children and youth aged 3 to 21 years

... 2 73. 15 Kelvins (K) 8 .31 4471 J K21 mol21 (+15)a 1.987 ca1th K21 mol21 0.0820575 dm3 atm K21 mol21 6.0 2215 Â10 23 mol21 (+107)a 96485 .33 83 C mol21 (+ 839 )a 1.6 0217 6 53 1 0219 C (+1 43) a 1 .38 06505Â102 23 ... Solution and an Ideal Liquid Solution / 33 2 Composition of the Two Phases in Equilibrium / 33 2 Temperature Dependence of the Equilibrium Compositions / 33 3 Exercises / 33 3 References / 33 5 15 DILUTE ... of Activities and Activity Coefficients / 35 8 Activity / 35 8 Activity Coefficient / 35 8 16.2 Choice of Standard States / 35 9 Gases / 35 9 Liquids and Solids / 36 0 16 .3 Gibbs Function and the Equilibrium...

Ngày tải lên: 02/04/2014, 16:43

... to be 121 124 129 129 130 130 131 131 132 132 132 133 133 134 134 134 134 135 135 136 136 136 136 137 137 138 138 138 140 1 43 Create a histogram with bins extending from 120–125, 125– 130 , and so ... 2.76 0 .33 0.81 1.12 1.62 3. 05 0 .36 0.82 1.12 1.66 0 .38 0.87 1. 13 1.68 0.46 0.87 1.20 1.68 0.47 0.87 1.20 1.70 0.60 0.91 1 .32 1.78 0.61 0.96 1 .33 1.82 0.61 0.97 1 .35 1.89 0.66 0.98 1 .38 1. 93 0.67 ... 28 For the observations 21, 36 , 42, 24, 25, 36 , 35 , 49, 32 ¯ verify that the sample mean, trimmed mean and median are X = 33 .33 , ¯ t = 32 .9 and M = 35 X 29 The largest observation in the last...

Ngày tải lên: 11/06/2014, 00:51

... Equipment and materials 3. 2.2 Methodology……… 3. 2 .3 Simulation… 3. 3 Results and discussion 3. 3.1 Experimental 3. 3.2 Simulation 3. 4 Conclusions ... Simulation; Empty symbol: Experimental)…… 85 3. 14 3. 15 3. 16 3. 17 3. 18 3. 19 3. 20 xvi 3. 21 3. 22 4.1 4.2 4 .3 4.4 4.5 4.6 4.7 4.8 4.9 4.10 4.11 Simulated and experimental flow pattern in TME (Viewed ... 91 92 92 93 93 97 4 .3 Results and discussion 4 .3. 1 SU-8 mater fabrication………………… 4 .3. 2 PDMS replicates (daughter mold)…………… 4 .3. 3 Sacrificial template………………… 4 .3. 4 Hot embossing…………………………...

Ngày tải lên: 14/11/2014, 13:26

... buffer to the column through an injector Monitor the eluate on-line with an ultraviolet (UV) monitor and collect fractions using a fraction collector and subject to the appropriate carbohydrate and ... running buffer to the column through an injector Monitor the eluate on-line with a UV monitor and collect fractions using a fraction collector and subject to the appropriate carbohydrate and antibody ... and apply the sample to the column Run the column in a linear gradient up to 100% buffer B or D Monitor the eluate on-line with a UV monitor and collect fractions using a fraction collector and...

Ngày tải lên: 08/10/2012, 10:25

... fuchsin-alcian blue staining Am J Clin Pathol 33 , 4 53 13 Spicer, S S (1965) Diamine methods for differentiating mucosubstances histochemically J Histochem Cytochem 13, 211 – 234 14 Williams, G T (1985) Transitional ... mM Add lectin to a final concentration one-fifth that of the inhibiting sugar and incubate for 30 to h Proceed with histochemistry protocol as usual 3. 2 .3 Enzymatic Deglycosylation to Confirm Lectin ... Subheadings 3. 6.1. 3. 6.4 Histological Methods for Detection of Mucin 37 3. 3 Mucin Immunohistochemistry (see Notes 17–19): Standard ABC Immunoperoxidase Technique for Paraffin Sections 10 11 12 13 14...

Ngày tải lên: 08/10/2012, 10:25

... metabisulfite and dry the membrane in a warm airstream 3. 2 .3 Detection of Mucins on Membranes Using Antibodies or Lectins (see Notes 10,12 and 13) 3. 2 .3. 1 BLOCKING Membranes need to be blocked ... Humana, Totowa, NJ Carraway, K L (2000) Preparation of membrane mucin, in Glycoprotein Methods and Protocols: The Mucins (Corfield, T ed.), Humana, Totowa, NJ Nagma, K., Thornton, D J., Khan, N., and ... The use of wheat germ agglutinin to improve binding of heterogeneous mucin species to nitrocellulose membranes Anal Biochem 219 , 37 3 37 5 21 Jay, G D., Culp, D J., and Jahnke, M R (1990) Silver staining...

Ngày tải lên: 08/10/2012, 10:25

... Williams, G T., Allen, A., and Rhodes, J (1994) Thickness of adherent mucus gel on colonic mucosa in humans and its relevance to colitis Gut 35 , 35 3 35 9 28 Sandzen, B., Blom, H., and Dahlgren, S (1988) ... 174–179 Bolton, W P., Palmer, D., and Cohen, M M (1978) Stimulation of mucus and nonparietal cell secretion by the E2 prostaglandins Digest Dis Sci 23, 35 9 36 4 McQueen, S., Allen, A., and Garner, ... Sci 31 , 133 8– 134 4 64 Allen and Pearson 13 Morris, G P., Harding, R K., and Wallace, J L (1984) A functional model for extracellular gastric mucus in the rat Virchows Arch [Cell Pathol.] 46, 239 –251...

Ngày tải lên: 08/10/2012, 10:25

... extensive protocol for quantitation of the biosynthesis and secretion of mucin, which is discussed in Subheadings 3. 2 and 3. 3 Measure protein concentrations according to the BCA protein assay and calculate ... [35 S]methionine/ [35 S]cysteine b [35 S]sulfate 17 Water bath, 37 °C 18 Whatman 3MM filter paper 19 Molecular weight marker: nonreduced monomeric and dimeric rat gastric mucin precursors, molecular mass 30 0 and 600 kDa, ... apparatus and air-dry for Perform all ensuing procedures in Blotto as follows: a Incubate in Blotto for 30 min, and incubate with antibody directed against nonglycosylated peptide epitopes of...

Ngày tải lên: 08/10/2012, 10:25

... buffer to each sample and vortex to mix well Leave at room temperature for 30 Remove excess PITC by centrifugal evaporation under vacuum for h The derivatized samples can be stored at –20°C 3. 4 .3. 3 ... respiratory, cervical, and salivary secretions, as well as from respiratory cells in culture, and have identified a pattern of four major peptides (masses 1 036 , 1 132 , 1688, and 1980 Daltons; some ... peptides by automated N-terminal sequencing Use sequence to search sequence databases 3. 4 .3 Amino Acid Analysis (see Note 9) 3. 4 .3. 1 ACID HYDROLYSIS Dissolve the samples (1 µg to mg) in 500 µL...

Ngày tải lên: 08/10/2012, 10:25

... positively stained, see Subheading 3. 3.1 .3. ; if not, go to the next step Wash grid in 95% ethanol, remove excess solution on filter paper, and air-dry 3. 3.1 .3 STAINING AND SHADOWING Dip grid in staining ... 36 1 37 7 11 Lang D and Mitani, M (1970) Simplified quantitative electron microscopy of biopolymers Biopolymers 9, 37 3 37 9 96 Sheehan and Thornton 12 Koller, T., Harford, A G., Lee, Y K., and Beer, M ... poured into a Teflon trough A drop (1 µL) of a solution of BAC (100 µg/mL) is touched to the surface and the solution is left for 5–15 In this time the mucins diffuse to the surface and become...

Ngày tải lên: 08/10/2012, 10:25

... Lys Total Standard (area)a Bovine conjunctival mucin (area) 258,795,760 215 ,720,688 249,767,456 209,927 ,39 2 1 43, 969,008 2 13, 371,584 2 03, 866,048 2 13, 5 53, 008 216 ,618,864 91, 930 ,856 200,286 ,32 0 2 03, 002,480 ... 7, 537 ,291 84,959,272 169,595,904 29,927 ,38 0 132 , 131 ,184 270,782,624 104,2 13, 960 109, 133 ,576 pmolb 39 1 617 2 43 507 35 711 220 31 8 172 20 106 209 52 1 63 335 126 93 4077 % Compositionc 9.6 15.1 0.0 12.4 ... (6 N), and a crystal of phenol into the bottom of the hydrolysis vessel Assemble the vessel tightly Connect the vessel to a two-way argon and vacuum line Evacuate the vessel to torr, and flush...

Ngày tải lên: 08/10/2012, 10:25

... mucin (and mucin-like) glycopeptides and trifluoromethane sulfonic acid (TFMSA)-treated mucin tandem repeats to be sequenced through the heavily glycosylated regions and the macroheterogeneity to ... Lowary, T L., and Hart, G W (1996) Selective detection and site-analysis of O-GlcNAc-modified glycopeptides by β-elimination and tandem electrospray mass spectrometry Anal Biochem 234 , 38 –49 Edman ... Hoog, J.-O., and Gustavsson, A.-M., eds.), Birkhauser Verlag, Switzerland, pp 47–54 Gooley, A A and Williams, K L (1997) How to find, identify and quantitate the sugars on proteins Nature 35 8, 557–559...

Ngày tải lên: 08/10/2012, 10:25

... of the epitope and may also provide the forces between epitope and paratope required to maintain binding Conversely, it is possible to identify residues that are present in the epitope but can ... and affinity chromatography is more robust than other conventional chromatographic techniques in terms of column packing and operation (see Subheading 3. 4., Notes 13 15, and Fig 3) 1.4 General Comments ... corresponding to the MUC1 VNTR sequence synthesized according to the Pepscan approach to epitope mapping Antibodies are allowed to react with each peptide, and those containing the epitope or minimum...

Ngày tải lên: 08/10/2012, 10:25

... Etobicoke, ON, Canada) and with 100 U/mL of penicillin and 100 µL of streptomycin (Gibco-BRL) Hemocytometer Lipofectamine (Gibco-BRL) Cell culture incubator to maintain an atmosphere of 37 ˚C and ... Note 4) 3. 6 The Role of Glycosylation 3. 6.1 Tunicamycin Repeat Subheadings 3. 1. 3. 5 with the addition of 0.5 µg/mL of tunicamycin to the DMEM used in Subheading 3. 3 steps and Harvest and analyze ... mucins Glycoconj J 13, 81–90 22 Higuchi, R (1990) Recombinant PCR, in PCR Protocols (Innis, M., Gelfand, D H., and Sninsky, J J., eds.), Academic Press, Toronto, pp 177–1 83 23 Sprague, J., Condra,...

Ngày tải lên: 08/10/2012, 10:25

... HPLC Separation of Monosaccharides Reference (33 ,34 ) (35 ) (36 ) (37 ) (37 ,38 ) (39 ) (40) (41,42) (27) ( 43) (6) (44) (45) (46) 177 178 10 11 12 Michalski and Capon reductive elimination from mucus contain ... 3 C/min and then 230 to 250°C at 5°C/min Use injector and detector temperatures of 240 and 250°C, respectively Use an injection volume of µL 3. 3 Separation of Monosaccharides by HPLC 3. 3.1 Separation ... gas-liquid chromatography Anal Biochem 1 23, 33 6 34 1 26 Henri, R J., Blakeney, A B., Harris, P J., and Stone, B A (19 83) Detection of neutral and amino sugars form glycoproteins and polysaccharides...

Ngày tải lên: 08/10/2012, 10:25

... acetonitrile over 33 c 30 % acetonitrile over another 35 d Back to 20% acetonitrile over the next Store the column in 75:25 (v/v) acetonitrile:water 3. 7 HPAEC of Released Oligosaccharides 3. 7.1 ... water, lml 30 % acetic acid, and mL of acetonitrile Load the sample onto disk and leave for 15 (see Note 15) Wash the tube with 100 µL acetonitrile and add to disk Wash disk with mL of acetonitrile ... retention times to each other and to their alditols On HPAEC ( 13) , the reducing oligosaccharides will be retained significantly longer than the alditols, to give improved chromatography Sulfated...

Ngày tải lên: 08/10/2012, 10:25

... H -3 H-4 H-5 H-6' 4 .39 3 4.0 63 3.506 4.1 93 3.628 GlcNAc(β1 3) GalNAc-ol Gal(β1 3) [GlcNAc(β1–6)]GalNAc-ol 4.286 3. 995 3. 546 4.141 ND NeuAc(α2–6)GalNAc-ol 4 .39 1 4.069 3. 468 4.277 ND 4.246 3. 846 3. 411 ... 4.020 3. 532 Gal(β1 3) [NeuAc(α2–6)]GalNAc-ol H-2 H -3 H-4 H-5 H-6' GlcNAc(β1 3) [NeuAc(α2–6)]GalNAc-ol GlcNAc(β1–6)GalNAc-ol 4 .37 8 4.055 3. 534 4.244 3. 486 4.260 3. 984 ND 4.185 3. 490 4.242 3. 841 3. 379 ... Oligosaccharides 1 93 Fig Strategy of elucidation of oligosaccharide primary structure atoms; Gal H -3 and H-4 atoms; Fuc H-5 and H-6 atoms; NeuAc H-3ax and H-3eq atoms; and CH3 of the acetamido...

Ngày tải lên: 08/10/2012, 10:25

... Mucins 2 13 Fig Elution positions of acetate (CH3COO–), chloride (Cl–), sulfate (HSO4–), and phosphate (H2PO3–) ions (see also Note 11) Chromatogram displays 10 nmol each of phosphate and sulfate, and ... aliquoted and stored at –20°C, and used as required 214 Harrison and Packer Fig Quantitation of sulfation of glycopeptides A and B from human MUC2 The chromatogram shown corresponds to half of ... approx 250 mL/h of chromatography Note that carbohydrates are hygroscopic and must be thoroughly dried before weighing and making up as standards Standards of mM each of 2-, 3- , and 6-sulfo-Nacetylglucosamine...

Ngày tải lên: 08/10/2012, 10:25

... tubes (3 5 mL) with round bottom (Sarstedt, Nümbrecht, Germany) Water bath at 37 °C Methods 3. 1 Preparation of Collagen Coated Culture Flasks and Swiss 3T3 Feeder Cells (see Notes 1 3) To prepare ... CO2 in air incubator ( 13) In some situations, 3T3-conditioned medium (Subheading 2., item 5d) can be used instead of adding mouse 3T3 cells directly to cultures (see Note 5) 3. 3 Long-Term Culture ... dispase just to cover the cells, and incubate for approx 30 at 37 °C Remove the cells as a sheet, and pipet with a Pasteur pipet to remove them from the flask and to break up the sheets into smaller...

Ngày tải lên: 08/10/2012, 10:25