With this setup no difference has been observed when changing anolyte and catholyte nature. A simple methodology has then been developed to evaluate EOF during focusing and mobilization steps of CIEF experiments. It highlighted the crucial role played by EOF when using a bare silica capillary. EOF indeed decreased by 33% during mobilization step when using NC instead of classical anolytes and catholytes. [source] Changes in calcium absorption and subsequent tissue distribution induced by Maillard reaction products: in vitro and in vivo assays,JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 2 2006Cristina Delgado-Andrade Abstract The effects of Maillard reaction products (MRP) from glucose,lysine and glucose,methionine on calcium bioavailability were studied by in vivo (rats) and in vitro (Caco-2 cells) assays. Equimolar glucose/lysine and glucose/methionine mixtures (40% moisture) were heated (150 °C, 30 min) to prepare samples (GL30 and GM30, respectively). For 21 days, rats were fed a control diet (control group) or diets containing separately 3% of the heated mixtures (GL30 and GM30 groups, respectively). In the last week a calcium balance was performed, after which the animals were sacrificed and some organs and serum were removed to analyze calcium levels. A second balance was carried out throughout the experimental period to calculate global calcium retention (retained calcium during the entire 21 days). Unheated and heated samples were used for calcium transport experiments in Caco-2 cells. Food intake and final body weight were lower in the GM30 group. Calcium fecal excretion decreased and digestibility increased in this group. Accordingly, increased calcium transport in Caco-2 cells was found in the presence of the GM30 sample, when compared with the unheated sample. However, global calcium retention tended to decrease in the GM30 group, mainly owing to the lower food intake. Bone calcium concentrations decreased in the animals fed the MRP diets. The possible long-term effects of MRP intake on calcium digestibility and bone calcium should be taken into account to avoid related diseases. Copyright © 2005 Society of Chemical Industry [source] MR imaging for the longevity of mesenchymal stem cells labeled with poly- L -lysine,Resovist complexesCONTRAST MEDIA & MOLECULAR IMAGING, Issue 2 2010Gang Liu Abstract Superparamagnetic iron oxide (SPIO) nanoparticles are emerging as ideal probes for noninvasive cell tracking. In this study, poly- L -lysine (PLL) was mixed with Resovist to form the PLL,Resovist complexes and the control of the complexes formed by PLL and Resovist and their subsequent properties was easily achievable. MSCs could be safely and efficiently labeled for MR imaging using PLL,Resovist complexes (w/w 0.01:1) and the labeled MSCs could be detected to have definite decreased signal intensity on T2 -weight imaging until 20 days with standard 1.5,T MR equipment. This study describes a simple protocol to label MSCs using PLL,Resovist complexes and the results presented in our study can provide a basis for the application of PLL,Resovist complexes cell labeling. Copyright © 2010 John Wiley & Sons, Ltd. [source] Molecular recognition of sugars by lanthanide (III) complexes of a conjugate of N, N -bis[2-[bis[2-(1, 1-dimethylethoxy)-2-oxoethyl]amino]ethyl]glycine and phenylboronic acidCONTRAST MEDIA & MOLECULAR IMAGING, Issue 4 2007Elisa Battistini Abstract A novel conjugate of phenylboronic acid and an Ln(DTPA) derivative, in which the central acetate pendant arm was replaced by the methylamide of L -lysine, was synthesized and characterized. The results of a fit of variable 17O NMR data and a 1H NMRD profile show that the water residence lifetime of the Gd(III) complex (150,ns) is shorter than that of the parent compound Gd(DTPA)2, (303,ns). Furthermore, the data suggest that several water molecules in the second coordination sphere of Gd(III) contribute to the relaxivity of the conjugate. The Ln(III) complexes of this conjugate are highly suitable for molecular recognition of sugars. The interaction with various sugars was investigated by 11B NMR spectroscopy. Thanks to the thiourea function that links the phenylboronic acid targeting vector with the DTPA derivative, the interactions are stronger than that of phenylboronic acid itself. In particular, the interaction with N -propylfructosamine, a model for the glucose residue in glycated human serum albumin (HSA), is very strong. Unfortunately, the complex also shows a rather strong interaction with hexose-free HSA (KA,=,705,±,300). Copyright © 2007 John Wiley & Sons, Ltd. [source] Regulation of oocyte maturation in fishDEVELOPMENT GROWTH & DIFFERENTIATION, Issue 2008Yoshitaka Nagahama A period of oocyte growth is followed by a process called oocyte maturation (the resumption of meiosis) which occurs prior to ovulation and is a prerequisite for successful fertilization. Our studies using fish models have revealed that oocyte maturation is a three-step induction process involving gonadotropin (LH), maturation-inducing hormone (MIH), and maturation-promoting factor (MPF). LH acts on the ovarian follicle layer to produce MIH (17,, 20,-dihydroxy-4-pregnen-3-one, 17,, 20,-DP, in most fishes). The interaction of ovarian thecal and granulosa cell layers (two-cell type model), is required for the synthesis of 17,,20,-DP. The dramatic increase in the capacity of postvitellogenic follicles to produce 17,,20,-DP in response to LH is correlated with decreases in P450c17 (P450c17-I) and P450 aromatase (oP450arom) mRNA and increases in the novel form of P450c17 (P450c17-II) and 20,-hydroxysteroid dehydrogenase (20,-HSD) mRNA. Transcription factors such as Ad4BP/SF-1, Foxl2, and CREB may be involved in the regulation of expression of these steroidogenic enzymes. A distinct family of G-protein-coupled membrane-bound MIH receptors has been shown to mediate non-genomic actions of 17,, 20,-DP. The MIH signal induces the de novo synthesis of cyclin B from the stored mRNA, which activates a preexisting 35 kDa cdc2 kinase via phosphorylation of its threonine 161 by cyclin-dependent kinase activating kinase, thus producing the 34 kDa active cdc2 (active MPF). Upon egg activation, MPF is inactivated by degradation of cyclin B. This process is initiated by the 26S proteasome through the first cut in its NH2 terminus at lysine 57. [source] Epigenetic control of translation regulation: Alterations in histone H3 lysine 9 post-translation modifications are correlated with the expression of the translation initiation factor 2B (Eif2b5) during thermal control establishmentDEVELOPMENTAL NEUROBIOLOGY, Issue 2 2010Tatiana Kisliouk Abstract Thermal control set point is regulated by thermosensitive neurons of the preoptic anterior hypothalamus (PO/AH) and completes its development during postnatal critical sensory period. External stimuli, like increase in environmental temperature, influence the neuronal protein repertoire and, ultimately, cell properties via activation or silencing of gene transcription, both of which are regulated by the "histone code."" Here, we demonstrated an increase in global histone H3 lysine 9 (H3K9) acetylation as well as H3K9 dimethylation in chick PO/AH during heat conditioning at the critical period of sensory development. In contrast to the global profile of H3K9 modifications, acetylation and dimethylation patterns of H3K9 at the promoter of the catalytic subunit of eukaryotic translation initiation factor 2B (Eif2b5) were opposite to each other. During heat conditioning, there was an increase in H3K9 acetylation at the Eif2b5 promoter, simultaneously with decrease in H3K9 dimethylation. These alterations coincided with Eif2b5 mRNA induction. Moreover, exposure to excessive heat during the critical period resulted in long-term effect on both H3K9 tagging at the Eif2b5 promoter and Eif2b5 mRNA expression. These data suggest a role for dynamic H3K9 post-translational modifications in global translation regulation during the thermal control establishment. © 2009 Wiley Periodicals, Inc. Develop Neurobiol, 2010 [source] Laminin and fibronectin modulate inner ear spiral ganglion neurite outgrowth in an in vitro alternate choice assayDEVELOPMENTAL NEUROBIOLOGY, Issue 13 2007Amaretta R. Evans Abstract Extracellular matrix (ECM) molecules have been shown to function as cues for neurite guidance in various populations of neurons. Here we show that laminin (LN) and fibronectin (FN) presented in stripe micro-patterns can provide guidance cues to neonatal (P5) inner ear spiral ganglion (SG) neurites. The response to both ECM molecules was dose-dependent. In a LN versus poly- L -lysine (PLL) assay, neurites were more often observed on PLL at low coating concentrations (5 and 10 ,g/mL), while they were more often on LN at a high concentration (80 ,g/mL). In a FN versus PLL assay, neurites were more often on PLL than on FN stripes at high coating concentrations (40 and 80 ,g/mL). In a direct competition between LN and FN, neurites were observed on LN significantly more often than on FN at both 10 and 40 ,g/mL. The data suggest a preference by SG neurites for LN at high concentrations, as well as avoidance of both LN at low and FN at high concentrations. The results also support a potential model for neurite guidance in the developing inner ear in vivo. LN, in the SG and osseus spiral lamina may promote SG dendrite growth toward the organ of Corti. Within the organ of Corti, lower concentrations of LN may slow neurite growth, with FN beneath each row of hair cells providing a stop or avoidance signal. This could allow growth cone filopodia increased time to sample their cellular targets, or direct the fibers upward toward the hair cells. © 2007 Wiley Periodicals, Inc. Develop Neurobiol, 2007 [source] Electrocatalytic Reduction and Determination of Iodate and Periodate at Silicomolybdate-Incorporated-Glutaraldehyde- Cross-Linked Poly- L -lysine Film ElectrodesELECTROANALYSIS, Issue 10 2010Yu-Ching Pan Abstract The present work describes reduction of iodate (IO3,), and periodate (IO4,) at silicomolybdate-doped-glutaraldehyde-cross-linked poly- L -lysine (PLL-GA-SiMo) film coated glassy carbon electrode in 0.1,M H2SO4. In our previous study, we were able to prepare the PLL-GA-SiMo film modified electrode by means of electrostatically trapping SiMo12O404, mediator in the cationic film of PLL-GA, and the voltammetric investigation in pure supporting indicated that the charge transport through the film was fast. Here, the electrocatalytic activity of PLL-GA-SiMo film electrode towards iodate and periodate was tested and subsequently used for analytical determination of these analytes by amperometry. The two electron reduced species of SiMo12O404, anion was responsible for the electrocatalytic reduction of IO3, at PLL-GA-SiMo film electrode while two and six electron reduced species were showed electrocatalytic activity towards IO4, reduction. Under optimized experimental conditions of amperometry, the linear concentration range and sensitivity are 2.5×10,6 to 1.1×10,2,M and 18.47,,A mM,1 for iodate, and 5×10,6 to 1.43×10,4,M and 1014.7,,A mM,1 for periodate, respectively. [source] Construction of L -Lysine Sensor by Layer-by-Layer Adsorption of L -Lysine 6-Dehydrogenase and Ferrocene-Labeled High Molecular Weight Coenzyme Derivative on Gold ElectrodeELECTROANALYSIS, Issue 24 2008Haitao Zheng Abstract A ferrocene-labeled high molecular weight coenzyme derivative (PEI-Fc-NAD) and a thermostable NAD-dependent L -lysine 6-dehydrogenase (LysDH) from thermophile Geobacillus stearothermophilus were used to fabricate a reagentless L -lysine sensor. Both LysDH and PEI-Fc-NAD were immobilized on the surface of a gold electrode by consecutive layer-by-layer adsorption (LBL) technique. By the simple LBL method, the reagentless L -lysine sensor, with co-immobilization of the mediator, coenzyme, and enzyme was obtained, which exhibited current response to L -lysine without the addition of native coenzyme to the analysis system. The amperometric response of the sensor was dependent on the applied potential, bilayer number of PEI-Fc-NAD/LysDH, and substrate concentration. A linear current response, proportional to L -lysine concentration in the range of 1,120,mM was observed. The response of the sensor to L -lysine was decreased by 30% from the original activity after one month storage. [source] Comparison of Electrochemical and Surface Plasmon Resonance Immunosensor Responses on Single Thin FilmELECTROANALYSIS, Issue 20 2008Ryoji Kurita Abstract This paper reports results obtained when comparing an electrochemical enzyme immunosensor and a surface plasmon resonance (SPR) based immunosensor on the same gold surface installed in an electrochemical SPR flow cell. Simultaneous electrochemical and SPR measurements were performed on a gold surface modified with multilayers of poly- L -lysine and poly-styrenesulfonate assembled with the layer-by-layer method. First, we obtained the SPR response induced by the formation of an immunocomplex from the shift in the SPR angle by injecting an anti tumor necrosis factor-, antibody solution labeled with alkaline phosphatase into the flow cell containing the multilayer modified with tumor necrosis factor-,. Then we compared this SPR result with that obtained for the electrochemical oxidation current of p -aminophenol catalyzed by alkaline phosphatase from p -aminophenolphosphate on the same gold film. We compared the two immunosensor responses obtained using the different measurement principles and found that there was a high correlation efficient of 0.973 between them. This was because we were able to immobilize the immunoreagents with good stability and without losing the transport of the enzyme product in the multilayer whose thickness we easily controlled with nanometer scale accuracy. We also report that the detection limit of our electrochemical immunosensor after optimization was around 100,pg/mL (0.4,pM), which is one of the lowest values yet reported for an electrochemical immunosensor. [source] | ||||||||||||||||||||||||||||||||||||||||