bluuebunny/crossref_metadata_2025_split huggingface dataset:
Proteome analysis: Biological assay or data archive?
Authors: Haynes Paul A. , Gygi Steven P. , Figeys Daniel , Aebersold Ruedi 
Published: May 2005

AbstractIn this review we examine the current state of proteome analysis. There are three main issues discussed: why it is necessary to study proteomes; how proteomes can be analyzed with current technology; and how proteome analysis can be used to enhance biological research. We conclude that proteome analysis is an essential tool in the understanding of regulated biological systems. Current technology, while still mostly limited to the more abundant proteins, enables the use of proteome analysis both to establish databases of proteins present, and to perform biological assays involving measurement of multiple variables. We believe that the utility of proteome analysis in future biological research will continue to be enhanced by further improvements in analytical technology.
The Australian proteome analysis facility (APAF): Assembling large scale proteomics through integration and automation
Authors: Walsh Bradley J. , Molloy Mark P. , Williams Keith L. 
Published: May 2005

AbstractThe field of proteomics opens new possibilities for the mass screening of proteins from many different sources. While genomics is well understood to be a big science field, proteomics is just emerging as such. This paper describes the setting up of the first national proteomics facility. The facility has been funded by the Australian government and this funding has allowed the design of purpose built, integrated laboratories with state of the art equipment for large scale proteome research.
New insights into cyclosporine A nephrotoxicity by proteome analysis
Authors: Aicher Lothar , Wahl Daniel , Arce Agustin , Grenet Olivier , Steiner Sandra 
Published: May 2005

AbstractUsing two‐dimensional gel electrophoresis (2‐DE), we recently discovered an association between decreased calcium‐binding protein, calbindin‐D 28 kDa, urinary calcium wasting and intratubular corticomedullary calcifications in rat kidney. This observation prompted us to investigate kidney tissues of other species, including man. In this paper we show that in dogs and monkeys, which are generally devoid of cyclosporine A (CsA)‐mediated nephrotoxicity, renal calbindin levels were not affected by the CsA treatment whereas in CsA‐treated human kidney‐transplant recipients with renal vascular or tubular toxicity, a marked decrease in renal calbindin‐D 28 kDa protein level was found in most of the kidney biopsy sections. The present results strongly suggest that calbindin is a marker for CsA‐nephrotoxicity. The discovery of calbindin‐D 28 kDa being involved in CsA toxicity has evolved from the application of 2‐DE and has not been reported previously, proving that proteomics can provide essential information in mechanistic toxicology. Considering the current improvements in proteome methods it is expected that high throughput proteomics will become an indispensable tool in preclinical safety testing.
Charge heterogeneity of macrophage migration inhibitory factor (MIF) in human liver and breast tissue
Authors: Magi Barbara , Bini Luca , Liberatori Sabrina , Marzocchi Barbara , Raggiaschi Roberto , Arcuri Felice , Tripodi Sergio A. , Cintorino Marcella , Tosi Piero , Pallini Vitaliano 
Published: May 2005

AbstractMacrophage migration inhibitory factor (MIF) is an ubiquitous protein playing various immunological and hormonal roles. Theoretical electrophoretic coordinates calculated from protein sequence in the SWISS‐PROT database (AC P14174) are 12 kDa and pI 8.24. Using two‐dimensional (2‐D) immunoblotting, we have detected isoelectric forms at ca. 11.9 kDa, with pI values of 7.8 and 6.98 in human liver tissue, breast tissue and a cell line and in preparations of human MIF expressed in E. coli. This evidence suggests that MIF charge heterogeneity originates from a post‐translational modification not requiring euka‐ryote‐specific enzymes. We have also detected in human liver a minor immunoreactive spot at pI 6.23, which coincides with the MIF spot in the liver map in SWISS‐2DPAGE. The pI 6.23 isoform also conceivably derives from post‐translational modification, as MIF is known to be encoded in the human genome by a single copy gene.
Micellar electrokinetic chromatography: A convenient alternative to colorimetric and high performance liquid chromatographic detection to monitor protease activity
Authors: Viglio Simona , Zanaboni Giuseppe , Cetta Giuseppe , Iadarola Paolo , Luisetti Maurizio , Guglielminetti Maria 
Published: May 2005

AbstractHigh performance capillary electrophoresis (HPCE) has been exploited as an analytical method alternative to current procedures for the determination of proteolytic activity of elastases from different sources. Due to some drawbacks with capillary zone electrophoresis (CZE), the mode of operation employed for the assay of elastolytic activity was micellar electrokinetic chromatography (MEKC). Using a background electrolyte consisting of 35 mM sodium tetraborate, pH 9.3, containing 65 mM SDS and 15% v/v methanol, separation of intact peptide substrate from products of proteolytic reaction was easily achieved in a fused‐silica capillary of 50 cm effective length × 75 μm ID. This allowed us to determine the rate of hydrolysis of substrates and to calculate the kinetic parameters Km and kcat of the proteases investigated. A comparison of these data with those obtained from high performance liquid chromatography (HPLC)‐based experiments showed that MEKC is a convenient technique for studying protease kinetics.
Chiral separation of α‐amino acids by ligand‐exchange capillary electrophoresis using N‐(2‐hydroxy‐octyl)‐L‐4‐hydroxyproline as a selector
Authors: Végvári Ákos , Kilár Ferenc , Schmid Martin G. , Gübitz Gerald 
Published: May 2005

AbstractThe direct chiral resolution of underivatized α‐amino acids by capillary zone electrophoresis (CZE) based on the principle of ligand exchange is described. An N‐(2‐hydroxyoctyl)‐L‐4‐hydroxyproline/Cu(II) complex was used as a chiral selector. Besides amino acids containing aromatic residues, the basic amino acid histidine was resolved. Baseline separations were obtained for all amino acids investigated. The influence of selector concentration, electrolyte composition and pH on the resolution was investigated. It was found that there is a correlation between pI of the amino acids and the optimal pH.
Nonaqueous capillary electrophoresis — its applicability in the analysis of food, pharmaceuticals and biological fluids
Authors: Bjørnsdottir Inga , Tjørnelund Jette , Hansen Steen Honoré 
Published: May 2005

AbstractThe use of nonaqueous electrophoresis media for the application of capillary electrophoresis in the analysis of food, pharmaceuticals and biological fluids is reviewed. Some of the applications are discussed in detail and the benefits of using nonaqueous media in these cases are outlined. Three new applications within pharmaceutical analyses are presented. In these methods either a simple sample pretreatment by dilution with methanol (determination of chlorhexidine in a cream) or selective on‐line capillary electrophoresis mass spectrometry (methods for identification of seizure drugs or opium alkaloids) are used. The choice of organic solvents and electrolytes for nonaqueous capillary electrophoresis are discussed. Furthermore, validation data obtained using capillary electrophoresis based on the nonaqueous principle are listed and discussed.
Nonaqueous capillary zone electrophoresis for separation of free fatty acids with indirect fluorescence detection
Authors: Wang Tianlin , Wei Hongping , Li Sam F. Y. 
Published: May 2005

AbstractThe feasibility of combining nonaqueous capillary zone electrophoresis with indirect fluorescence detection was studied for the efficient separation and sensitive detection of ionizable hydrophobic substances which do not possess practically suitable detection properties. Medium‐ and long‐chain free fatty acids, C6C24, were selected as test compounds. The results showed that such a wide range of medium‐ and long‐chain free fatty acids could be separated between any two consecutive homologs in one run and be detected at a level of about 0.01–0.02 mM in highly basic methanol/acetonitrile media containing fluorescein as coion of background electrolyte for indirect fluorescence detection.
Electrochemical detection in capillary electrophoresis with dual‐parallel on‐capillary electrodes
Authors: Voegel Phillip D. , Baldwin Richard P. 
Published: May 2005

AbstractA new approach for dual electrode electrochemical detection in capillary electrophoresis (CEEC) is described. In this approach, two identical capillaries, each containing an on‐capillary electrode incorporated permanently onto its tip, were paired together for simultaneous sample injection and detection. This procedure permitted dual‐parallel detection to be performed without the need for painstaking alignment of the electrodes with respect to one another and to the capillary outlet as is required for the off‐capillary microelectrode systems usually employed in CEEC. As a result, independent detection at two electrodes held at different potentials or at two electrodes of different composition or structure could be performed simply and with wide flexibility. Fabrication of on‐capillary electrodes was carried out by sputter‐coating the exit end of the capillaries with a thin layer of Au or Pt. Dual electrode system performance was demonstrated by separation and analysis of phenol and catechol samples. In addition, the detection system was coupled with glucose oxidase for the selective detection of glucose.
Cyclodextrins as selectivity enhancers in capillary zone electrophoresis of proteins
Authors: Rathore Anurag S. , Horváth Csaba 
Published: May 2005

AbstractThe selectivity in the capillary zone electrophoresis (CZE) of a variety of acidic and basic proteins including α‐chymotrypsinogen A, cytochrome c, lysozyme, ribonuclease A, ovalbumin, and β‐lactoglobulins A and B, was altered by adding 6‐monodeoxy‐6‐monoamino‐β‐cyclodextrin or carboxymethylated β‐cyclodextrin to the electrophoretic medium of aqueous 50 mM sodium phosphate, pH 2.5. On the other hand, no significant improvement was obtained in the separation upon addition of heptakis (2,6‐di‐O‐methyl)‐β‐cyclodextrin. Whereas protein adsorption on the wall of raw silica capillaries was significant in the absence of cyclodextrin, by addition of β‐cyclodextrin or its derivatives to the background electrolyte, wall adsorption was reduced with concomitant enhancement of the recovery. The results confirm that in various separation techniques, particularly those which employ microcolumns, certain cyclodextrin additives can be useful selectivity enhancers not only in the separation of small sample molecules but also in that of proteins.
Assay of trypsin activity by capillary isoelectric focusing with laser‐induced fluorescence detection
Authors: Shimura Kiyohito , Kasai Ken‐ichi , Matsumoto Hiroyuki 
Published: May 2005

AbstractCapillary isoelectric focusing is a highly effective method for the separation of proteins due to focusing as a function of their pI values in the separation process. This technique is also effective for certain types of peptides that focus well. Fluorescence labeling and subsequent detection by laser‐induced fluorescence farther enhance the sensitivity of this technique. This paper demonstrates the utility of this technique in an enzyme assay. A synthetic nona peptide, H‐Gly‐Cys‐His‐Glu‐Ala‐Arg‐Ala‐Glu‐Glu‐OH, was labeled with an iodoacetyl derivative of Lissamine rhodamine B at the thiol group of the cysteine residue as a substrate for trypsin. Trypsin catalyzed the cleavage of the Arg‐Ala bond of the labeled substrate, which focused at pH 4.8, and liberated a shortened, labeled product, H‐Gly‐*Cys‐His‐Glu‐Ala‐Arg‐OH that focused at pH 6.9 (*indicates the label). The product peptide at 3–300 pM was determined with a relative standard deviation of 5.5% (n = 5) by fluorescence detection at 590 nm with excitation by a green line of He‐Ne laser. Incubation of trypsin with the substrate for 10 min at 37°C allowed the determination of 50–250 pg of trypsin, with a relative standard deviation of 5.3% (n = 5).
Protein profile characterization of bacterial lysates by capillary electrophoresis
Authors: Kustos Ildikó , Kocsis Béla , Kerepesi Ildikó , Kilár Ferenc 
Published: May 2005

AbstractA fast and reproducible method was developed to characterize cell lysates by their electrophoretic profiles using capillary electrophoresis (CE). Characteristic and reproducible patterns were recorded for each bacterial strains when “dynamic sieving” CE, using a polymer solution in the capillary, was applied to distinguish four strains of the Enterobacteriaceae family. The electropherograms showed distinct differences when comparing them to the sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) protein profiles. This is certainly a result of the differences in the separation principles and in the detection methods of the two techniques.
A comparison of liver protein changes in mice and hamsters treated with the peroxisome proliferator Wy‐14,643
Authors: Giometti Carol S. , Tollaksen Sandra L. , Liang Xiaoli , Cunningham Michael L. 
Published: May 2005

AbstractInterspecies differences in the liver response to Wy‐14,643, a potent peroxisome proliferator in rats and mice, have been demonstrated. While both rats and mice show dramatic increases in the number of peroxisomes, the activity of peroxisomal enzymes involved in the β‐oxidation of fatty acids, and heptocyte replication, Syrian hamsters have a more moderate peroxisome proliferation response and no sustained increase in cell replication. Rats and mice, but not hamsters, develop hepatocellular carcinoma after prolonged exposure to Wy‐14,643. To further characterize this species difference, two‐dimensional gel electrophoresis (2‐DE) has been used to compare the effect of 14‐day exposure to various dietary concentrations of Wy‐14,643 on liver protein expression in male mice and hamsters. Digitized images of the 2‐DE protein maps were searched for significant changes. The peroxisome bifunctional enzyme (PBE) enoyl CoA hydratase/3‐hydroxyacyl dehydrogenase, which migrates to the same position in mouse and hamster liver protein 2‐DE patterns, increased in abundance by more than three times the control level in both mice and hamsters. In addition to the quantitative change in PBE, significant quantitative changes (P < 0.001) were found in 49 mouse liver proteins (47 decreasing and 2 increasing) and in 35 hamster liver proteins (27 decreasing and 8 increasing). There was little overlap in the mouse and hamster proteins showing quantitative changes in response to Wy‐14,643, with the exception of PBE and one unidentified liver protein with an approximate molecular weight of 50 000. These results show that although peroxisome proliferation occurs in the livers of both mice and hamsters exposed to Wy‐14,643, other species‐specific changes in proteins occur that are independent of the peroxisome proliferation response and that could be related to species‐specific susceptibility or resistance to liver tumor induction.
Characterization of sugar chain structures of human α‐fetoprotein by lectin affinity electrophoresis
Authors: Taketa Kazuhisa 
Published: May 2005

Abstractα‐Fetoprotein (AFP) glycoforms, defined as AFP with different chemical structures of carbohydrate, were analyzed by affinity electrophoresis with several lectins of known specificities against complex‐type oligosaccharides. Serum AFP samples from cord blood on full‐term delivery and from patients with hepatocellular carcinoma and extrahepatic malignancies including gastrointestinal tumors and yolk sac tumors were used. Two‐dimensional lectin affinity electrophoresis and also lectin affinity chromatography coupled with lectin affinity electrophoresis were employed. More than ten AFP glycoforms were identified or characterized using the above‐mentioned AFP samples. Known specificities of the lectins against complex‐type oligosaccharides were refined or their additional specificities were found in this study. Lectin appeared to have specificity against carbohydrates by recognizing not only specific residues but also the whole carbohydrate molecule containing the residues, resulting in differential affinities for the lectin.
Analysis of starch structure using fluorophore‐assisted carbohydrate electrophoresis
Authors: Morell Mathew K. , Samuel Michael S. , O'Shea Michael G. 
Published: May 2005

AbstractThe analysis of the fine structure of starches is important to the investigation of linkages between starch structure and function and to the investigation of the properties and roles of starch biosynthetic, modifying and degradation enzymes. Fluorophore‐assisted carbohydrate electrophoresis has recently been introduced as a method for the analysis of the oligosaccharide populations released by the enzymatic digestion of starches, which has advantages in resolution and sensitivity over previously used methods, and provides the capacity for the facile analysis of oligosaccharide populations on either a molar or mass basis. The use of fluorophore‐assisted carbohydrate electrophoresis for the analysis of oligosaccharides is reviewed with particular reference to the choice of label, efficiency of labeling and separation techniques. Examples of separations using slab gel electrophoresis, DNA sequencer analysis and capillary electrophoresis are presented and we conclude that on the basis of resolution and reproducibility, capillary electrophoresis is the method of choice for the separation of oligosaccharides of degree of polymerization from 1 to 100. Examples of isoamylase‐debranched starches and glycogens analyzed by capillary electrophoresis are presented. The capillary electrophoresis analysis of starch structure through the analysis of oligosaccharides released by the debranching of limit dextrins derived from starches and glycogens is introduced as a useful diagnostic of starch structure. The potential for future development of novel diagnostics for starch structure using fluorophore‐assisted carbohydrate electrophoresis is discussed.
Preparative separaton of plasmid and bacterial artificial chromosome DNA by density gradient electrophoresis in the presence of linear polymers
Authors: Cole Kenneth D. 
Published: May 2005

AbstractA density gradient apparatus was used to examine the separation of different physical forms and sizes of DNA. A gradient of sucrose was used to stabilize thermal convection during electrophoresis in the column (2.2 cm in diameter). Linear polymers were added to the density gradient and screened for their ability to separate the supercoiled, nicked circular, and linear forms of the plasmid pBR 322. The influence of different concentrations and molecular weights of the polymers was examined on the separation. Polyethylene oxide with a molecular weight of 5 000 000 and a concentration of 0.2% w/v achieved the best separation results for the different physical forms of the plasmid. The order of separation of the different physical forms of the plasmid were linear (fastest), supercoiled, and nicked circular (slowest). These conditions were also used to separate a preparation of bacterial artificial chromosome (BAC) DNA. A rapidly moving form, presumably the supercoiled form, was resolved from a large amount of E. coli genomic DNA and from sheared forms of the BAC DNA.
Capillary electrophoretic separation of 1 to 10 kbp sized dsDNA using poly(ethylene oxide) solutions in the presence of electroosmotic counterflow
Authors: Chen Hsuan‐Shen , Chang Huan‐Tsung 
Published: May 2005

AbstractDNA fragments of 1 to 10 kbp in length were separated by capillary electrophoresis (CE), using poly(ethylene oxide) (PEO) solutions in the presence of electroosmotic flow. The technique requires filling the capillary with the polymer solution by means of electroosmotic flow (EOF). Separation times of 6–7 min in PEO solutions ranging from 0.3 to 8 × 106 Mr at 375 V/cm were sufficient to separate the 11 components of the dsDNA ladder (0.5 to 10 kbp) by size. The migration behavior of the double‐stranded (ds)DNA fragments, interpreted by “Ferguson plot analysis”, in the system is indistinguishable from that previously reported for capillary zone electrophoresis (CZE) in a polyacrylamide solution without EOF. Potential advantages of conducting CZE using polymer solutions in the presence of EOF are: (i) Possibility of long migration times on short columns; (ii) possibility of introducing relatively viscous, high Mr polymer solutions into narrow capillaries; (iii) possibility of establishing polymer concentration gradients in capillaries; (iv) possibility of concentrating the starting zone by balancing electrophoretic migration and electroosmotic transport.
Thermal degradation and flammability of cellulosics
Authors: Košík M. , Luzáková V. , Reiser V. , Blažej A. 
Published: May 2005

AbstractThe course of pyrolysis, thermooxidation, flame combustion and glowing combustion has been observed with differently treated cellulose samples by means of thermoanalytical methods. The impregnation of cellulose materials by inorganic salts produces a less combustible material, less stable at relatively low temperatures, yet giving more of the solid residue resistant to pyrolysis and combustion processes. The more thermally stable cellulose derivatives, but with insufficient resistance to flame and particularly glowing combustion, have been obtained by cyanoethylation and grafting of acrylonitrile on to cellulose.
Flame‐resistant fibres and fabrics a review of the work at lambeg industrial research association
Authors: Hill B. J. 
Published: May 2005

AbstractThis paper discusses the relative merits of a number of flame‐resistant fabrics. The information quoted draws heavily from a five year programme of research carried out on the subject by Lambeg Industrial Research Association. The fabrics concerned were made either from flame‐resistant fibres or by flame‐resistant treatment of fabrics. The textile problems which can arise in the production of these fabrics are considered and means of avoiding them suggested. Test methods for flammability are discussed briefly and the subject of toxic gases generated from flame‐resistant materials in fire situations is introduced as an area which needs further study. The paper is essentially a situation report describing the present state of knowledge indicating gaps therein, the limitations of the fibres, fabrics and finishes available and hence areas for future work.
Measurement of smoke in fires—a review. I —characteristics of smoke
Authors: Maries K. 
Published: May 2005

AbstractThe Complexity of the behavior of smoke at fires can lead to great difficulties in obtaining measurements from instruments which are both reliable and correlatable with human observations. The first article in this two‐part review considers the principal characteristics of smoke, the factors which influence its behaviour at fires, the basic theory of light transmission through smoke, and the behaviour of the eye in smoky conditions. This information will be used in the second article to assess the performance and limitations of various types of smoke measuring instruments.
The effect of the carbon‐bromine bond type on the fire retardation of unsaturated polyester
Authors: Ravey M. , Fischler Th. , Kramer M. 
Published: May 2005

AbstractLimiting Oxygen Indexes were determined for a polyester resin fire retarded with a number of different brominated hydrocarbons. It was found that up to a concentration of 28 phr bromine the values obtained were linearly related to the bromine concentration and independent of the carbon‐bromine bond.