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C Kempf

Publications and source records attributed to C Kempf.

63 records · Page 4Linked to original sources
PubMed1982

Autoradiographic grain counting by fully automated image analysis.

A method for automated quantitative analysis of single label microautoradiographs applied to DNA repair synthesis is proposed. It uses image analysis of the microscopic field by a Leitz TAS and allows an automated selection of the nuclei to be analysed. The use of a grey level gradient algorithm provides a relatively large independence from external variables such as staining intensity, nuclei density in the field, stability of illumination etc. Random choice of the microscopic field by the computer directed stage displacement and automatic focusing ensures a high degree of objectivity and independence from human intervention. The coefficient of correlation between the automatic and the visual grain count is 0.97 in the range of visually countable grain densities, while the precision of measurement is about 10%. The speed of measurement is satisfactory (40 to 60 s for the analyse of a microscopic field) taking in account that only occasional supervision is needed.

Animals↗
PubMed1981

Interaction of phenylisothiocyanate with human erythrocyte band 3 protein. II. Topology of phenylisothiocyanate binding sites and influence of p-sulfophenylisothiocyanate on phenylisothiocyanate modification.

The two structurally related probes, the apolar phenylisothiocyanate and the polar, water-soluble p-sulfophenylisothiocyanate, were analysed for their topological interaction with human erythrocyte band 3 protein. Upon thermolytic and peptic digestion of labeled erythrocyte ghosts, the membrane-integrated segments of band 3 protein, the 17,000 and 10,000 dalton peptides, were isolated. At 2 mM initial label concentration, 90% of the hydrophobic probe phenylisothiocyanate was recovered in the 10,000 dalton peptide, the remaining amount of label being associated with the 17,000 dalton fragment. Pretreatment of the membranes with 5 mM p-sulfophenylisothiocyanate followed by labeling with 2 mM phenylisothiocyanate results in a consistent reduction in binding of phenylisothiocyanate by 1 mol/mol isolated band 3 protein. p-Sulfophenylisothiocyanate reportedly binds to the 17,000 dalton fragment (Drickamer, K. (1977), J. Biol. Chem. 252, 6909-6917). The interaction of the polar probe with the membrane protein affects binding of phenylisothiocyanate to the 10,000 dalton peptide by the equivalent of 1 mol/mol isolated peptide. The topological interrelation of the membrane-integrated segments is concluded.

Anion Exchange Protein 1, Erythrocyte↗
PubMed1980

Interaction of phenylisothiocyanate with human erythrocyte band 3 protein. I. Covalent modification and inhibition of phosphate transport.

The hydrophobic probe phenylisothiocyanate is utilized for chemical modification of human erythrocyte band 3 protein. The binding of phenylisothiocyanate to this protein is characterized in whole erythrocytes, erythrocyte ghost membranes and in isolated band 3 protein. The label, reactive with nucleophiles in their deprotonated form is found in all three preparations to be covalently bound to band 3 protein. Under saturation conditions, 4--5 mol phenylisothiocyanate are covalently bound per mol protein (molecular weight 95 000). The described modification effects inhibition of phosphate entry into erythrocytes. 50% inhibition of phosphate transport is obtained following a preincubation of erythrocytes with 0.45 mM phenylisothiocyanate. Both phenylisothiocyanate binding and transport inhibition are saturating processes. The relationship of the two parameters is non-linear.

Biological Transport↗
PubMedDate not supplied

Endogenous activation by indirect muta-carcinogens and DNA-repair synthesis in mouse-lung fibroblast cultures.

Cultures of adult mouse-lung fibroblasts have been treated by series of strong and weak muta-carcinogens. Unscheduled DNA synthesis has been measured by quantitative autoradiography using automatic image analysis. Some of the muta-carcinogens (AFB1, DMN, B(a)P, DMBA, MNNG, 4-NQO) yielded a measurable UDS response, whereas others (2-AA, AFB2, B(e)P, ICR-191) usually known as weak carcinogens, gave no response. The response was not improved by addition of liver S9. This shows that mouse-lung fibroblasts possess their own but limited metabolic activation systems.

Animals↗
PubMedDate not supplied

Strengths and limitations of the model virus concept.

New plasma- or cell culture-based pharmaceutical manufacturing processes must be validated for their ability to eliminate potentially contaminating pathogens. To evaluate the virus elimination potential of such a process, current guidelines propose the use of model viruses. This approach is discussed based on two examples. These examples show the strengths of this approach but also its limitations. The blood processing industry was recently challenged by the emergence of a West Nile Virus (WNV) epidemic in the United States. The susceptibility of WNV and a frequently used model virus to commonly used inactivation methods is compared. Current data show a good correlation. Due to its physico-chemical properties and the high viremic titers, B19 virus (B19V), a small (diameter 18-26 nm), robust, non-enveloped parvovirus, is a considerable challenge for the plasma processing industry. Mice minute virus (MMV), an animal parvovirus, is used as a model for B19V. Data show that B19V is considerably more susceptible to some physico-chemical inactivation methods than MMV. The examples of WNV and B19V show that the model virus concept is a practicable tool to evaluate the safety of plasma- or cell culture-derived pharmaceuticals regarding known and emerging viruses. It also underlines the need for investigational studies of relevant viruses if they can be handled in a normal virology laboratory, under moderate biosafety conditions.

Biopharmaceutics↗