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R Bock

Publications and source records attributed to R Bock.

At least 55 records · Page 3Linked to original sources
PubMed1995

In vivo testing of a tobacco plastid DNA segment for guide RNA function in psbL editing.

A C-to-U RNA editing event creates a functional initiation codon for translation of the psbL mRNA in tobacco plastids. Small trans-acting guide RNAs (gRNAs) have been shown to be involved in editing site selection in kinetoplastid mitochondria. A computer search of the tobacco plastid genome (ptDNA) identified such a putative gRNA, a 14-nucleotide sequence motif that is complementary to the psbL mRNA, including the A nucleotide required to direct the C-to-U change. The critical A nucleotide of the putative gRNA gene was changed to G by plastid transformation. We report here that the introduced mutation did not abolish psbL editing. Since no other region of the plastid genome contains significant complementarity to the psbL editing site we suggest that, if gRNAs serve as trans-acting factors for plastid psbL mRNA editing, they either have only a limited complementarity to the editing site, or are encoded in the nuclear genome.

Base Sequence↗
PubMed1994

Introduction of a heterologous editing site into the tobacco plastid genome: the lack of RNA editing leads to a mutant phenotype.

The psbF mRNA is edited in spinach plastids by a C to U conversion, changing a serine to a conserved phenylalanine codon. In tobacco at this position a phenylalanine codon is present at the DNA level, and the psbF mRNA here is not edited. To test if the psbF editing capacity is evolutionarily conserved, the tobacco psbF gene was modified to match the corresponding spinach sequence. The endogenous tobacco gene was replaced with the modified copy using biolistic transformation. We report here that the heterologous editing site remains unmodified in transplastomic tobacco plants. The lack of editing is associated with slower growth, lowered chlorophyll content and high chlorophyll fluorescence, a phenotype characteristic of photosynthetic mutants. This finding confirms that the editing of the psbF mRNA is an essential processing step for protein function and thus provides direct proof for the biological significance of plant organellar RNA editing. Given that a mutant phenotype is associated with the lack of editing, it seems likely that the evolutionary loss of the site-specific capacity for psbF editing was preceded by the mutation that eliminated the editing requirement.

Amino Acid Sequence↗
PubMed1994

Isoelectric focusing and capillary zone electrophoretic studies using luteinizing hormone releasing hormone and its analog.

Iontophoresis is the movement of charged compounds into and through the skin under an external electromotive force. A molecule's charge and size, as well as other factors, will determine its ability to be iontophoresed. One can gain insight into certain electrical aspects of a molecule by determining its isoelectric point as well as its electrophoretic mobility. In the present study, isoelectric focusing and capillary zone electrophoretic studies of luteinizing hormone releasing hormone, (LHRH) and one of its analogs are performed in order to predict their ability to be iontophoresed. The pI of LHRH is determined to be 9.6 while that of its free acid is 6.9. In addition, it is concluded that although the two compounds are chemically similar, the native compound is more mobile at pH > 2.5. These results suggest that the native compound might be better suited for iontophoretic delivery than is the free-acid analog.

Buffers↗
PubMed1994

Pars tuberalis-specific cells in the ovine pituitary do express the common alpha-chain of glycoprotein hormones: an in situ hybridization and immunocytochemical study.

The ovine pituitary pars tuberalis was investigated by electron microscopy, immunocytochemistry and non-radioactive in situ hybridization in order to characterize further the pars tuberalis-specific cells, whose functional role within the endocrine system is still enigmatic. Ultrastructural analysis revealed that, besides gonadotropic cells, the vast majority of cells in the ovine pars tuberalis show the typical characteristics of pars tuberalis-specific cells with clear signs of secretory activity. Immunocytochemical staining with a polyclonal antibody directed against the alpha-subunit of ovine glycoprotein hormones and in situ hybridization with an antisense oligonucleotide complementary to the alpha-subunit mRNA showed that the common alpha-chain of glycoprotein hormones is expressed in pars tuberalis-specific cells. Antibodies against the beta-subunits failed to detect any of the known beta-chains of pituitary glycoprotein hormones in these cells. The demonstration of the glycoprotein alpha-subunit in pars tuberalis-specific cells of the adult sheep supports previously existing evidence that these cells do secrete one or more glycoprotein hormones. With respect to the thyrotropin-beta-like immunoreactivity in rats and hamsters, one might speculate that pars tuberalis-specific cells are a pluripotent cell type with a low secretory activity under basal conditions. Further studies should prove the hypothesis that a pre-existing mRNA pool in these cells can be used for sustained translation of glycoprotein hormones after physiological or pharmacological stimulation.

Animals↗
PubMed1993

Combined Ki-67 and Feulgen stain for morphometric determination of the Ki-67 labelling index.

In this note we present a combined Ki-67 and Feulgen stain for morphometric determination of the Ki-67 labelling index. The immunohistochemical part of this double staining technique is based on the alkaline-phosphatase-anti-alkaline-phosphatase (APAAP) method, visualizing the enzyme activity by the nitro-blue-tetrazolium chloride (NBT)/bromo-chloro-3-indolyl-phosphate (BCIP) technique. The NBT/BCIP complex resists the hydrolytic activity of the Feulgen stain. The staining method presented allows semi-automatic determination of both the total nucleus-area as well as the Ki-67 positive nucleus-area using a morphometric computer system. The Ki-67 labelling index thus achieved is based on the relative nuclear area of Ki-67 positive nuclei and is clearly more precise and efficient than the counting method using an ocular grid.

Alkaline Phosphatase↗
PubMed1993

Tissue- and stage-specific modulation of RNA editing of the psbF and psbL transcript from spinach plastids--a new regulatory mechanism?

The psbE operon of spinach chloroplasts, which includes the genes psbE, psbF, psbL and psbJ, encodes two RNA editing sites. One site corresponds to the initiation codon of the psbL transcript, as has been described earlier for the homologous transcript from tobacco, while at a second editing site, newly reported here, an internal phenylalanine codon of the psbF transcript is restored. Both these sites were investigated with respect to the extent of editing in spinach plastids at various developmental stages. The apparent existence of only completely edited transcripts in etioplasts and chloroplasts, indicates that light-induced processes are not acting as determinants in eliciting the editing process. Reduced editing is, however, observed in the psbF and psbL transcript from seeds and roots. This finding suggests that the RNA editing process is differentially down-regulated in leucoplasts and proplastids and that editing may, therefore, function as a regulatory device in plastid gene expression.

Amino Acid Sequence↗
PubMed1993

A new technique for post-embedding immunogold localization of vasopressin in the rat median eminence.

A modified cryosubstitution method is described which simultaneously preserves the ultrastructure and antigenicity of the osmium-sensitive neuropeptide vasopressin in rat median eminences. The organs were aldehyde-fixed, osmicated, and embedded in Epon. Thin sections were no longer needed to be etched and the titer of primary antiserum was near to the light microscopic level. Furthermore ice crystallization could not be observed.

Animals↗
PubMed1992

Renin-producing renal cell carcinomas--clinical and experimental investigations on a special form of renal hypertension.

The pathogenetic relationship between tumour and hypertension was investigated in 129 patients with renal cell carcinoma, of whom 41 (31.8%) were hypertensive. Of these 41 patients with renal tumours and hypertension, 6 (14.6%) were found to have primary reninism. In these patients the plasma renin activity in blood from the renal veins showed a tumour kidney to contralateral kidney ratio of between 4 and 7, and 2 patients also had secondary hyperaldosteronism. In the same 6 cases the renin content in the renal tumour tissue was significantly higher than that in tissue from the adjacent tumour-free renal cortex of the ipsilateral kidney. Immunohistochemical demonstration of renin in the tumour was only possible in these 6 cases. In 5 of these patients blood pressure returned to normal following nephrectomy; in the 6th case there was a drop in blood pressure after nephrectomy. In 3 renin-positive tumours examined, autonomous renin production was demonstrated in cell culture. Renin-producing renal cell carcinomas are an uncommon cause of renal hypertension. The differential diagnosis of hypertension should therefore also include renal tumour.

Captopril↗
PubMed1992

Polymyxin B: pharmacokinetics of single doses given intravenously and intramuscularly to turkeys, and minimal inhibitory concentrations for Escherichia coli and Pasteurella multocida.

The 50% and 90% minimal inhibitory concentrations (MIC50 and MIC90) of polymyxin B for avian Escherichia coli and Pasteurella multocida isolates were determined by the agar plate dilution method. Polymyxin B at approximate MIC level in serum was bactericidal for E. coli in 2 to 4 hours. Aqueous polymyxin B sulfate was administered by a single bolus intravenous injection into turkeys at 10,000 IU/kg, and by a single bolus intramuscular injection at 5,000, 10,000 or 20,000 IU/kg. Effective serum drug concentrations after intramuscular injection (MIC50 levels or greater) were maintained for E. coli for 7.0 hr (10,000 IU/kg) and 11.5 hr (20,000 IU/kg), and for P. multocida for 3.0 hr (10,000 IU/kg) and 4.1 hr (20,000 IU/kg). Pharmacokinetic parameters were calculated by non-compartmental methods. Elimination time half-lives, mean residence time, clearance, and apparent volume of distribution at steady state (Vdss) were all much higher for i.m. injection of 20,000 IU/kg than for i.m. injection of 10,000 IU/kg. We postulate that there exists a minimal tissue-interaction threshold concentration (MTC) at which polymyxin B can enter previously unavailable compartments or bind to previously refractory tissue components. Bioavailability of polymyxin B injected i.m. was 0.904 for the 10,000 IU/kg dose and 0.675 for the 20,000 IU/kg dose. Dosage intervals necessary to produce minimal steady state concentrations (Cssmin) equal to the MIC were calculated. Certain aspects of the use of the parameter Vdss, and limitations on the use of dosage interval calculations for polymyxin B, are discussed. One week after i.m. injection of polymyxin B at 10,000 IU/kg, high tissue drug levels were present, especially in bound form in liver. Following single injections, no toxic effects on turkeys were observed.

Animals↗
PubMed1992

Light microscopical demonstration of non-specific alkaline phosphatase activity with an incubation medium containing cerium and two calcium as the capturing agents. The cerium/calcium-hydrogen peroxide-P-phenylenediamine/pyrocatechol (Ce/Ca-H2O2-PPD/PC) double capture technique.

A new method for the light microscopical demonstration of alPase activity in cryotome sections by using simultaneously cerium and calcium as capturing agents (double capture technique) is described. This method has an increased sensitivity compared with the single cerium-based and the Gomori based-cerium (single calcium and cerium converted) with techniques described previously. Presuming that the enzymatic activity during incubation of sections in the presence of a defined capturing agent is constant, the increased sensitivity after employment of the double capture method could be attributed to a decrease of enzyme inhibition by cerium through the presence of calcium. Based on model experiments it is assumed that calcium phosphate and cerium phosphate are the primary reaction products, the former converting into cerium phosphate already during incubation. The remaining calcium phosphate is converted completely by treatment with cerium citrate solution (conversion reaction). After oxidation with H2O2 the cerium perhydroxyphosphate was visualized in a paraphenylenediamine/pyrocatechol (Hanker-Yates reagent) solution without H2O2 to give a black reaction product. This visualization procedure is superior to the DAB or DAB-Ni mode as published earlier. Some results concerning the mode of inhibition of the pseudoperoxidase activity of the hemoglobin are presented.

Alkaline Phosphatase↗
PubMed1992

[Carcinoma of the kidney with production of renin. A special form of hypertension].

A correlation between the tumour and hypertension was investigated in 129 patients with renal cancer. Forty-one patients (31.8%) suffered from hypertension. Primary increased renin secretion was detected in 6 of these patients (14.6%). The renin activity quotient measured in the renal veins between the renal tumour and the contralateral kidney was between 4 and 7. Secondary hyperaldosteronism was detected in only 2 of the patients with a significantly different renin activity in the renal veins. The renin level detected in the tumour itself of these 6 patients was significantly higher than the level detected in the parenchyma of the same kidney. Renin was demonstrated immunohistochemically in the tumours of these patients. The blood pressure returned to normal after nephrectomy in 5 patients and a marked fall in blood pressure was observed in the 6th patient. Cell cultures of 3 tumours revealed autonomous renin production. Renin-secreting renal carcinomas represent a rare form of renal hypertension, which is why the possibility of renal cancer should also be considered when investigating a case of hypertension.

Aldosterone↗
PubMed1991

Investigations of phagocytosis concerning the immunological defence mechanism of Mytilus edulis using a sublethal luminescent bacterial assay (Photobacterium phosphoreum).

1. A simple method for the determination of phagocytosis activity using mussel hemocytes by measuring the bioluminescence is presented. 2. The immunological defence activity based on phagocytosis is measured and quantified by a luminescent bacterial assay with Photobacterium phosphoreum. 3. The measuring system allows us to establish the stress of the immunological defence mechanism of organisms exposed to chemicals and polluted rivers or sewage. Results with reference substances and the phagocytosis indices of exposed mussels from Norwegian aquaculture plants compared to those of mussels from the German Wadden Sea are given as examples.

Animals↗