legislation for the education of children with disabilities
chemical thermodynamics - basic concepts and methods
... constituted of the same material, and the cold object has twice the mass of the hot one, then we observe that the increase in temperature of the former is only half the decrease in temperature of the ... transformation, in the form of either heat or work, are of great interest, whether the transformation is the combustion of a fuel, the fission of a uranium nucleus, or the transport of a metabolite ... principle, the notion of the flow of a substance from the hot to the cold object appears almost intuitively, together with the concept that the total quantity of the caloric can be represented by the...
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basic statistics understanding conventional methods and modern insights jul 2009
... between the groups? The answers to these problems are nontrivial The purpose of this book is to introduce the basic tools for answering these questions The mathematical foundations of the statistical ... control for factors that are related to weight gain (the age of the rats compared) and then manipulate the single factor that is of interest, namely the amount of ozone in the air Here the goal ... called the lower and upper quartiles (The median is sometimes called the middle quartile.) Roughly, the lower quartile is the median of the smaller half of the data And the upper quartile is the...
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development of novel micro-embossing methods and microfluidic designs for biomedical applications
... was used Since the analysis of the process requires the study of various aspects relating to the characterization of the microstructures before and after the embossing, they started with flat film ... simple flow of polymer from the borders and the formation of the mounds of polymer within the cavity, which were resulted from 21 capillary action and the compression causing buckling of the polymer ... demolding, the reasons for distortion or damage of the molded part were discussed such as the adhesion at the surface, surface roughness of the mold, and the negative slopes of the cavity sidewalls The...
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Glycoprotein Methods and Protocols - P1
... and the volume of the CsCl and CHAPS must therefore be compensated for by the addition of a small volume of M guanidinium chloride The final weight of the sample is calculated from the volume of ... weight of the sample is calculated from the volume of the tube and the final density of the solution Add sodium phosphate buffer to give the final weight and stir the sample gently Measure the density ... defined areas of the tissue such as the surface and the submucosa of an epithelium For example, material from the surface epithelium may be enriched by gently scraping the surface mucosa, thereby allowing...
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Glycoprotein Methods and Protocols - P2
... best with tissues fixed in solutions other than formalin, e.g., methacarn (methanol:chloroform:acetone, 60:30:10) (44) It is beyond the scope of the current work to discuss the full range of histochemical ... U (1982) Synthesis of type human blood-group antigenic determinants: the H, X, and Y haptens and variations of the H type determinant as probes for the combining site of the lectin of Ulex europaeus ... in the case of lectin histochemistry, MAb reactivity may be modified by the removal of sialic acid (20) and neutral sugars (26) The main advantage of MAbs is in their application to the study of...
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Glycoprotein Methods and Protocols - P3
... of a glycoform of MUC1 present in the serum of pregnant women that is not detected by the CA15.3 assay (16) Similarly, these two assays show differing specificities for different glycoforms of ... Quantitation of Mucins 47 ter (2) discusses these assays and they are mentioned here only for completeness A number of general carbohydrate assays have been used for the detection of mucins, and two of the ... will differ with the application and the availability of reagents Subheading 3.2.1 outlines the procedures for preparing the membranes, and Subheading 3.2.2 describes the use of the PAS reagent...
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Glycoprotein Methods and Protocols - P4
... gel surface, the extent of which depends on the hydration of exposed mucosa and the degree of stirring of the solution above the mucosa, respectively Consequently, the dimensions of mucous thickness ... end, fill with 0.9% NaCl, and ligate the other end before snap-freezing This ensures minimum disturbance of the mucous layers and prevents contact of the adjacent luminal walls of the tissue ... and the major secretion of mucin of the gut, the adherent mucous gel is frequently ignored because of the practical difficulties in observing and quantitating it Quantitative measurement of the...
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Glycoprotein Methods and Protocols - P5
... (au) The dimensions of the various calculated values have limited significance, largely because of the use of arbitrary units for each of the measurements of radioactivity, owing to the use of the ... correlate with MUC2 protein synthesis, but that the MUC2 protein synthesis correlated highly with the total amount of MUC2 present in the tissue (6) Therefore, this study implies that MUC2 synthesis ... However, the presence of mRNA is only indirect proof of the biosynthesis of the encoded mucin This point was proven by to our knowledge, the only study that actually correlated the levels of mRNA,...
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Glycoprotein Methods and Protocols - P6
... of the effectiveness of the separation is monitored with a periodic acid-Schiff (PAS) assay and with a variety of lectins and mucin- or carbohydrate-specific antibodies (for a discussion of the ... remove the problem of peptides arising from the enzyme The amount of enzyme added was calculated assuming that protein constitutes 20% of the total mass of the mucin subunit and that approx 50% of ... example of the data obtained, using this methodology with a salivary mucin reduced subunit preparation, is shown in Fig The range of charge density of the molecules (Fig 2a) is typical of that...
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Glycoprotein Methods and Protocols - P7
... in the term r (distance from the center of rotation of the meniscus and bottom of the tube), and this information is available from the rotor data sheet Using this information together with the ... supporting medium of increasing density The sample is centrifuged and a separation of different species in the mixture is effected on the basis of their sedimentation rates The role of the preformed gradient ... µL) of the spreading agent to the surface and leave the solution for 5–15 In this time the mucins diffuse to the surface and become entrapped in the BAC monolayer Touch the carbon surface of a...
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Glycoprotein Methods and Protocols - P8
... protocol for PVDF-bound protein samples During the evacuation and argon flush of the hydrolysis vessel, the bottom of the vessel will become cold and the acid will boil under vacuum These are the ... 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 Assemble the ... 4) Place the vessel in a 105°C oven for 24 h Remove the vessel from the oven and open the vacuum tap immediately within a fume hood (see Note 4) Place the vials into a vacuum centrifuge for 10...
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Glycoprotein Methods and Protocols - P9
... bond between the carboxyl group of the sialic acid and the amine of the support Although the alternative of using diisothiocyanate-derivatized membranes is possible, we have found them of little ... Wet with 10 µL of methanol When the excess methanol has evaporated, but before the membrane has dried, apply the sample in 10-µL aliquots (see Note 12) When all of the sample is applied dry the ... methanol rather than ethyl acetate for the extraction of the cleaved amino acid from the cartridge (Fig 1, vessel 1; see Note 17) to the converter (Fig 1, vessel 2; see Note 18) Notes The arylamine-coated...
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Glycoprotein Methods and Protocols - P10
... MUC1-related antigens for the interrogation of antibody specificity; however, the methodology for their preparation is beyond the scope of this chapter By far the most widely used method for modifying ... specificity of an antibody-peptide interaction The Pepscan approach has been the most widely used and involves the synthesis of a set of overlapping peptides that span the length of the antigenic ... air-dry for c DMF to half the pin height for The next cycle of peptide synthesis can begin immediately with Fmoc deprotection When the required peptides have been synthesized, deprotect and wash the...
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Glycoprotein Methods and Protocols - P11
... the “linker” sequence and primes the 5' end of the target domain of rat Muc2: a 5'-GGCTTGGAATCTAACTGTGAAGTTGCTGC-3' Dimerization of Domains of Mucin 145 Fig Schematic showing the synthesis of ... incubate the reaction at 72˚C to allow extension of the linked templates Add µg each of primers and 6, and continue the PCR with 30 cycles at 94˚C for 30 s, 50˚C for and 72˚C for min, ending with ... H2O for mock-transformation control), stir gently with a pipet tip, and incubate the cells on ice for 30 Heat shock the reactions in a 37˚C water bath for 20 s, and then incubate them on ice for...
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Glycoprotein Methods and Protocols - P12
... these conditions, the hydroxyl groups of carbohydrates are converted to oxyanions with pKa values in the range of 12–14 The anomeric hydroxyl group of the reducing sugar is more acidic than the ... Wash the column successively with mL of 10 mM , mL of M and mL of M formic acid h Pool the fractions and lyophilize 3.1.2 Enzymatic Release of Sialic Acid Resuspend 1–10 mg of mucin in mL of 100 ... increase the sensitivity of detection As described under HPLC, the most common tagging methods are based on the reductive amination procedure, wherein the reducing end of the sugar reacts with the...
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Glycoprotein Methods and Protocols - P13
... Re-N-acetylate the glycans on the column by adding 1.4 mL of reagent C for 30 at room temperature Wash the column with × mL of reagent B followed by mL of methanol (see Note 6) Elute the oligosaccharides with ... HPLC Column Wash the column with water for 30 at a flow rate of 0.4 mL/min Wash the column with acetonitrile for 30 at a flow rate of 0.4 mL/min 186 Hounsell Wash the column with 0.5 M ammonium ... with × mL of reagent D Complete the re-N-acetylation with 0.1 mL of acetic anhydride for 30 at room temperature Wash a Sep-Pak C18 cartridge with mL of methanol and mL of H2O 10 Transfer the sample...
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Glycoprotein Methods and Protocols - P14
... resonances of the GalNAc-ol unit Moreover, the quadruplet of the H-6' signal of GalNAcol is upfield shifted out of the bulk at δ ~ 3.50 ppm in the case of an O-6 substitution with sialic acid The presence ... atoms; and CH3 of the acetamido groups The first step of spectrum analysis consists of the identification of the core region (Table 1), based on the characteristic chemical shifts of the H-2 and ... members of a series of closely related sequences, as it is happily the case for most of O-glycans When the material is available in the range of 0.1–5 µmol, the de novo structural elucidation of the...
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Glycoprotein Methods and Protocols - P15
... the AS11 ion column to the high-performance liquid chromatograph Attach the ion suppressor downstream of the column, and the conductivity cell downstream of the ion suppressor, according to the ... detection of sulfate by ion chromatography as described here is about nmol Since mucins are often highly sulfated (i.e., the number of moles of sulfate >> the number of moles of protein), the amount of ... baseline over the course of the separation method Note that the flow rate of sulfuric acid to the ion suppressor does not need to be precise; in our laboratory the flow of sulfuric acid to the suppressor...
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Glycoprotein Methods and Protocols - P16
... sequence of changes in the epithelium allows precise studies of how the changes in cells influence the mucins that are produced Several of the end products of malignant transformation of colonic epithelium ... guish them from the cells in vivo On the other hand, the cell lines may consist of only one cell type, which can be an advantage Study of the colon has the advantage that the sequence of changes ... which the production of mucin is being assessed, or removing the 3T3 feeders from the flask once the epithelium has grown In addition, the 3T3 cells should be examined for the production of mucin...
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Glycoprotein Methods and Protocols - P17
... migration of the sample into the gels (8) 17 Markers for agarose gel electrophoresis reflect the separation of molecules on the basis of their size and charge This is in contrast to the normal ... glycoproteins The metabolic rate of the cells should be considered for optimum labeling with the radioactive precursor during continuous labeling cultures The type of isotope will govern the amount ... labeled with the latter compounds The origin(s) of the differences in labeling efficiency among these organs is not known It is essential that incubation commences within 10 of removal from the body...
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