bluuebunny/crossref_metadata_2025_split huggingface dataset:
Structural determination of oligosaccharides from recombinant iduronidase released with peptide N‐glycanase F using fluorophore‐assisted carbohydrate electrophoresis
Authors: Hague Chuck , Masada R. Irene , Starr Christopher 
Published: May 2005

AbstractThe lysosomal storage disorder mucopolysaccharidoses I (MPS I) is caused by a deficiency in the production of α‐L‐iduronidase. Recently, a recombinant α‐L‐iduronidase has been produced in Chinese hamster ovary (CHO) cells. It is thought that for α‐L‐iduronidase to be correctly targeted to the lysosomal vesicle a particular oligosaccharide make‐up must be present, and characterization of the carbohydrates is critical. Oligosaccharides from α‐L‐iduronidase were analyzed using fluorophore‐assisted carbohydrate electrophoresis (FACE®). The FACE® system uses polyacrylamide gel electrophoresis to separate, quantify, and determine the sequence of oligosaccharides released from glycoproteins. Asparagine‐linked oligosaccharides were released from α‐L‐iduronidase using the enzyme peptide N‐glycanase F (PNGase F). Released oligosaccharides were labeled with a fluorophore at the reducing termini by reductive amination. A total of nine bands were sequenced from the released pool of oligosaccharides. The pool of fluorescently labeled oligosaccharides was then electrophoresed in preparative gels and each band individually excised and extracted. Isolated bands were treated with a series of exoenzymes to determine the sequence of monosaccharides that make up a particular oligosaccharide. A total of eighteen different oligosaccharides were identified from the original pool of oligosaccharides. A majority of the oligosaccharides, over 73%, were found to be of the sialylated complex type. Four of the oligosaccharides were phosphorylated, making up approximately 11% of the carbohydrate pool, and the remaining 15% were of the oligomannose type.
Electrophoretic behavior and size distribution of the acidic polysaccharides produced by the bacteria Bradyrhizobium (Chamaecytisus) strain BGA‐1 and Bradyrhizobium japonicum USDA 110
Authors: Díaz‐Marrero Ana R. , Santamaria Mónica , Poveda Ana , Jiménez‐Barbero Jesus , Corzo Javier 
Published: May 2005

AbstractThe electrophoretic behavior in polyacrylamide gels of the acidic polysaccharides produced by the soil bacteria Bradyrhizobium (Chamaecytisus) strain BGA1 and Bradyrhizobium japonicum USDA110 has been studied. Both polysaccharides were polydisperse, producing a ladder‐like pattern after fixation with Alcian Blue and silver staining of the gel. The polysaccharide molecules were separated according to their size, and they behaved as a collection of flexible random coils of different size and similar charge/mass ratio. The electrophoretic behavior was not affected by the presence of acetyl groups in the polysaccharide. The range of molecular weights of the exopolysaccharide produced by B. japonicum USDA110 was wider and with larger molecules than that of the polysaccharide produced by strain BGA1. The resolution was dependent on the electrophoresis buffer; the best results were achieved with Tris‐borate; in Tris‐glycine buffer, the resolution was worse, and it was not improved by the addition of sodium dodecyl sulfate (SDS).
Profiling glycoprotein N‐linked oligosaccharide by capillary electrophoresis
Authors: Chen Fu‐Tai A. , Evangelista Ramon A. 
Published: May 2005

AbstractA method for analysis of N‐linked oligosaccharides derived from glycoproteins including sialic acid‐containing species is presented. It is based on the combination of specific chemical and enzymatic conversions coupled with capillary electrophoretic (CE) separation and laser‐induced fluorescence (LIF) detection. Glycoproteins were heat‐denatured in the presence of a reducing agent and the N‐linked oligosaccharides were released by peptide N‐glycosidase (PNGase F; EC3.5.1.52)‐catalyzed hydrolysis. The released N‐linked oligosaccharides were derivatized with 8‐aminopyrene‐1,3,6‐trisulfonate (APTS) under mild reductive amination conditions in which desialylation and loss of fucose residues are minimized. A model N‐linked oligosaccharide, desialylated, galactosylated biantennary, core‐substituted with fucose (A2F) was tested for APTS‐based derivatization chemistry with excellent recovery of the adduct without losing fucose and neuraminic acid residues. The profiles of heavily sialylated N‐linked oligosaccharides derived from fetuin, recombinant human erythropoietin and kallikrein are reported and the data show that the present method produces a high resoluton of the N‐linked oligosaccharide profile for fingerprinting glycans derived from glycoproteins.
Enhancement of sample loadings for the analysis of oligosaccharides isolated from Pseudomonas aeruginosa using transient isotachophoresis and capillary zone electrophoresis—electrospray—mass spectrometry
Authors: Auriola Seppo , Thibault Pierre , Sadovskaya Irina , Altman Eleonora 
Published: May 2005

AbstractThe analysis of underivatized core oligosaccharides arising from mild acid hydrolysis of lipopolysaccharides from Pseudomonas aeruginosa serotype O5 was achieved using a transient isotachophoretic preconcentration method coupled to capillary zone electrophoresis‐electrospray‐mass spectrometry (tCITP‐CZE‐ES‐MS). The combination of a tCITP preconcentration step provided a 10‐ to 50‐fold enhancement of sample loading and a corresponding improvement in sensitivity compared to the conventional zone electrophoresis format. Electrophoretic conditions, enabling the separation of these anionic analytes, were developed to determine possible sites of heterogeneity on either the core or the O‐chain structures. The tCITP‐CZE‐ES‐MS technique provided unparalleled resolution of the different core glycoforms and oligosaccharides obtained from the acid cleavage of the native endotoxins whether isolated following conventional gel permeation chromatography or obtained from direct hydrolysis of the bacterial isolates. These investigations also highlighted the highly phosphorylated nature of these complex cell membrane components, where the heptose residues of the core oligosaccharide can bear up to six phosphate groups# NRCC: 42370.
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Polymerase chain reaction—single strand conformation polymorphism or how to detect reliably and efficiently each sequence variation in many samples and many genes
Authors: Jaeckel Silke , Epplen Jörg T. , Kauth Marion , Miterski Bianca , Tschentscher Frank , Epplen Cornelia 
Published: May 2005

AbstractA simple and fast method with high reliability is necessary for the identification of mutations, polymorphisms and sequence variants (MPSV) within many genes and many samples, e.g. to clarify the genetic background of individuals with multifactorial diseases. We evaluated polymerase chain reaction—single strand conformation polymorphism (PCR‐SSCP) analysis to identify MPSV in several genes, which are thought to be involved in the pathogenesis of multifactorial autoimmune diseases like multiple sclerosis. The method is based on the property, that the electrophoretic mobility of single‐stranded nucleic acids depends not only on its size but also on its sequence. The target sequence was amplified, digested into fragments ranging from 50–200 bp, heat‐denatured and analyzed on native gels. The analysis of 55 PCR systems, including a total of 145 fragments demonstrates, that the detection rate of MPSV depends primarily on the fragment lengths. Appropriate dilutions of samples enhances the proportion of ssDNA compared to dsDNA. Changing the gel conditions, glycerol concentrations and/or the addition of urea may increase fragment resolution in some cases. In general, the detection of MPSV is neither influenced by their location within the fragment nor by the type of substitution, i.e. transitions or transversions. The standard PCR‐SSCP system described here provides high reliability and detection rates and allows the efficient analysis of many samples and many genes.
Multiplexed short tandem repeat polymorphisms of the Weber 8A set of markers using tailed primers and infrared fluorescence detection
Authors: Oetting William S. , Armstrong Catherine M. , Ronan Shawn M. , Young Terri L. , Sellers Thomas A. , King Richard A. 
Published: May 2005

AbstractShort tandem repeat polymorphism (STRP) markers have become important reagents for mapping genetic diseases. These markers are available as screening sets, which are located in all chromosomes at discrete intervals, allowing the entire genome to be analyzed. Mapping studies that include many individuals in the analysis necessitate the production of large numbers of genotypes. In an effort to increase the efficiency and lower the cost of using these STRP screening sets, we have divided the amplification primers of the Weber 8A screening set into groups that can be amplified in single polymerase chain reaction (PCR) amplification reactions, resulting in a reduction of both time and cost. Fluorescently‐labeled amplification products were produced using a three primer reaction. The forward STRP amplification primer for each marker contained a 19 bp sequence at the 5′ end. A fluorescently‐labeled primer, with a sequence identical to the 19 bp tail, was added to the amplification reaction as the sole source of fluorescent label. The STRP banding pattern is detected using an automated fluorescent DNA sequencer. Use of this multiplexed genomic screening set should greatly enhance the mapping of human disease loci.
Capillary electrophoretic analysis of synthetic short‐chain oligoribonucleotides
Authors: Cellai Luciano , Onori Alberto Mochi , Desiderio Claudia , Fanali Salvatore 
Published: May 2005

AbstractThirty synthetic oligoribonucleotides, 3 to 18 nucleotides (nt) long, were analyzed by capillary electrophoresis, under nondenaturing conditions, using a commercial kit. The migration time tm was dependent on nt length and composition, capillary length, operating temperature, and type of sieving polymer. Under fixed experimental conditions, the tm proved predictable by the equation: tm = [0.22(n‐1) + 6.14A/n + 6.86G/n + 3.61 (C+U)/n] min, for n3, where A/n, G/n, C/n, U/n is the frequency of each type of nt within the oligonucleotide (ONT). The equation accounts for the influence of charge‐to‐mass ratio on tm, but not for structural effects, if present. This approximation is acceptable for short ONTs. The possibility of detecting n+1, n‐1, n‐2 impurities, having predicted the tm, is of crucial importance in assessing the purity of synthetic ONTs dedicated to structural studies. This appears to be feasible. High resolution was shown among homologous series of ONTs of increasing length, and in some cases, even within groups of ONTs of the same length but different composition. The addition of 7 M urea to the buffer, as denaturing agent, accelerates the tm and significantly lowers the resolution for the shortest ONTs. It was also possible to monitor the state of association of mixtures of RNA and DNA sequence‐complementary strands.
Nonaqueous capillary electrophoretic separation and thermo‐optical absorbance detection of five tricyclic antidepressants and metabolism of amitriptyline by Cunninghamella elegans
Authors: Li Xing‐Fang , Liu Chung‐Sheng , Roos Pieter , Hansen Eugene B. , Cerniglia Carl E. , Dovichi Norman J. 
Published: May 2005

AbstractWe developed a technique based on nonaqueous capillary electrophoresis and laser‐based thermo‐optical absorbance detection to assay five antidepressants with similar structures and mass‐to‐charge ratios. A mixture of methanol and acetonitrile with ammonium acetate was essential to achieve baseline resolution of these compounds. We investigated the effects of ammonium acetate concentration, temperature, applied voltage, and capillary length on separation efficiency. The nonaqueous capillary electrophoresis and laser‐based thermo‐optical absorbance detection technique was used to study the metabolism of amitriptyline by Cunninghamella elegans. Sample preparation procedures were simplified for fast screening of the parent drug and its metabolites. Reproducible electropherograms were obtained from replicate cultures of C. elegans growing in the presence of amitriptyline.
Cholangiocyte‐specific rat liver proteins identified by establishment of a two‐dimensional gel protein database
Authors: Tietz Pamela , de Groen Piet C. , Leigh Anderson N. , Sims Christina , Esquer‐Blasco Ricardo , Meheus Lydie , Raymackers Jos , Dauwe Martine , Larusso Nicholas F. 
Published: May 2005

AbstractThe liver is composed of a variety of cells that form a functional unit involved in uptake, synthesis, metabolism, and secretion. Until recently, most studies examining liver function did not analyze the specific proteins expressed or functions performed by the multiple individual cell types that constitute the hepatic mass. In the last decade, novel isolation methods have been developed that allow the purification of liver cell populations highly enriched in one type of liver cell. Here, we present a detailed two‐dimensional (2‐D) protein map of rat bile duct epithelial cells (i.e., cholangiocytes) using a recently developed isolation procedure. In addition, we identify 27 major cholangiocyte proteins either by comparison to maps of known rat liver proteins (based on pI and Mr) or by tryptic digestion and microsequencing. Finally, we compare the relative abundance of individual proteins present in cholangiocytes to whole liver as well as hepatocyte‐specific proteins. Our results show that cholangiocytes express a unique array of individual proteins. The cholangiocyte 2‐D protein pattern is markedly different from that of isolated rat hepatocytes or whole rat liver, with high levels of proteins previously known to be expressed by cholangiocytes (e.g., cytokeratins, actins) as well as protein not previously demonstrated to be expressed at high levels (e.g., annexin V, selenium binding protein). We conclude that this cholangiocyte‐derived, 2‐D protein map will be a crucial resource for studies directed at our understanding of cholangiocyte physiology and pathobiology.
Potential heat: Considerations in the application of a new fire test
Authors: Robertson A. F. 
Published: May 2005

AbstractA review is presented of the nature of the potential heat test method. It is emphasized that the procedure usually yields a property‐type measurement of the material or composite when applied. As a result, care must be taken to insure that necessary supplementary tests are performed by other methods to guarantee that a substantial fraction of the possible fire hazard has been considered. Suggestions for appropriate use of the procedure are included and information available on the precision of the procedure is reviewed. It is concluded that, when properly applied, the procedure can serve well as one of the tools for characterizing possible fire hazards. Suggestions are made for possible future development of the procedure to yield information on rate of heat release.
Burning plastics: Mechanisms of smoke formation
Authors: Green R. J. S. , Hume J. , Kumar S. 
Published: May 2005

AbstractQuantitative pyrolysis gas chromatography studies have been conducted on three grades of cross‐linked polyester resin formulated for improved resistance to surface spread of flame. Pyrolysis temperatures from 723 to 1273 K were used, and these values correspond to ‘surface’ temperatures of materials exposed to heat fluxed of 15–149 kW m–2 which are typical of conditions experienced in real fires. Comparison is made with earlier macro studies on the evolution of smoke from plastics materials exposed to similar heat fluxes under an inert atmosphere. There is some correlation between the smoke obscuration data obtained in previous macro studies and the yield of volatile aromatic products evolved on pyrolysis. The presence of flame retardants increases the formation of carbon in the pyrolysis residue and decreases the yield of volatile aromatic products isolated. The results are considered in the context of mechanisms of smoke formation.
Furniture and furnishings in the home—some fire statistics
Authors: Chandler S. E. , Baldwin R. 
Published: May 2005

AbstractA statistical study of fires in the United Kingdom involving the ignition of furniture and furnishings is presented. This paper examines the data for one year (1970). The analysis shows that in fires starting in furniture and furnishings the chance of a fatality is over twice that in other domestic fires. The majority of furniture fires involve upholstery or bedding and over 90% were started by smokers' materials, electric appliances, space heating or as the result of the activities of children or suspected arsonists. Eighty‐five percent of the fatalities were found in the room of origin of the fire. Eighty per cent were overcome by smoke or toxic gases. Sixty percent of the fatalities were either under 5 or over 65 years of age. Monetary values are assigned for damage, casualties and deaths in fire. These costs can be used to assess the value of fire precautions. With the values taken, the total losses in furniture fires in the home amounted to £19 million in 1970. Life loss accounted for the major part of this sum. The expected annual loss per dwelling as a result of the ignition of furniture is thus only about £1, and is only £3 for all dwelling fires. This low figure suggests an approach of either selective spending on those most at risk (the elderly and handicapped) or by government activity through publicity and education.
Quantification of threat from a rapidly growing fire in terms of relative material properties
Authors: Friedman Raymond 
Published: May 2005

AbstractEquations have been developed which give the time available for escape or rescue, i.e., the time interval between detection and blockage of the escape route by smoke, heat or toxic gases. Alternative assumptions are explored concerning exponential vs power law fire growth and an extended fire plume vs uniform filing of the building. The equations are developed in such a form that the threat variable by which the fire is detected is not necessarily the same threat variable which first blocks the escape route. A number of interesting results have been obtained, and numerical values of key parameters measured in various test fires at Factory Mutual Research are tabulated. It is shown that for many polymeric fuels smoke will block the escape route well before temperature or toxicity becomes excessive. In such cases, if the fire, assumed to be growing exponentially, is detected by its smoke, the detector being located in the escape route, then the escape time, surprisingly, is independent of the smokiness of the material as well as the size and shape of the building. It is determined only by the growth rate constant (doubling time) of the fire and the sensitivity of the detector.
Post‐flashover compartment fires: Basis of a theoretical model
Authors: Babrauskas Vytenis , Williamson Robert Brady 
Published: May 2005

AbstractA theoretical model of compartment fires in the post‐flashover stage is presented. The model incorporates the stirred reactor assumption and window flow approximation of Kawagoe, but treats fuel burning rates and burning regimes in a theoretical manner. The resulting formulation preserves the important features of the fire behavior but is simple enough to permit use for building fire safety design purposes.
Quantification of smoke produced at fires. Test methods for smoke and methods of expressing smoke evolution
Authors: Rasbash D. J. , Phillips R. P. 
Published: May 2005

AbstractA limited review of the methods available for measuring smoke from burning materials has been carried out in order to define a method that could provide data for calculating smoke load. Results available in the literature have been expressed on a common basis and augmented by further experimental work carried out with a furnace similar to the National Bureau of Standards test furnace and a smoke‐containment volume (13.4m3) closer to that used by the Fire Research Station test. Encouraging agreement between the data have been found for methods which allow smoke to become diluted and to accumulate. Agreement could be improved if an allowance was made for flaming being extinguished at different times in a test, dependent on thermal stress and oxygen concentration, even when a pilot light is present. To help express the results clearly, a new unit for smokiness is proposed—the obscura (ob) defined as 1 ob=1 dB light attenuation per metre of light path. The smoke‐producing potential of different materials (Do) is then expressed in units of obscura—cubic metres per gram of volatiles produced during the fire condition (ob m3 g −1).
Heat transfer from a buoyant smoke layer beneath a ceiling to a sprinkler spray. 1—a tentative theory
Authors: Morgan Howard P. 
Published: May 2005

AbstractA theory is developed for calculating the heat transferred from a buoyant layer of fire gases and smoke, to a sprinkler spray. The theory involves calculating the heat transfer to a single water drop as it describes its trajectory, and uses experimentally derived information on the nature and structure of such sprays to calculate heat transfer to the whole spray. Because such experimental information is sparse for sprinklers, a very simple model of the ballistic properties of a sprinkler spray is adopted. Calculations using the teory suggest that the practive of installing sprinklers in the smoke reservoirs of shopping malls would, in some cases at least, reduce the effectiveness of natural venting of smoke by reducing the buoyancy of the hot smoky gases.
Thermal conditions to produce skin burns
Authors: Bull J. P. , Lawrence J. C. 
Published: May 2005

AbstractThe numbers and severities of burn casualties are described by reference to national data and a more detailed analysis of causes of burns in Birmingham. The important dimensions of any burn are area and depth, both are relevant to severity of injury and mortality. Personal factors, such as age, disability, unconsciousness, drugs and alcohol, can influence depth and area of burning. There is a relation between exposure time and thermal flux to cause burning, this is described for hot metal and radiant heat. The data can be used to predict likelihood of sustaining burns in specified situation. Such prediction is less easy with poor heat conductors; the reasons for this are discussed. The relevance of this information for advising on safety in a variety of situations is outlined.