bluuebunny/crossref_metadata_2025_split huggingface dataset:
Separation of different forms of transferrin by isoelectric focusing to detect effects on the liver caused by xenobiotics
Authors: Petrén Sven , Vesterberg Olof 
Published: May 2005

AbstractSeveral genetic variants and also isoforms of transferrin differing in carbohydrate structure can be separated by polyacrylamide or agarose gel isoelectric focusing. Numerous blood plasma or serum samples can be analyzed in parallel in each gel. Studies of the heterogeneity of transferrin have already revealed many results of importance to different fields of human medicine. Gene typing can give important and useful information for paternity determination and in forensic medicine. The gene type C 2 seems to have increased frequency in certain malfunctions. Furthermore, functional abnormalities of liver cells can be revealed by determination of the concentrations of transferrin isoforms differing mainly in their carbohydrate parts. The isoforms can be quantified with zone immunoelectrophoresis assay. Thus valuable information can be obtained about important modulated regulations of cell and membrane functions, even when these are disturbed by disease and xenobiotics. The information may be usefule.g. in the detection of individuals suffering from toxic effects, to identify toxic agents and exposure conditions. Studies of house painters revealed that exposure to different types of paints had an effect on transferrin. Determination of the concentration of the isotransferrin with pI5.7 in blood samples from alcoholics can be used as a marker for the detection of liver dysfunction and for the monitoring of therapy treatments. In addition, by analyzing the isotransferrins a rare genetic abnormality can be detected.
Electrophoretic studies of antimitochondrial antibodies: Identification of mitochondrial antigens specific to primary biliary cirrhosis
Authors: Ishii Hideo , Muno Daisaku , Namihisa Toshihiko 
Published: May 2005

AbstractMitochondrial inner membrane proteins extracted from beef heart tissue were examined for reactivity to antimitochondrial antibody (AMA) present in sera of patients with primary biliary cirrhosis (PBC) by an immunoblotting technique. Four proteins, which reacted with AMA, had molecular masses of 70 kDa, 50 kDa, 47 kDa and 40 kDa, as defined by their relative mobility (Rf) in sodium dodecyl sulfate‐polyacrylamide gel electrophoresis. All sera of 114 PBC patients were positive with at least one and as many as four of the mitochondrial proteins. The major antigenic proteins of mitochondrial inner membrane to which AMA reacts were the 70 kDa and 47 kDa proteins. All PBC sera containing antibodies to the 50 kDa and/or 40 kDa proteins reacted with 70 kDa as well. The isolation of antigen reacting with AMA of PBC is important to warrant further study of AMA and the cause of the disease. The isolation of responsible antigens had been difficult because the four antigens were insoluble. However, the antigen newly found by us, the 36 kDa fragment, obtained by partial trypsin digestion, is soluble. Using several procedures, the antigenic protein target of AMA was purified from mitochondria for the first time. We determined the N‐terminal sequence of the soluble 36 kDa fragment, 25 residues in length. Until now the N‐terminal sequence of the 36 kDa protein has not shown significant homology with any known protein. The present results of antigen purification would contribute to the elucidation of the epitopes of AMA antigen.
Antigen‐specific electrophoretic cell separation for immunological investigations
Authors: Hansen Ernil , Wustrow Thomas P. U. , Hannig Kurt 
Published: May 2005

AbstractPreincubation of human blood lymphocytes with cell surface antigen specific antibodies under non‐capping conditions reduces the electrophoretic mobility of the corresponding lymphocyte subpopulation. Antigen‐positive and antigen‐negative cells can be separated by free flow electrophoresis with high yield, purity and viability. The use of fluorescence‐labelled second antibodies augments the induced decrease in net surface charge density, and allows rapid detection of antigen‐positive cells in the fractions of electrophoresis. Carrier‐free cell electrophoresis of human peripheral blood lymphocytes after reaction with anti‐IgM‐antibody or the monoclonal antibodies OKT4 or OKT8, and sandwich staining with tetrarhodamine isothiocyanate‐labelled anti‐IgG resulted in the large‐scale separation of highly pure human B and T lymphocyte subpopulations. Their functional integrity was shown in assays of lymphocyte transformation and of antigen‐specific induction and regulation of antibody synthesis in vitro. These separated lymphocyte subpopulations are useful tools for immunological investigations. While, for instance, the effects of drugs on human lymphocytes are obscured by coincident changes in cell composition of the peripheral blood tested that do not by themselves reflect whole body immunocompetence, the cell separation and in vitro assays at a defined cell number and cell composition allow the recording of quantitative changes in the function of different cell sub‐populations. We studied the influence of the anesthetic thiopental on separated, human lymphocyte subsets. In both polyclonal lectin stimulation and in vitro antibody production, thiopental exhibited a noncytotoxic suppression of lymphocyte functions. B‐Cells, T‐helper and T‐suppressor cells were equally affected and showed the same dose response. The electrophoretic separation of cells after reaction with antibodies is applicable to a wide range of human lymphocyte subpopulations defined by specific antibodies. Their gentle isolation in large quantities is a powerful tool for investigations of the immune system.
Polymorphism of aldehyde dehydrogenase and its application to alcoholism
Authors: Harada Shoji 
Published: May 2005

AbstractAldehyde dehydrogenase (ALDH) consists of four different isozymes (I, II, III, IV). Among these, ALDH I shows genetic polymorphism (normal and deficient) in a population of Mongoloid origin. The significantly lower frequency of ALDH I deficiency was found in alcoholic patients compared with healthy controls. ALDH I deficiency is one of the important genetic factors in regulating alcohol consumption and plays a protective role against alcoholism. Population genetic studies on ALDH I deficiency in different ethnic groups indicate that ALDH deficiency was found only in Mongoloid population groups.
Characterization of bacterial genospecies by computer‐assisted statistical analysis of enzyme electrophoretic data
Authors: Picard Bertrand , Goullet Philippe , Bouvet Philippe J. M. , Decoux Gerard , Denis Jean‐Baptist 
Published: May 2005

AbstractA computer‐assisted statistical treatment of the electrophoretic data obtained from the analysis of two dehydrogenases and 27 kinds of esterases produced by strains belonging to the taxonomically complex genus Acinetobacter is described. The 12 genospecies were clearly separated from each other by correspondence analysis, For each genospecies the distances of the strains from their barycenter were computed and typical isolates suitable for use as reference strains were determined. This approach is suitable for the systematic study of other procaryotic or eucaryotic organisms.
Electrophoresis in one buffer at two pH values
Authors: Michov Budin M. 
Published: May 2005

AbstractA theory for discontinuous electrophoresis in a polyacrylamide gel is presented for one buffer at two pH values. It is shown that polyions stack between identical leading and trailing ions, and resolve in a gel of constant polyacrylamide concentration. The theory is illustrated by the separation of serum protein polyions in a Tris‐glycinate buffer of pH 8.19 in the well‐forming gel, and pH 9.16 in the resolving gel. The selected concentrations and electrolyte ionization degrees of Tris and glycine have values at which the serum protein polyions stack between the resolving and electrode buffers, followed by separation in the resolving gel.
Determination of the electrophoretic mobility of chromosomes by free flow electrophoresis: I. Morphology and stability
Authors: Bier Frank F. , Bettag Ulrich , Rheingans Thomas , Adrian Hedwig , Barths Jochen , Hausmann Michael , Bühring Hans‐Jörg , Rohwer Peter , Dölle Jürgen , Cremer Christoph 
Published: May 2005

AbstractIsolated metaphase chromosomes of several fibroblastoid cell lines (Chinese hamster, Chinese hamster × human hybrid) were subjected to free flow electrophoresis (FFE) to study their electrophoretic mobility (EM). The morphology and stability of the chromosomes were unaffected by FFE as examined by cytogenetic methods and flow cytometry. The chromosomes of the complement all showed similar EM under most of the conditions applied. At neutral pH the EM of the chromosomes had the same sign as free DNA and about 2/3 of its magnitude. The variation of EM with buffer parameters such as ionic strength, valence of counterions, buffer capacity and dielectric constant of the solvent were investigated. Thermal denaturation increased the EM of the chromosomes by 20 %. Partial denaturation might offer a possibility to separate or enrich large amounts of chromosomes by FFE.
Gel electrophoresis of native actin and the actin‐deoxyribonuclease I complex
Authors: Edgar Alasdair J. 
Published: May 2005

AbstractElectrophoresis of monomeric actin (G‐actin) on 8–25 % acrylamide Pharmacia PhastGels was carried out using gels and agarose buffer strips preequilibrated in buffer containing adenosine triphosphate (ATP), calcium ions (Ca2+) and dithiothreitol. On these gels G‐actin ran as a sharp band at an apparent molecular mass of 45 kDa relative to standard proteins which is slightly greater than its actual molecular mass of 42 kDa. Electrophoresis in the absence of these solutes led to denaturation and aggregation of the protein, as reflected by a long streak. Filamentous actin (F‐actin) did not enter the gel. The actin monomer‐binding protein, deoxyribonuclease I, (DNase I) forms a binary complex with G‐actin. The purity and apparent molecular mass 74 kDa of this complex were determined by native gel electrophoresis. By the simple procedure of preequilibrating both gel and buffer strips with appropriate ligands, this technique could be extended to investigate interactions between actin and other G‐actin‐binding proteins and other proteins whose stability is ligand dependent.
Rapid and sensitive silver‐lipopolysaccharide staining using PhastSystem in fast horizontal polyacrylamide gel electrophoresis
Authors: Lee Kuo‐Kau , Ellis Anthony E. 
Published: May 2005

AbstractA rapid and sensitive silver‐lipopolysaccharide staining method has been developed by using PhastSystem. The total time of the procedure (including time of Phastgel electrophoresis) is within 2 h. It is at least 10 times faster than the previous reported methods and the sensitivity is also increased.
The use of PhastSystem crossed immunoelectrophoresis with immunoblotting to demonstrate a complex between glycoprotein Ib and the actin‐binding protein (ABP) of human platelets
Authors: Aakhus Anne‐Margrethe , Wilkinson Michael , Pedersen Turid Margrethe , Solum Nils Olav 
Published: May 2005

AbstractThe study shows how a technique described in an accompanying paper can be applied to solve a biological problem. The technique makes use of the observation that a monoclonal antibody that has been coprecipitated with its antigen during crossed immunoelectrophoresis can be transferred to a nitrocellulose membrane and visualized. Previous studies using crossed immunoelectrophoresis of Triton X‐100 extracts of platelets have indicated that a particular immunoprecipitate (peak III) of the membrane receptor glycoprotein Ib (GP Ib) might contain a complex between the receptor and the actin‐binding protein (filamin). When a monoclonal antibody (PM6/ 317) directed towards the actin‐binding protein was added to a platelet extract prior to immunoelectrophoresis and blotting, this was visualized on the blot as a replica of the peak III immunoprecipitate. This demonstrates a colocalization of GP Ib and the actin‐binding protein in the precipitate, and thus the existence of a complex between the membrane receptor and the cytoskeletal protein.
Polyacrylamide gradient gel electrophoresis of cytosolic aretinol‐ and retinoic acid‐binding proteins: Application to rat testis and liver
Authors: Melin Anne‐Marie , Carbonneau Marie‐Annette , Maviel Marie‐Josée , Clerc Michel 
Published: May 2005

AbstractDistribution and cellular levels of retinol‐binding protein and retinoic acid‐binding protein, involved in the molecular action of retinoids, were analyzed in rat testis and liver. Both binding proteins of cytosolic extracts were separated by linear‐polyacryl‐amide gradient gel electrophoresis and following electrophoretic separation, could be visualized by complementary identification tests such as autoradiography and marker proteins. The concentrations of the binding proteins were evaluated by scanning the polyacrylamide gradient gels and the resulting data were found to be in accordance with those obtained by counting radioactivities. Polyacrylamide gradient gel electrophoresis appears suitable to detect and quantitatively evaluate cytosolic retinol‐ and retinoic acid‐binding proteins.
Two‐dimensional electrophoresis of plasma proteins and high density lipoproteins during inflammation
Authors: Choukaife Ala , Visvikis Sophia , Steinmetz Josiane , Galteau Marie‐Madeleine , Kabbaj Ouafae , Férard Georges , Métais Pierre , Siest Gérard 
Published: May 2005

AbstractPlasma protein and lipoprotein fractions of five patients were analyzed on day 1, 5, and 15 after severe head injury by combining three types of two‐dimensional electrophoresis (2‐DE) to obtain information on lipoprotein and apolipoprotein composition. On analysis under nondenaturing conditions in both dimensions on day 5, the samples show modifications of isoelectric point (pI) and molecular weight (Mr) properties of the high density lipoprotein (HDL) fraction in addition to an increase in inflammatory proteins and a return to a normal pattern on day 15. In the second type of 2‐DE the samples were analyzed employing isoelectric focusing without denaturant in the first dimension, followed by sodium dodecyl sulfate (SDS) in the second dimension in order to study the protein composition of lipoprotein fractions. On day 5, a decrease of the apolipoproteins apo A‐I, apo A‐II, and apo C were noted, with simultaneous appearance of an unidentified protein with Mr 12 000 and pI 6.0. In the third type of 2‐DE, employing urea and Nonidet P‐40 in the first and SDS in the second dimension, the plasma polypeptide composition was studied. The presence of an unidentified polypeptide could be confirmed on day 5, tending to disappear there‐after. This Mr 12 000 component consists of two major spots at pI 5.7 and 6.0 and four minor ones between pI 6.0 and 8.0. These properties suggest that this protein corresponds to serum amyloid A apolipoprotein.
Analysis of virus protein heterogeneity among group B coxsackie viruses using a “mini” two‐dimensional gel electrophoresis system
Authors: Cash Phillip 
Published: May 2005

AbstractThe application of a small format two‐dimensional polyacrylamide gel electrophoresis (2D‐PAGE) system to the study of protein heterogeneity among group B coxsackie virus (CVB) isolates is described. Under the conditions of electrophoresis developed during this study, protein samples could be processed within 7 h and up to 300 intracellular proteins were resolved from uninfected HEp‐2 cell lysates. 2D‐PAGE was used to characterise the intracellular proteins of clinical CVB isolates of serotypes 4 and 5. Intracellular proteins from virus‐infected cells were radiolabelled using a pulse‐chase protocol under conditions which promoted inhibition of cellular protein synthesis. Depending on the CVB serotype up to 11 intracellular virus proteins were identified, ranging in molecular weight between 14 000 and 54 000. Although the overall two‐dimensional protein profiles were characteristic for the two CVB serotypes, within a CVB serotype there was some heterogeneity of the virus proteins, mainly affecting the proteins' net charge. The sensitivity of 2D‐PAGE in detecting subtle differences in virus proteins combined with the convenience of the small gel format makes this a suitable approach for the study of the molecular epidemiology of human virus pathogens.
Thin‐layer electrophoretic separation of monosaccharides, oligosaccharides and related compounds on reverse phase silica gel
Authors: Scherz Heimo 
Published: May 2005

AbstractThe electrophoretic mobilities of monosaccharides, oligosaccharides, sugar alcoho and sugar acids were determined in 0.3 M borate buffer, pH 10, using thin‐layer electrophoresis on silanized silica gel, pretreated with octanol‐1. A rapid separation of number of sugars, occurring in foods, could be achieved. Using a 0.05–0.1 M neutra solution of barium acetate as electrolyte, thin‐layer electrophoresis allowed exceller and rapid separation as well as identification of all common uronic acids which are constituents of many acidic polysaccharides.
Isolation and recovery of acidic oligosaccharides from polyacrylamide gels by semi‐dry electrotransfer
Authors: Al‐Hakim Ali , Linhardt Robert J. 
Published: May 2005

AbstractAcidic oligosaccharides derived from glycosaminoglycan heparin were separated by polyacrylamide gradient gel electrophoresis (PAGE). The gel could be visualized using Alcian Blue dye to give a pattern of highly resolved, well defined bands. The particular banding pattern obtained was the result of a heparinase catalyzed depolymerization which afforded oligosaccharide products that differed in size by one disaccharide unit. The separated oligosaccharides could be recovered prior to staining by electroelution onto a positively charged nylon membrane by a semi‐dry transfer procedure. Subsequent elution and quantitative recovery of individual oligosaccharides from the membrane was achieved. By using multiple membrane layers a second separation dimension was obtained, resulting in increased oligosaccharide purity proportional to transfer depth. Preparative gradient polyacrylamide gel electrophoresis followed by semi‐dry electro‐transfer and recovery represents a novel method for the preparation of homogeneous acidic oligosaccharides.
Multiple successive immunoprinting: A fast blotting technique of a single agarose isoelectric focusing gel
Authors: Desvaux François X. , David Bernard , Peltre Gabriel 
Published: May 2005

AbstractMultiple successive pressure blottings of a single agarose isoelectric focusing gel were performed on normal and CNBr‐activated nitrocellulose (NC) filters. The results obtained by multiple successive 10s immunoprints were compared to those obtained by a single 10 min immunoprint. To quantify the transfer efficiency of these techniques, a defined amount of radioactive material was separated by isoelectric focusing on agarose gel. After separation and pressure blottings of the gel the NC filters were submitted to autoradiography. The amount of radioactive material bound to the NC filters was determined by scintillation technique. The single 10 min pressure blot was more efficient than each of the multiple successive 10s prints. However, the latter procedure allowed equal resolution and resulted in a higher recovery of total radioactivity than the single immunoprint technique. The aim of this paper is to show how to obtain highly reproducible prints of electrophoretic patterns in an agarose gel of heterogeneous samples when accurate multiple immunodetections are to be performed. This technique was tested to characterize the grass pollen specific immunoglobulin classes and subclasses from an allergic patient.
Analysis of plasma membrane protein changes in Dictyostelium discoideum during concanavalin A induced receptor redistribution using two‐dimensional gel electrophoresis
Authors: Patton Wayne F. , Dhanak Michelle R. , Jacobson Bruce S. 
Published: May 2005

AbstractThe 127 major polypeptides obtained from the purified plasma membrane of Dictyostelium discoideum were examined using two‐dimensional gel electrophoresis and a microcomputer‐based videodensitometer. Plasma membrane proteins were analyzed at four discrete stages of concanavalin A induced cell surface capping; (i) the cell surface in the absence of ligand (unbound), (ii) the surface immediately after ligand binding (bound), (iii) the cell surface after receptors had patched (patched) and (iv) the cell surface after receptors had capped (capped). Plasma membranes were obtained at various stages of capping by using a colloidal silica density perturbation technique which immediately immobilized the proteins, preserving their lateral distribution in the bilayer during the isolation. Proteins were characterized with respect to post‐translational modification changes resulting from the capping process as well as changes in their association with the plasma membrane fraction. Posttranslational changes of plasma membrane proteins, such as phosphorylation, methylation and proteolytic cleavage, were not observed during the four stages of capping. Myosin heavy chain phosphorylation, however, decreased almost twofold during patching and capping. Actin, which is known to colocalize directly underneath capped receptors did not appear to be recruited to the cap from the cytoplasm.
Separation of some triazine herbicides and their solvolytic products by capillary zone electrophoresis
Authors: Foret František , Šustáček Vladimír , Boček Petr 
Published: May 2005

AbstractThe separation of triazine herbicides and their solvolytic products by capillary zone electrophoresis in mixed water‐ethanol background electrolytes is described, allowing the analysis of picomole amounts of a triazine mixture in less than 15 min.
Electrophoretic purification of the alpha and beta subunits of phosphorylase kinase and evidence in support of the deduced amino acid sequences
Authors: Crabb John W. , Harris William R. , Johnson Charles M. , Sotiroudis Theodore G. , Kuhn Carl C. , Heilmeyer Ludwig M. G. 
Published: May 2005

AbstractA simple, rapid sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) method is presented for isolating the α, α' and β subunits of rabbit muscle phosphorylase kinase. The SDS‐PAGE procedure can yield milligram amounts of α and β from a single preparative gel and also allows isolation of the α' isozyme free of α. Notably the method provides the purified subunits in a form amenable to structural analysis. Edman degradation of α and α' reveal identical NH2‐terminal structures. Amino acid analysis of the electrophoretically purified α and β subunits are in good agreement with their deduced primary structures. The amino acid sequence of 488 residues in α and 713 residues in β were determined by gas phase Edman degradation. The data support the recently deduced primary structures of α (Zander et al., Proc. Natl. Acad. Sci. USA 1988, 85, 2929–2933) and of β (Kilimann et al., Proc. Natl. Acad. Sci. USA, 1988, 85, 9381–9385).
IPGMAKER: A program for IBM‐compatible personal computers to create and test recipes for immobilized pH gradients
Authors: Altland Klaus 
Published: May 2005

AbstractThe program “IPGMAKER” is a computational aid for creating and testing recipes for near‐linear immobilized pH gradients. It was written for fast IBM personal computers (with a Type 80386 processor and 80387 coprocessor) and compatibles equipped with a VGA, EGA or Hercules (mono) graphics card. The program is limited to the use of up to 10 acids and/or bases, and to ranges spanning between pH 2 and 12. The resulting recipes are presented either as final concentrations in the 2 chambers of a mixing device for linear gradients or as volumes from 0.2 moles/L stock solutions adjusted to a user‐defined average buffering power. One of the subroutines determines the pH, gradient slope and buffering capacity at any location of the gradient and includes a facility to estimate the pI of proteins from the composition of their primary structure.