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Biomedical subjects

P Kaspar

Publications and source records attributed to P Kaspar.

At least 19 recordsLinked to original sources
PubMed2001

Positive and negative regulation of myogenic differentiation of C2C12 cells by isoforms of the multiple homeodomain zinc finger transcription factor ATBF1.

The ATBF1 gene encodes two protein isoforms, the 404-kDa ATBF1-A, possessing four homeodomains and 23 zinc fingers, and the 306-kDa ATBF1-B, lacking a 920-amino acid N-terminal region of ATBF1-A which contains 5 zinc fingers. In vitro, ATBF1-A was expressed in proliferating C2C12 myoblasts, but its expression levels decreased upon induction of myogenic differentiation in low serum medium. Forced expression of ATBF1-A in C2C12 cells resulted in repression of MyoD and myogenin expression and elevation of Id3 and cyclin D1 expression, leading to inhibition of myogenic differentiation in low serum. In contrast, transfection of C2C12 cells with the ATBF1-B isoform led to an acceleration of myogenic differentiation, as indicated by an earlier onset of myosin heavy chain expression and formation of a higher percentage of multinucleated myotubes. The fourth homeodomain of ATBF1-A bound to an AT-rich element adjacent to the E1 E-box of the muscle regulatory factor 4 promoter mediating transcriptional repression. The ATBF1-A-specific N-terminal region possesses general transcription repressor activity. These results suggest that ATBF1-A plays a role in the maintenance of the undifferentiated myoblast state, and its down-regulation is a prerequisite to initiate terminal differentiation of C2C12 cells.

Animals↗
PubMed1999

Myb-interacting protein, ATBF1, represses transcriptional activity of Myb oncoprotein.

Using the yeast two-hybrid system, the transcription factor ATBF1 was identified as v-Myb- and c-Myb-binding protein. Deletion mutagenesis revealed amino acids 2484-2520 in human ATBF1 and 279-300 in v-Myb as regions required for in vitro binding of both proteins. Further experiments identified leucines Leu325 and Leu332 of the Myb leucine zipper motif as additional amino acid residues important for efficient ATBF1-Myb interaction in vitro. In co-transfection experiments, the full-length ATBF1 was found to form in vivo complexes with v-Myb and inhibit v-Myb transcriptional activity. Both ATBF1 2484-2520 and Myb 279-300 regions were required for the inhibitory effect. Finally, the chicken ATBF1 was identified, showing high degree of amino acid sequence homology with human and murine proteins. Our data reveal Myb proteins as the first ATBF1 partners detected so far and identify amino acids 279-300 in v-Myb as a novel protein-protein interaction interface through which Myb transcriptional activity can be regulated.

Amino Acid Sequence↗
PubMed1993

Identification of CpG island at the 5' end of murine leukemia inhibitory factor gene.

We identified a CpG island at the 5' end of murine leukemia inhibitory factor gene (LIF). The CpG island is 0.6 kb long and covers most of the first exon and first intron. The region is non-methylated, its G+C content is 65% and relative frequency of CpG dinucleotide is 0.7. The block of 150 nucleotides, which is 72% conserved between murine, human, ovine and porcine genes, is a part of the CpG island. Two DNA fragments from this CpG island interact with nuclear proteins from NIH 3T3 cells. One fragment partially covers the block of conserved nucleotides. Human, ovine and porcine LIF genes also contain G+C- and CpG-rich sequences in the corresponding region.

Animals↗
PubMed1992

Blastocyst implantation depends on maternal expression of leukaemia inhibitory factor.

A critical point during mammalian pregnancy is the implantation of the blastocyst when the embryo attaches to the wall of the uterus. The autonomously developing preimplantation embryo then becomes dependent on the maternal environment for its continued development. Little is known about the regulation of implantation, except that a complex interaction between peptide and steroid hormones synchronizes the preparation of the uterus for implantation with the development of the embryo. Whether the implantation event is under maternal or embryonic control is also unclear (reviewed in refs 1, 2). We have previously shown that a cytokine, leukaemia inhibitory factor (LIF), is expressed in the uterine endometrial glands specifically on the fourth day of pregnancy. This burst of expression is under maternal control and always precedes implantation of the blastocyst. Here we report that transient expression of LIF in mice is essential for implantation. Females lacking a functional LIF gene are fertile, but their blastocysts fail to implant and do not develop. The blastocysts, however, are viable and, when transferred to wild-type pseudopregnant recipients, they can implant and develop to term.

Animals↗
PubMed1992

Results of the multicenter evaluation of the CEDIA Theophylline assay.

We report on the results of the multicenter evaluation of the CEDIA Theophylline assay on Boehringer Mannheim/Hitachi analyzers in 15 clinical laboratories in Europe and U.S.A. Main items of investigation were imprecision, recovery of control sera, interlaboratory survey and method comparisons using patient samples. Imprecision was found to be comparable to other routine methods. An advantage of the CEDIA assay can be seen in the good interlaboratory transferability of results. The new test has been shown to measure very accurately particularly by comparison with HPLC procedures revealing highly correspondent results. The reagent can be used up to one month using multiple recalibration. Due to its high practicability and reliability the CEDIA Theophylline assay can be recommended as a very suitable routine method for therapeutic drug monitoring on random access analyzers like Boehringer Mannheim/Hitachi analysis systems.

Dose-Response Relationship, Drug↗
PubMed1992

CEDIA--homogeneous immunoassays for the 1990s and beyond.

New homogeneous enzyme immunoassays have been developed for cortisol, digoxin, digitoxin, theophylline, phenytoin, and phenobarbital using the cloned enzyme donor immunoassay technology. As applied to Boehringer Mannheim/Hitachi analysis systems these methods provide rapid, accurate and precise quantification of analytes, with minimal interferences from endogenous serum constituents and low cross-reactivities to structurally-related hormonal precursors, drug metabolites and natural compounds. Additional significant features of the new assays are linear standard curves and two-point calibration. The six CEDIA assays join the two currently available CEDIA assays for determination of the thyroid parameters T4 and T Uptake. Additional new therapeutic drug and anemia monitoring assays are under development, demonstrating the versatility of the cloned enzyme donor immunoassay technology. These tests, in concert with Boehringer Mannheim/Hitachi analyzers, provide a high throughput, random access immunoassay system. The menu of available assays should continue to increase during the 1990s, providing efficient automation while allowing consolidation of testing on a limited number of instrument systems.

Digitoxin↗
PubMed1992

Results of the multicenter evaluation of a homogeneous immunoassay for digitoxin based on the cloned enzyme donor immunoassay technology.

We evaluated a CEDIA assay for the determination of digitoxin in serum on random access analyzers. The multicenter evaluation included studies on the analytical range, calibration stability and reproducibility of the new assay. Moreover, recovery in controls, transferability of results obtained in different laboratories, comparability with routine methods, and the effect of various interfering factors have been analyzed. Summarized the analytical performance was comparable to that of routine methods. The CEDIA Digitoxin assay represents an attractive alternative to established digitoxin immunoassays because it can be performed on random access analyzers, thus permitting the simultaneous determination of digitoxin and other serum analytes without sample splitting.

Digitoxin↗
PubMed1992

Evaluation of a simple screening test for the quality of drinking water systems.

A simple and inexpensive assay for detection of H2S producing bacteria in drinking water was evaluated in Paraguay. Using an improved preparation procedure, results were basically temperature independent in the range of 22-37 degrees C and correlated well with the presence of total coliform bacteria (96%, 28 degrees C). The assay was not suitable for control of surface water and dug well water due to the frequent presence of non-faecal coliforms. However, it was very suitable for routine control of high quality water systems, like treated community water systems or deep-tube well water, where complete absence of coliforms is required. The H2S-test is very inexpensive and does not require a laboratory technician or special equipment. Its use permits a cost-effective routine control of high quality community water systems in developing countries. In addition, this test turned out to be an effective educational tool for heightening the awareness of people towards dangers of unclean drinking water.

Bacteriological Techniques↗
PubMed1991

[Carotenemia].

The authors demonstrate two infants with a yellow skin coloration which developed as a result of excessive dietary carotenoid intake. The yellow coloration, contrary to jaundice, did not affect the sclerae. Elevated blood carotenoid levels were found and a temporary increase of aminotransferases. In both infants lower IgA levels were recorded. After modification of the diet the yellow coloration disappeared and gradually the aminotransferases reached normal levels.

Carotenoids↗
PubMed1988

Expression and properties of prochymosin derivatives containing extensions of various length in the pro-part.

A series of hybrid prochymosin derivatives containing portions of the simian virus 40 small-t antigen in the pro-part was constructed. Portions comprising 93, 63, 47, 12, and 1 amino acid (aa) from the N terminus of the small-t antigen were separately fused via eight polylinker-encoded amino acids to a prochymosin product commencing with the 5th aa of the pro-part. All the DNAs coding for the hybrid proteins were put under pL-promoter control in the expression constructs. Expression revealed that only fusion of the 47-aa or 12-aa stretch of the small-t antigen to prochymosin gave stable protein products and that only the latter one allowed the hybrid prochymosin to be activated to chymosin. The products containing 93 aa and 63 aa of small-t antigen were unstable and degraded. Complete removal of the small-t antigen portion led to mRNA instability, probably owing to inefficient initiation of translation.

Base Sequence↗
PubMed1988

Application of sandwich immunoassays for the determination of sample-specific background signals and its use for calibration of immunoassays.

Sample-related background signals in immunoassays can be measured by a variation of the double antibody sandwich principle, in which the unlabelled specific antibody is substituted by a similar unrelated non-specific antibody. This permits differentiation between the analyte-specific and the background signal components for each sample. The method permits selection of sera with no or low analyte content for use as analyte diluent and for defining the zero point of the calibration curve. The method also permits control of analyte content during production processes which may change the background signal as well as identification of samples with atypical background signals. The procedure has been used for the calibration of enzyme immunoassays for alpha-fetoprotein (AFP) and human thyroid-stimulating hormone (TSH).

Animals↗
PubMed1988

[Inhalative immunostimulation in patients with chronic bronchitis].

80 patients with chronic bronchitis were treated inhalatively for a time of 3 weeks with a polyvalent bacterial lysate (IRS 19) or with placebo. The study was carried out in a double-blind fashion. Treatment with IRS 19 resulted in improved bronchoscopic findings compared with the pretherapy findings. Patients with bronchial hyperreactivity showed in the IRS 19 group small improvement of the hyperreactivity. In this group the contents of IgG in the BAL fluid showed a significant reduction, the other parameters were unchanged.

Adjuvants, Immunologic↗
PubMed1987

Subcloning of prochymosin cDNA for Plac controlled expression.

Two overlapping segments of prochymosin cDNA clones (Liebscher et al., 1985) were used to construct plasmids that expressed an activable zymogen product and thus verified the integrity of the reverse transcripts. The pUC9 vector was used for the expression, under the control of the lac promoter. The expression product (a fused protein consisting of the N-terminus of beta-galactosidase, a polylinker-coded peptide and prochymosin from its 5th amino acid) displayed, upon activation by the usual procedure, the properties of calf chymosin. The active product was identified by milk-clotting tests, "caseinography" and protein electrophoresis of immunoprecipitates. The "boxing" of prochymosin cDNA in the constructed plasmids makes them a versatile source of this cDNA for other expression constructs.

Chymosin↗