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J Rickert

Publications and source records attributed to J Rickert.

4 recordsLinked to original sources
PubMed1998

Differential regulation of the clusterin gene by Ha-ras and c-myc oncogenes and during apoptosis.

Clusterin (ApoJ) is an extracellular glycoprotein expressed during processes of tissue differentiation and regression that involve programmed cell death (apoptosis). Increased clusterin expression has also been found in tumors, however, the mechanism underlying this induction is not known. Apoptotic processes in tumors could be responsible for clusterin gene activation. Alternatively, oncogenic mutations could modulate signal transduction, thereby inducing the gene. We examined the response of the rat clusterin gene to two oncogenes, Ha-ras and c-myc, in transfected Rat1 fibroblasts. While c-myc overexpression did not modify clusterin gene activity, the Ha-ras oncogene produced a seven to tenfold repression of clusterin mRNA; this down-regulation was also observed in the presence of c-myc. Since no induction of the clusterin gene was observed by the two oncogenes, we tested the alternative mechanism involving apoptosis. Growth factor withdrawal induced apoptosis, as shown by DNA degradation and micronuclei formation in the floating cells. Concomittantly we observed a three to tenfold increase in the amount of clusterin mRNA in the adhering cells of Rat1 and the c-myc transformed cell lines, and a weaker induction in the Ha-ras transformed cell line. On the basis of our results, we suggest that clusterin gene induction in the vital cells is produced by signaling molecules that are generated by the apoptotic cells. We conclude that apoptotic processes, not oncogenic mutations, are responsible for increased clusterin expression in tumors.

Animals↗
PubMed1997

Quartz crystal microbalances for quantitative biosensing and characterizing protein multilayers.

The use of quartz crystal microbalances (QCMs) for quantitative biosensing and characterization of protein multilayers is demonstrated in three case studies. Monolayers of QCM-based affinity biosensors were investigated first. Layers of a thiol-containing synthetic peptide constituting an epitope of the foot-and-mouse-disease virus were formed on gold electrodes via self-assembly. The binding of specific antibodies to epitope-modified gold electrodes was detected for different concentrations of antibody solutions. Oligolayers were studied in a second set of experiments. Dextran hydrogels were modified by thrombin inhibitors. The QCM response was used in a competitive binding assay to identify inhibitors for thrombin at different concentrations. Multilayers of proteins formed by self-assembly of a biotin-conjugate and streptavidin were investigated next. The QCM frequency response was monitored as a function of layer thickness up to 20 protein layers. A linear frequency decay was observed with increasing thickness. The decay per layer remained constant, thus indicating perfect mass coupling to the substrate. Frequency changes a factor of four higher were obtained in buffer solution as compared to measurements in dry air. This indicates a significant incorporation of water (75% weight) in the protein layers. This water behaves like a solid concerning the shear mode coupling to the substrate. The outlook discusses briefly the need for controlled molecular engineering of overlayers for subsequent QCM analysis, and the importance of an additional multiparameter analysis with other transducer principles and with additional techniques of interface analysis to characterize the mechanical coupling of overlayers as biosensor coatings. A promising trend concerns the use of QCM-arrays for screening experiments.

Biosensing Techniques↗
PubMed1996

A 'mixed' self-assembled monolayer for an impedimetric immunosensor.

A synthetic peptide with the amino acid sequence 135-154 of the capsid protein VP1 of the foot-and-mouth-disease virus was modified with omega-hydroxyundecanethiol and applied together with non-derivatised omega-hydroxyundecanethiol for consecutive adsorption onto gold electrodes according to self-assembling procedures. The binding of a specific antibody to prepared recognition layers could be monitored by measurement of impedance or capacitance. In order to avoid non-specific effects, all measurements were performed in the presence of BSA. The complex between the antigenic peptide and the antibody was split by applying 6 M urea solution. The gold electrodes were mounted into an optimised flow-through system in order to perform capacitance-time measurements. The immobilised peptide can be recognised repeatedly by specific antibodies.

Amino Acid Sequence↗