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Biomedical subjects

H J Keller

Publications and source records attributed to H J Keller.

13 recordsLinked to original sources

Automated and computer-controlled method for the measurement of the crystallization of calcium oxalate monohydrate in urine.

Monitoring of crystallization of calcium salts with ion-selective electrodes has turned out to be a very sensitive method. The difficulties of handling these electrodes in native whole urine and other biological fluids have been eliminated by new calcium analyzers, which clean and calibrate the electrodes after each measurement. To study crystallization kinetics, repeated calcium ion measurements have to be performed at regular intervals. For this purpose we have developed a special sampler and software. The sampler brings a thermostat-controlled crystallization chamber to the analyzer at preselected intervals. The computer directs and coordinates the sampler and the analyzer, stores the received results and prints out growth curves. Furthermore it calculates the half-time (h) and the maximum decrease of ionic calcium at infinite incubation time (delta Ca2+ infinity). Both values are shown to characterize the growth of calcium oxalate monohydrate in urine. Results are obtained within 40 min.

Calcium↗

Interaction of Xenopus TFIIIC with the TFIIIA.5 S RNA gene complex.

The general transcription factor TFIIIC is necessary for transcription initiation by RNA polymerase III. TFIIIC binds predominantly to the B-Block promoter element, which is present in tRNA genes, several viral RNA genes and repetitive DNA elements, and to the TFIIIA.DNA complex on 5 S RNA genes. Here we report a characterization of Xenopus laevis TFIIIC and its interaction with the TFIIIA.5 S RNA gene complex. A polypeptide with apparent molecular mass of 85 kDa was specifically cross-linked to a B-Block oligonucleotide by UV light. This polypeptide was present in the partially purified TFIIIC fraction and in a complex with a B-Block double-stranded oligonucleotide isolated by nondenaturing gel electrophoresis. TFIIIC.TFIIIA.DNA gel mobility shift complexes were obtained using B-Block DNA affinity-purified TFIIIC and buffer conditions employing low Mg2+ (1 mM) and high dithiothreitol (7 mM) concentrations. Three TFIIIC.TFIIIA.5 S RNA gene complexes were observed by gel mobility shift analysis. One of these complexes was resistant to dissociation by the addition of competing DNA, but the formation of all three complexes was prevented by the inclusion of excess specific competitor DNA in the initial binding reactions. The apparent affinity of TFIIIC for the TFIIIA.5 S DNA complex was 5-fold higher for the somatic-type 5 S RNA gene than for the oocyte-type 5 S RNA gene. Mutations near the 5' boundary of the TFIIIA binding site alter the DNase I footprint of the TFIIIA.DNA complex and reduce the affinity of TFIIIA-mutant 5 S gene complexes for TFIIIC. Differences in TFIIIC affinity for the two classes of 5 S RNA genes may play a role in the developmental regulation of these gene families.

Animals↗

Additional intragenic promoter elements of the Xenopus 5S RNA genes upstream from the TFIIIA-binding site.

The major promoter element of the Xenopus laevis 5S RNA gene is located within the transcribed region of the gene and forms the binding site for the transcription initiation factor TFIIIA. We report an analysis of deletion and substitution mutations within the coding region of the major oocyte-type 5S gene of X. laevis. Our results differ from those of previous mutagenesis studies conducted on the somatic-type genes of Xenopus borealis and X. laevis. Transcription assays in whole oocyte S-150 extracts, with both oocyte- and somatic-type mutants, revealed additional promoter elements between the start site for transcription and the binding site for TFIIIA. These sequences regulate the efficiency of binding TFIIIC, a transcription factor required by the genes transcribed by RNA polymerase III containing intragenic promoters. Under TFIIIC-limiting conditions, the somatic-type gene had a 10-fold-higher affinity for TFIIIC than did the major oocyte-type 5S gene. One mutation in the oocyte-type gene (nucleotides +33 to +39) reduced TFIIIC affinity and transcriptional activity four- to fivefold. Differences in TFIIIC affinity between oocyte- and somatic-type genes may contribute to the differential transcription of these genes observed during Xenopus embryogenesis.

Animals↗

[Incidence of hemorrhage following suprapubic Cystofix puncture using a new trocar-cannula combination--a randomized, prospective comparative study].

In this study a new trocar-cannula-unit(10 French) is introduced, which was used in a prospective randomised comparative study in 74 patients for suprapubic bladder punction. Another 75 patients served as controls receiving a suprapubic punction with the standard Cystofix (French 10)-system. In the control-group 9 cases (12%) of macrohematuria occurred. On the other hand we found that the incidence of macrohematuria was reduced to 5 cases (6.8%), when using the new trocar-cannula-unit, which had been developed in our hospitals.

Adult↗

Cysteine: depolarization-induced release from rat brain in vitro.

Compounds released on depolarization in a Ca2+-dependent manner from rat brain slices were screened to identify candidates for neuroactive substances. Lyophilized superfusates were analyzed by reversed-phase HPLC after derivatization with 9-fluorenyl N-succinimidyl carbonate. One of the compounds that showed an increase of concentration in superfusates in the presence of iodoacetamide was identified as the cysteine (Cys) derivative, S-carboxamidomethylcysteine, by fast atom bombardment mass spectrometry and other methods. This stable Cys derivative originates from endogenous, extracellular Cys. The finding led to a method for quantification of Cys in superfusates by immediate cooling of the superfusates to 0 degrees C and reaction of Cys with N-ethylmaleimide. Depolarization-induced Ca2+-dependent release of Cys was most prominent in the neocortex, followed by the mesodiencephalon, striatum, and cerebellum. This suggests that Cys is released from a neuronal compartment and might be involved in neurotransmission.

Animals↗

Antitumor activity of cis-dihalogenobis(1-phenyl-1,3-butanedionato) titanium(IV) compounds against Walker 256 carcinosarcoma. A new class of antineoplastic agents.

The antitumor activity of cis-dihalogenobis(1-phenyl-1,3-butanedionato)titanium(IV) compounds against Walker 256 carcinosarcoma in comparison to cis-dichlorodiammineplatinum(II) (cisplatin, cis-DDP, NSC 119875) was investigated. The tested compounds showed in this experimental model an enhanced median survival time and a lower toxicity compared to cisplatin.

Animals↗