physician s direct obligations to the patient

... adapted thermodynamics to their needs Consequently, the successive revisions of this text incorporated examples and exercises representative of these fields In general, the spirit and format of the ... all pressures below the transition pressure and that diamond is the stable allotrope at all pressures above the transition pressure But diamond is not converted to graphite at low pressures for ... shall speak of the properties of a system, which is any region of the universe, large or small, that is being considered in our analysis Regions outside the boundaries of the system constitute the...

Ngày tải lên: 02/04/2014, 16:43

... 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 methods ... is highly unusual relative to the other eight values The problem is that both the sample mean and the sample standard deviation are 24 BASIC STATISTICS sensitive to outliers That is, the classic ... difference is always zero, so this approach is unsatisfactory Another possibility is to use the average of the absolute deviation scores: n ¯ |Xi − X | This is reasonable, but it makes certain theoretical...

Ngày tải lên: 11/06/2014, 00:51

... expensive Among them, the hot embossing process provides several advantages such as relatively low cost for the embossing tools, the simplicity of the process, the high replication accuracy for small ... 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 is crucial ... the glass transition temperature uner the isothermal conditions in the conventional hot embossing process However, the conventional hot embossing process has some inherent drawbacks such as the...

Ngày tải lên: 14/11/2014, 13:26

... Centrala Försökdjursnämnden, Swedish Match AB, the Swedish Cancer Fund, Greta och Johan Kocks Stiftelse, Stiftelsen Riksförbundet Cystisk Fibros, Vårdals Stiftelse, and the Smokeless Tobacco Research ... laboratory, CsCl rather than CsBr or CsSO4, is used as the density gradient– forming salt since gradients are run in the presence of guanidinium chloride and the use of CsBr or CsSO4 in the presence ... When mucins are isolated from tissue samples, it may be an advantage to “physically” separate histologically defined areas of the tissue such as the surface and the submucosa of an epithelium For...

Ngày tải lên: 08/10/2012, 10:25

... enzymes such as trypsin and pepsin In all cases, the success or failure of these techniques must be determined empirically Subheading 3.4 and 3.5 discuss a by-no-means exhaustive selection of these ... linkage to type backbone structures (18) Despite the previously discussed examples, lectins are not necessarily as specific in their binding affinities as is suggested in commercial data sheets or the ... other steps in the protocol identical to the test sample For immunohistochemistry, an even better approach is substitution of another antibody of the same donor species (preferably the same isotype...

Ngày tải lên: 08/10/2012, 10:25

... and comparison across blots, a standard in the same buffer as samples should be titrated for use as a standard curve, and samples should be included on all blots to determine interassay variation ... 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 more ... cancer-associated serum antigen (CASA) assay detects very high levels 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...

Ngày tải lên: 08/10/2012, 10:25

... it is less satisfactory for these tissues since the mucous gel is not so firm and the mucosa is more elastic than in the stomach The unfixed section method has been applied to human biopsy specimens ... specimens from the gastric mucosa However, it is very difficult to obtain satisfactory sections from biopsies, and hence, consistent results for mucous thickness It is necessary to obtain resected ... mucosa to use this method for human studies The distribution of mucous thickness readings on histologically stained and fixed sections is usually nonparametric Therefore, thickness values obtained...

Ngày tải lên: 08/10/2012, 10:25

... these six detailed assays are used to quantify a particular aspect of the biosynthesis of mucin These assays are essential in the extensive protocol for quantitation of the biosynthesis and secretion ... of these assays All measures of the expression of mucin are relative to parameters within the cell sample or its homogenate But, note that the use of tissue or cell samples of similar sizes makes ... proteins Therefore, it is essential to compare the [3 5S] amino acid–labeled protein bands on SDS-PAGE from all samples to identify gross alterations in protein synthesis during the experiment The specific...

Ngày tải lên: 08/10/2012, 10:25

... sequence of 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 ... data, an assessment of peptide homogeneity is advisable We analyze aliquots of the peaks by using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS), and ... charge distribution are essentially the same (8a) Analysis of Western blots with MUC-specific antisera suggest that the major mucin in this sample is the MUC5B gene product (8a) Thus, the Mono-Q...

Ngày tải lên: 08/10/2012, 10:25

... 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 is to provide ... 40Ti rotor, we typically centrifuge for 2.5 h to disperse intact mucins across the gradient and 6–8 h for the reduced subunits This is not usually performed unless there are doubts about the stability ... purified mucous extracts If we assume that the physical size and shape of the molecule are unchanged by the supporting medium and that the rotor speed is constant during the experiment, then the change...

Ngày tải lên: 08/10/2012, 10:25

... hydrolysis, the vessel should be opened as soon as it is removed from the oven Acid condensation inside the vessel is an indication of a complete hydrolysis Handling the vessel requires heat-resistant ... 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 signs that the vessel is sealed ... 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 vessel tightly...

Ngày tải lên: 08/10/2012, 10:25

... cycle is observed in the 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 ... it is possible to identify amino acids substituted with a monosaccharide using standard programs, the glycosylated amino acids are not recovered quantitatively Modifications of the transfer solvents ... heavily glycosylated peptides substituted with large oligosaccharides it may be necessary to treat with TFMSA (13) or glycosidases before sequencing There are two principle reasons for desialylating...

Ngày tải lên: 08/10/2012, 10:25

... deprotection The synthesis schedule provides details of the amounts of amino acids and activators required It is advisable to weigh out all these reagents before beginning a synthesis since this is the ... methods of simultaneous peptide synthesis, this technique allows the concurrent synthesis of hundreds to thousands of peptides so that libraries can be produced and simultaneously used as targets ... Immunoassays Both enzyme-linked immunosorbant assays (ELISAs) and radioimmunoassays have been used in various formats to test antibody binding to synthetic peptides The indirect ELISA has the advantages...

Ngày tải lên: 08/10/2012, 10:25

... Fig Schematic showing the synthesis of construct pRMC and its expression in COS cells The protocol is described in Subheadings 3.1.–3.3 Oligonucleotide primers are designed that facilitate the synthesis ... by SDS-PAGE as described in Subheading 3.5., steps 2–10 Wash the COS cells in PBS, scrape the cells into 500 µL of PBS and microfuge the suspension at 4˚C to form a pellet Resuspend the pellet ... The transfection system uses the expression vector pSVL, which is ideally suited for transfection into COS cells owing to the production of T-antigen, which increases pSVL expression (23) Dimerization...

Ngày tải lên: 08/10/2012, 10:25

... (t2 = 120 ms); E3 = –0.50 V (t3 = 300 ms) The response time is set to s Eluent Degas module to sparge and pressurize the eluents with helium (Dionex) Postcolumn with a DQP-1 single-piston pump ... hydrolysis methods generally utilized for the release of monosaccharides Consequently, the study of sialic acid must be generally considered independently of other monosaccharides The study of sialic ... Monosaccharides as Their Methylglycoside (25) Trimethylsilylated Derivatives GLC analysis allows the determination of the monosaccharide composition of glycans with amounts of total sugars not...

Ngày tải lên: 08/10/2012, 10:25

... Camberley, Surrey, UK) in series (9) Sulfated and sialylated oligosaccharide alditols can also be separated by these techniques; however, for better results the reversed-phase HPLC should be by porous ... refs 18–20 The use of electrospray ionization MS coupled with collision-induced dissociation MS is another possibility for analysis at the glycopeptide level (21) The internet provides a source ... of use The optimum volume of sample is between 50 and 100 µL Applying more or less results in poor recovery of desalted sample Likewise, sample directed down the side of the gel also results in...

Ngày tải lên: 08/10/2012, 10:25

... galactose units Since the presence of these peripheral monosaccharides affects essentially the chemical shifts of the terminal galactose, the anomeric protons can be assigned step-by-step The ... as discussed previously When the presence of these structural elements has been established, the number of possibilities decreases deeply, and a survey of the corresponding class of oligosaccharide-alditols ... increments of masses per residue are given in atomic mass units in Table 2, to this sum the mass carresponding to the number of linkage has to be substracted When sufficient material is available,...

Ngày tải lên: 08/10/2012, 10:25

... However, an excessive flow rate of sulfuric acid to the suppressor is not responsible for downward baseline drift To quantitate sulfate, it is necessary to calibrate the analysis to an external ... half of an original hydrolysis amount of 50 µg Our system for sulfate analysis uses the same chromatographic system as that for sugar analysis, in which the ion chromatography column and high-performance ... as gaseous HCl) 3.2 Analysis of Sulfate by Ion Chromatography 3.2.1 Chromatography Figure presents typical elution positions of ionic species using this method Attach the AS11 ion column to the...

Ngày tải lên: 08/10/2012, 10:25

... Methods 13 14 15 16 17 225 layer due to its size In animal tissues, smaller sections of the entire wall of the organ are used, because these tissues are more fragile and dissection is difficult The ... The presence of the muscle layer does not appear to affect the synthesis of epithelial mucins Submerging the tissue segments in the culture medium is particularly advantageous when the secretion ... refer to human colorectal cells and tissue, but similar systems and principles apply to a wide range of other tissues where mucins are produced Materials Human gastrointestinal (GI) biopsies or surgical...

Ngày tải lên: 08/10/2012, 10:25