introduction to developmental psychology

... We have also made a definite effort to keep this volume to a manageable size Too often, a textbook that attempts to be exhaustive in its coverage merely serves to overwhelm the student On the other ... principles We have taken the opportunity, therefore, to revise our approach to some topics and to add problems that reflect new applications We have continued to take advantage of the resources available ... is INTRODUCTION “thermodynamics” insofar as applications to chemistry are concerned More commonly, one affixes the adjective “chemical” to thermodynamics to indicate the change in emphasis and to...

Ngày tải lên: 02/04/2014, 16:43

... factor that is of interest, namely the amount of ozone in the air Here the goal is not so much to explain how best to design an experiment but rather to provide a description of methods used to ... height 1/n A histogram is one way of trying to deal with this problem It is similar to plots of relatively frequencies, the main difference being that values are binned together to get a more useful ... goal here is to illustrate what might go wrong and provide information about where to look for better techniques First we consider a situation where the histogram tends to perform tolerably well...

Ngày tải lên: 11/06/2014, 00:51

... range from embossing to deembossing temperature) should be 25 to 40oC in order to minimize the thermally induced stresses The embossing pressure is around 0.5 to kN per cm2 Automated de-embossing ... Thanks also go to my collaborators, Dr Hank Wu, Dr Chu-hua Chen, and all other friends in Ritek, Taiwan, on the CD-ELISA project To all the fellow graduate students in our lab, especially to those ... heating method to achieve the reduced cycle time comparable to that micro-injection molding by rapid heating to soften or melt the polymer on the surface while pressing it against a mold to form the...

Ngày tải lên: 14/11/2014, 13:26

... the running 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 ... 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 analyses 3.5 Ion-Exchange Chromatography ... 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 subject to the...

Ngày tải lên: 08/10/2012, 10:25

... anatomy in sufficient detail or fail to heed variation that may be owing to differences between anatomical regions or genetic factors Relationship of the distribution of mucin should be linked to ... sections to restore antigenicity, a number of “antigen retrieval” or “antigen unmasking” techniques relying on heat to “unfix” tissues have been rapidly incorporated into the routine histochemical ... sugar and incubate for 30 to h Proceed with histochemistry protocol as usual 3.2.3 Enzymatic Deglycosylation to Confirm Lectin Specificity See Subheadings 3.6.1.–3.6.4 Histological Methods for Detection...

Ngày tải lên: 08/10/2012, 10:25

... ed.), 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, ... become possible to target probes at less glycosylated portions of the molecules; however, a drawback of these probes is that their epitopes tend to be cryptic and need reduction to be exposed In ... using antibody capture 52 McGuckin and Thornton In this protocol, purified mucin-reactive antibodies or lectins are coated onto microtiter plates and used to capture mucins in biological samples...

Ngày tải lên: 08/10/2012, 10:25

... (down to µm) for histological processing the better, because this provides clearer details of cell structure It may be necessary, in some cases, to use thicker sections (up to 20 µm) in order to ... humans is given in Newton et al (30) This histological and fixation method can be adapted to staining the adherent mucous gel with biotinylated antibodies and lectins coupled to avidin-horseradish ... experimentation to ensure full maintenance of in vivo functions during the experimental period, can be found in refs 20–22 Various groups have adapted histological procedures to observe adherent...

Ngày tải lên: 08/10/2012, 10:25

... strongly advised to run a sample of the specifically immunoprecipitated mucin precursor of interest on the same gel, to be able to identify unequivocally the mucin precursor prior to quantitation ... tissue to produce mucins (1,4–11) More important, a distinction can be made with the preexisting, stored mucin in the mucin-producing cells Metabolic labeling during short periods (up to 60 min) ... short periods (up to 60 min) will not add significant mucin to the vast reservoir of stored mucins Therefore, researchers will be able to distinguish within one experiment the movements of two...

Ngày tải lên: 08/10/2012, 10:25

... reduction (Mr = 2–3 × 106) are amenable to conventional biochemical separation methods We describe a separation protocol based on anion exchange chromatography to fractionate the different species ... in respiratory mucin subunit preparations In respiratory samples there are typically two gel-forming mucins, namely MUC5AC and MUC5B The MUC5B mucin appears by Mono-Q chromatography to be in two ... Thornton et al 3.3.1 Anion-Exchange Chromatography The flow rate for anion-exchange is 0.5 mL/min throughout the chromatography, and typically 0.5-mL fractions are collected Apply the sample (up to...

Ngày tải lên: 08/10/2012, 10:25

... 1.3 mL/min/tube (see Note 6) Weigh tubes to check that they contain equal volumes Carefully apply sample (up to 500 µL) with a Hamilton syringe to the top of the gradient (see Note 7) Centrifuge ... obtained from a respiratory mucin preparation 90 Sheehan and Thornton Fig Rate-zonal centrifugation of respiratory mucous extract A respiratory mucous extract in M GuHCl was applied to a 6–8 M GuHCl ... solvent is 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...

Ngày tải lên: 08/10/2012, 10:25

... adjusted to pH 8.5 with NaOH Buffer may be kept up to mo at 4°C Fmoc reagent: mg/mL fluorenylmethyl chloroformate (Sigma, cat no F0378) in acetonitrile (HPLC grade) Reagent may be stored up to wk ... solution of a sample of mucin into an autosampler glass vial and dry under vacuum (see Note 3) Place the vials, 400 µL of HCl (6 N), and a crystal of phenol into the bottom of the hydrolysis vessel ... be dot-spotted onto polyvinylidene difluoride (PVDF) membrane, or gel separated by electrophoresis and electroblotted onto PVDF Note that mucins are often difficult to bind to PVDF Wilkins et...

Ngày tải lên: 08/10/2012, 10:25

... tandem repeats to be sequenced through the heavily glycosylated regions and the macroheterogeneity to be assigned and quantitated (9–11) Simple modifications to most modern sequenators facilitate ... sequencer An alternative approach is to use solid-phase sequencing to extract glycosylated amino acids (5) Peptides and proteins are covalently attached to solid supports (6,7), allowing the ... modified Beckman LF3600 sequenator (12) These methods were developed for two reasons: (1) to provide a distinct elution window for the PTH-glycosylated amino acids, and (2) to be of low enough salt...

Ngày tải lên: 08/10/2012, 10:25

... 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 ... 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 ... ultraviolet monitor has returned to baseline Optional: Wash the column with 0.5 M NaCl (1 mL/min) to remove material that has bound to the column nonspecifically 138 Murray et al To achieve desorption...

Ngày tải lên: 08/10/2012, 10:25

... (FBS) (CanSera, 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 ... DNA encoding the domain of interest is ligated to a known epitope sequence (for detection by a suitable antibody), and to a signal peptide sequence (to ensure secretion), by recombinant polymerase ... bromophenol blue, plus or minus 1,4-Dithiothreitol (DTT) to give a final concentration of 10 mM Filter concentrators such as Centricon with a molecular weight cutoff of 30 kDa Phenylmethylsulfonylfluoride...

Ngày tải lên: 08/10/2012, 10:25

... separated without introduction of base in the eluent Because the detector works only at high pH, postcolumn addition of NaOH to the effluent before the entry into the PAD detector is required (7) ... of success) amounts of glycoproteins or oligosaccharides corresponding to 10 µg of total sugar to which µg of mesoinositol is added as an internal standard Add 0.5 mL of methanol–HCl reagent ... methylglycosides to GLC 3.2.4.2 GAS CHROMATOGRAPHY CONDITIONS A typical GLC chromatogram of TMS derivatives is given in Fig Neutral monosaccharides generally provide several peaks corresponding to pyrano,...

Ngày tải lên: 08/10/2012, 10:25

... mL/min with water/ acetonitrile containing 0.2% TFA: a 20% acetonitrile for b 40% acetonitrile over 33 c 30% acetonitrile over another 35 d Back to 20% acetonitrile over the next Store the column in ... 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 ... (see Note 8): a 75% acetonitrile for b 62.5% acetonitrile over the next 25 c 60% over the next 30 d Back to 75% at 60 for the total run 3.6.2 Porous Graphitized Carbon Chromatography Wash the column...

Ngày tải lên: 08/10/2012, 10:25

... the diagonal Two-dimensional total correlation spectroscopy (2D 1HTOCSY) can be used to characterize all the proton resonances A transfer of magnetization from H-1 to H-6 is observed in the case ... 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 groups The first step of spectrum ... NeuAc attached to the terminal galactose units Since the presence of these peripheral monosaccharides affects essentially the chemical shifts of the terminal galactose, the anomeric protons can be...

Ngày tải lên: 08/10/2012, 10:25

... centrifuging to spin condensate to the bottom of the tube, transfer the hydrolyzed samples to a vacuum centrifuge and dry completely When dry, add 50 µL of water to each sample and dry again, in order to ... Ion Chromatography 3.2.1 Chromatography Figure presents typical elution positions of ionic species using this method Attach the AS11 ion column to the high-performance liquid chromatograph Attach ... (70% water, 30% 0.1 M NaOH) b 10 to 11 min: isocratic gradient (curve = 5) at 30 mM NaOH c 11 to 15 min: curved gradient (curve = 1) from 30 mM NaOH to mM NaOH d 16 to 25 min: re-equilibration in...

Ngày tải lên: 08/10/2012, 10:25

... streptomycin, and put onto 24-h postconfluent 3T3 cell layers for 24 h After conditioning, the medium is filtered through a 0.2-mm filter (Nalgene, Milton Keynes, UK) and further supplemented to ... 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 ... Proliferating cells in the bottom of the colonic crypts migrate to the upper half of the crypts, where they differentiate These differentiated cells migrate to the top of the crypt, and there is...

Ngày tải lên: 08/10/2012, 10:25