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  • 1
    Electronic Resource
    Electronic Resource
    A three-dimensional solubility parameter approach to nonaqueous enzymology (1991)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 37 (1991), S. 627-638 
    Details
    ISSN: 0006-3592
    Keywords: solubility parameters ; hydrophobicity index ; Hansen parameter ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Widespread commercial application of enzymes as catalysts for specialty or commodity chemical synthesis will require their use in nonaqueous systems. While a number of non-aqueous enzyme applications have been demonstrated, the lack of useful rules for predicting enzyme-solvent interactions has hindered the development of this technology. Both Hildebrand and solvent hydrophobicity (octanol-water partition coefficient) parameters have been used previously to correlate and predict enzyme activity in nonaqueous systems, with some success, but any single-parameter approach is inherently limited in its ability to reflect the spectrum of possible enzyme-solvent interactions. Therefore, this study evaluates the three-dimensional solubility parameter space, as proposed by Hansen, to correlate and predict enzyme activity in microaqueous, miscible, and biphasic nonaqueous systems. Preliminary results suggest that Hansen parameters may be useful for correlating nonaqueous enzyme activity, and that the dispersive and polar parameters may be disproportionately important in single-phase microaqueous systems. The Hansen hydrogen-bonding parameter appears to be the only parameter yet evaluated capable of correlating the water requirement for enzyme activity in microaqueous systems, suggesting that water affects protein structure through enthalpic rather than entropic processes in nonaqueous systems. Insufficient data are available for miscible and biphasic systems, but it is proposed that enzyme activity may correlate with the average solubility parameters of miscible systems and of the aqueous phase in biphasic systems.
    Additional Material: 15 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260370705
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  • 2
    Electronic Resource
    Electronic Resource
    Balancing of energy-consuming processes of rat hepatocytes (1990)
    New York, NY [u.a.] : Wiley-Blackwell
    Cell Biochemistry and Function 8 (1990), S. 227-232 
    Details
    ISSN: 0263-6484
    Keywords: Energy-consuming processes ; rat hepatocytes ; oxygen consumption ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A method for the quantification of energy consuming processes described by Siems et al.6 for reticulocytes and by Müller et al.10 for ascites tumour cells was applied to balance the ATP-consumption of isolated rat hepatocytes. On the basis of decreased coupled respiration rates following the specific inhibition of energy-requiring reactions, the energy demands of protein turnover, nucleic acid synthesis, Na+/K+-ATPase and Ca2+-transport of hepatocytes in different incubation media were assessed. These processes together with urea synthesis account for about 60 per cent of the total energy consumption in a glucose and amino acid-enriched Eagle/Borsook medium. The metabolic flux rates of total ATP-consumption and ATP-consumption of single energy-requiring processes in hepatocytes are compared with those in reticulocytes and different tumour cell types.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/cbf.290080407
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  • 3
    Electronic Resource
    Electronic Resource
    Typing of the 3′ hypervariable region of the apolipoprotein B gene: Approaches, pitfalls, and applications (1993)
    Weinheim : Wiley-Blackwell
    Electrophoresis 14 (1993), S. 169-173 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Apolipoprotein B-100 is the principal protein component of lipoproteins with very low, intermediate, and low density. The interaction of apoB-100 with low density lipoprotein (LDL) receptors is responsible for the uptake of LDL into cells. An AT-rich hypervariable region is located adjacent to the 3′ end of the apoB gene. It consists of a variable number of tandemly repeated sequences (VNTR). Two approaches were used to analyze this polymorphism. In both, the region harboring the VNTR was amplified with the polymerase chain reaction (PCR). In the first method, fluorescently labeled primers were used in the PCR reactions and products were separated in agarose gels by means of an automated fluorescent fragment analyzer. In the second method, PCR products were analyzed in denaturing polyacrylamide gels and detected with silver staining. Even in the highly sophisticated automated system, agarose gel electrophoresis did not always enable unequivocal assignment of VNTR alleles. In contrast, denaturing polyacrylamide gel electrophoresis made it possible to distinguish the 15 bp differences between the VNTR alleles in a precise and simple manner. The VNTR polymorphism was typed in 234 individuals. Among these were 136 patients with coronary artery disease and 74 healthy controls. Thirteen alleles could be distinguished. The allele containing 49 repeats (VNTR-49) was found in 9.2% of the coronary artery disease patients and in 4.7% of the controls. Thus, the VNTR-49 allele increases relative coronary risk by about twofold. It is concluded that the apoB VNTR polymorphism is a potentially useful genetic marker. Since agarose gel electrophoresis may lead to ambiguous results, we prefer typing by denaturing polyacrylamide gel electrophoresis. This has to be accounted for, especially if the apoB VNTR polymorphism is applied to forensic studies.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150140128
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  • 4
    Electronic Resource
    Electronic Resource
    Clonal imbalances of plasma/serum immunogloblin production in infants (1993)
    Weinheim : Wiley-Blackwell
    Electrophoresis 14 (1993), S. 245-247 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: To study the clonal events occurring during ontogeny of the humoral immune system, we evaluated plasma immunoglobulin (Ig) production in term newborns and young children by high-resolution two-dimensional gel electrophoresis. The clonality pattern of Ig light (L) chains from healthy newborns (n = 19) was similar to that observed on protein maps of their mothers or of normal adults (n 〉 100), that is, rare distinguishable small spots in a cloud-like large band of indiscrete Ig L chain spots (polyclonal pattern; maternal Igs). Analysis of plasma samples obtained from infants between 1 month and 5 years of age (n = 55) revealed discrete but evident alterations of the clonality of Ig production. Between the 2nd and 4th months of life, transient attenuation of the “polyclonal background” was observed in association with the appearance of an increasing number of well-resolved Ig L chain spots (corresponding to plasma Ig concentrations between 0.5 and 1 g/L per spot). This “restricted” clonal pattern was progressivly less apparent on protein maps of infants older than 2 year and evolved towards a “normal” adult polyclonal pattern at the age of 5. These results suggest that the development of the B-cell clones is heterogeneous, either through limited outgrowth of precursor cells or through selective antigenic pressures.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150140142
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  • 5
    Electronic Resource
    Electronic Resource
    Pattern variations of polyclonal and monoclonal immunoglobulins of different isotypes analyzed by high-resolution two-dimensional electrophoresis (1993)
    Weinheim : Wiley-Blackwell
    Electrophoresis 14 (1993), S. 227-234 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: High-resolution two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) was used to analyze serum samples and purified immunoglobulins (Ig) obtained from “normal” individuals and from patients diagnosed with monoclonal gammopathies (MG) (n = 47; 5 IgA, 15 IgM, 15 IgG, 4 biclonal IgG, 1 IgD, 7 Bence Jones proteins). Polyclonal and monoclonal heavy (H) chains were located at different restricted gel positions according to their isotype. Monoclonal H chains appeared as sets of spots characterized by charge (pI) and size (Mr) microheterogeneity. Most of the monoclonal gamma chains were not seen on the gels (12/15). Supplementary polypeptides of 45-48 kDa were detected in serum samples containing monoclonal IgM, but were not seen in MG of other isotypes. However, these polypeptides were not specifically associated with monoclonal IgM because they were also found on protein maps of purified polyclonal IgM. Polyclonal light (L) chains appeared as cloudy bands containing several zones of higher density, whereas monoclonal L chains were usually resolved as single sharp spots. In 6 samples, monoclonal L chains were not seen, and in 9 samples, they appeared as two or more spots, characterized by different pI and/or Mr. In one sample obtained from a patient with a biclonal gammopathy, the L chains were resolved as 4 different spots. Our results confirm that 2-D PAGE is an excellent tool to study Ig. Analysis of the L chain region of the gels was particularly informative. Several monoclonal L chains exhibited heterogeneous two-dimensional spot patterns, suggesting that “subtile” clonal mutations of B-cell lineage and/or posttranslational modifications were involved in their production.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150140137
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  • 6
    Electronic Resource
    Electronic Resource
    Determining consanguinity by oligonucleotide fingerprinting with (GTG)5/(CAC)5 (1991)
    Weinheim : Wiley-Blackwell
    Electrophoresis 12 (1991), S. 397-402 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Simple tandemly organized (GTG)n/(CAC)n sequences are spread throughout the human chromosomes. The most informative DNA fingerprints for the testing of pedigrees and/or paternity were obtained with the simple triplet repeat probe (GTG)5 or its complement (CAC)5. These hypervariable simple-repeat fragments are stably inherited in a Mendelian fashion. Using these highly discriminating probes, all human individuals could, theoretically, be differentiated, except for genetically identical monozygotic twins. Examples from actual case work are reported and pertinent advantages of this methodology are discussed.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150120603
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  • 7
    Electronic Resource
    Electronic Resource
    Two-dimensional electrophoresis as an aid in the analysis of the clonality of immunoglobulins (1993)
    Weinheim : Wiley-Blackwell
    Electrophoresis 14 (1993), S. 1366-1371 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: In this paper, we summarize our five-year observation of two-dimensional polyacrylamide gel electrophoretic (2-D PAGE) analysis of immunoglobulin (Ig) light (L) chain patterns on serum/plasma and/or purified human Ig, and compare this technique with agarose electrophoresis and/or immunofixation examination. Polyclonal Ig L chains were seen as large “fuzzy” areas with several zones of high density. The majority (71%) of the monoclonal Ig L chains of monoclonal gammopathy detected by conventional electrophoresis appeared as a single large and well-defined spot on 2-D PAGE analysis, with the remaining appearing as multiple spots. The presence of oligoclonal Ig, reflected by multiple spots in 2-D PAGE, and several bands in immunofixation, was observed in 5 of 26 patients after allogeneic bone marrow transplantation and in 5 patients with tumors. In the majority (77%) of hypergamma-globulinemia, L chains appeared as a wide spread of small and well-defined spots in 2-D PAGE analysis. This pattern suggested oligoclonal Ig-secreting B cell clone expansion, and corresponding abnormalities were not detected with immunofixation. 2-D PAGE analysis also detected oligoclonal Ig expansions whereas conventional electrophoretic examination was normal in 10 additional patients after bone marrow transplantation, and in 6 of 10 immunocompetent patients with acute severe infections. Analysis of the Ig L chain pattern of a severe combined immune-deficient mouse populated with human peripheral blood leukocytes confirmed the skewed human Ig production in the model. In summary, 2-D PAGE appears to be a sensitive tool for the analysis of Ig diversity in various clinical situations.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.11501401210
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  • 8
    Electronic Resource
    Electronic Resource
    A large scale preparative gel electrophoresis separation of α1 and α2 subunits of the voltage-gated Ca2+ channel from rabbit skeletal muscle (1994)
    Weinheim : Wiley-Blackwell
    Electrophoresis 15 (1994), S. 1186-1190 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A large-scale preparative gel electrophoresis method effectively separates individual voltage-gated calcium channel (VGCC) subunits. with high resolution, starting with up to 4 mg of rabbit skeletal muscle L-type VGCC complex. Using this method, we separated α1 and α2 subunits of rabbit skeletal muscle VGCC with a high efficiency and with protein recoveries of 83%. The separated α1 and α2 subunits eluted from the gel in a 1:1 molar ratio. The method should be applicable for separating the other VGCC subunits or subunits of other protein complexes.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.11501501178
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  • 9
    Electronic Resource
    Electronic Resource
    Calculation of paternity probabilities from multilocus DNA profiles (1994)
    Weinheim : Wiley-Blackwell
    Electrophoresis 15 (1994), S. 170-174 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: We describe a procedure for evaluation of paternity evidence from multi-locus DNA probe patterns. A computer program abstracts a “+/-” notation description from the multilocus profile and then calculates a paternity index based on observed phenotypic fragment frequencies. The biostatistical evaluation considers only bands found in the child and missing from the mother - a simplified approach that is at once robust and conservative. Mutations are of course taken into account. Particular features lending objectivity to the interpretation include computer reading and matching decisions, and specific recognition and statistical compensation for ambiguities (“faint orphans”).
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150150129
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