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Biomedical subjects

K Thiele

Publications and source records attributed to K Thiele.

At least 19 recordsLinked to original sources

Induction of aminopeptidase N/CD13 on human lymphocytes after adhesion to fibroblast-like synoviocytes, endothelial cells, epithelial cells, and monocytes/macrophages.

Aminopeptidase N (APN/CD13) is a transmembrane ectoenzyme occurring on a wide variety of cells. In contrast to monocytes and granulocytes, lymphocytes of peripheral blood do not express CD13 Ag. However, tumor-infiltrating T cells in renal cell cancer as well as synovial fluid T cells from patients suffering from various forms of arthritis can be CD13 positive. To learn more about expression of CD13 in these tissues, we cocultured lymphocytes with different adherent cell lines. CD13 expression was induced in T and B lymphocytes upon adhesion to fibroblast-like synoviocytes, HUVEC, renal tubular epithelial cells, and monocytes/macrophages but not always upon interaction with different tumor cell lines. Induction of APN was rapid, occurring as early as 1 h after coincubation. Expression persisted for >3 days and partially resisted inhibition by cycloheximide. Fixation of adherent cells with paraformaldehyde could not prevent induction of CD13 in lymphocytes. Soluble APN from human kidneys or placenta could not induce CD13 expression on lymphocytes. Induction of CD13 Ag on lymphocytes required direct cell-to-cell contact as shown in experiments using dual chambers. Lymphocytes exhibited an induction not only in CD13 protein but also in Ala-pNA-cleaving enzyme activity and in CD13 mRNA. Lymphocytic expression of CD13 represents a potentially increased cellular ability to inactivate inflammatory mediators. Furthermore, CD13 could be involved in adhesion, in lymphocytic migration, or in the Ag processing of peptides bound in the groove of MHC class II molecules.

B-Lymphocytes

Treatment of fibroblast-like synoviocytes with IFN-gamma results in the down-regulation of autotaxin mRNA.

In an effort to isolate genes that change expression at the mRNA level during treatment of fibroblast-like synoviocytes (SFC) with IFN-gama, we performed a differential display analysis. Here, we report the isolation of a cDNA clone corresponding to a 3.1 kb mRNA species that is reduced in synoviocytes after culture with IFN-gama. Sequence analysis revealed the 211 bp length cDNA clone to be identical to the motility-stimulating 125 kDa protein autotaxin (ATX). The down-regulation of ATX mRNA was confirmed by Northern blot analysis as well as competitive RT-PCR. SFC express 1 ng ATX mRNA/microgram total RNA. IFN-gama down-regulated ATX mRNA up to 50% as compared to control. Our results add a new finding to the manifold functions described for IFN-gama in rheumatoid arthritis.

Base Sequence

The promoter of the H1zero histone gene contains a DNA element bound by retinoic acid receptors.

Retinoic-acid mediated differentiation of F9 cells is accompanied by an increased transcription of the histone H1zero gene. This increase is an early response after addition of retinoic acid, suggesting a direct effect of the hormone on transcription of the gene. We show now that the promoter of histone H1zero contains a DNA element, localized 531 base-pairs upstream of the cap site, that is composed of a direct repeat of the sequence PuGGTCA separated by eight base-pairs. This element confers retinoic acid responsiveness to a heterologous thymidine kinase promoter in F9 and HeLa cells. Furthermore, the element forms retarded complexes not only with bacterially expressed retinoic acid receptors (RARs) and retinoid X receptors (RXRs), but also with endogenous F9 receptors. Our results suggest therefore that retinoic acid receptors can control the expression of a chromatin structural gene the expression of which is associated with a differentiated phenotype.

Animals

Persistent replication of herpes simplex virus type 1 in JOK-1 cells.

Infection of the human B cell line JOK-1 with herpes simplex virus type 1 persisted over a period of more than 12 months (to date). Although limited cytopathic effects were seen, viral infection did not lead to extinction of the culture. Infectious centre assays, performed at various times after infection, revealed that only a small proportion of cells (1 to 10%) produced infectious virus particles. However, immunofluorescence studies showed that at any given time considerably more cells than calculated by infectious centre assays contained the immediate early viral protein ICP4 and expressed viral glycoproteins. These observations were confirmed by in situ hybridization analyses which revealed the presence of viral DNA even in cells not producing infectious particles. Since no evidence for the involvement of interferon could be found, some other so far unknown intrinsic property of the cells must be responsible for the restriction of virus replication and/or maturation.

Antigens, Viral

Effects of different interferons on the replication of herpes simplex virus in human T lymphocytes.

Human T cells were activated with PHA and after 48 h they were treated with preparations of different human interferons (IFNs). After a further incubation period of 24 h, the cells were washed and infected with herpes simplex virus type 1 (HSV-1). Reduced virus titers were observed in T cells pretreated with IFN-alpha or IFN-beta, whereas IFN-gamma showed no antiviral effects. These findings indicate that IFN-gamma does not exert protection against viral infection in this system. We investigated the synthesis of HSV-coded early and late proteins in T cells pretreated with IFN-alpha and IFN-beta. Immunofluorescence studies revealed inhibition of expression of the immediate early alpha-protein ICP 4. Induction of DNA-polymerase, a viral beta-protein, was inhibited both by IFN-alpha and IFN-beta in a dose-dependent manner. As suggested by SDS-polyacrylamide gel electrophoresis, other viral beta- and gamma-proteins of HSV were inhibited by IFN-alpha and IFN-beta as well.

Cell Division

The role of prostacyclin in the hypercapnic and hypoxic cerebrovascular dilations.

The cerebral blood flow (CBF H/A) and the production of a stable prostacyclin metabolite, 6-Keto PGF 1 alpha ( 6KPGF ) was studied in 5 baboons in control, hypercapnic and hypoxic conditions. In steady-state conditions CBF H/A was measured by the clearance of an intra-arterial bolus injection of 133xenon and arterial and cerebral venous blood was sampled for assay of 6KPGF by radioimmunoassay. Both hypercapnia and hypoxia significantly increased CBF H/A and both increments were abolished by indomethacin. However, only hypoxia showed an increased 6KPGF production. Thus, hypoxia, but not hypercapnia, appears to produce cerebral vasodilation by increasing prostacyclin production.

6-Ketoprostaglandin F1 alpha

[New biologically active theophylline derivatives. Synthesis and pharmacologic properties of flufylline and fluprofylline].

Within the course of the research project for finding new cardiovascular drugs we have synthetized since 1976 piperazine- and piperidine derivatives of theophylline. The screening shows for the substances Sgd 19578 (flufylline) and Sgd 14480 (fluprofylline) a long-lasting blood pressure lowering activity as well as a remarkable serotonin- and histamine antagonism and broncholytic activity. On the basis of these results these compounds were selected for further investigation.

5-Hydroxytryptophan

Sgd 101/75: a sympathomimetic that can be used to identify a new subtype of alpha-adrenoceptor, the alpha 1s-adrenoceptor.

When Sgd 101/75 was compared with clonidine in a number of tests for CNS activity, Sgd 101/75 exhibited little activity in any test. Sgd 101/75 raised BP without affecting HR in several species of anaesthetised animals. The rise in BP was subject to tachyphylaxis, could be antagonised by alpha 1-adrenoceptor antagonists, and was obtainable in reserpinised animals. The vasopressor effect of NA was antagonised by Sgd 101/75. Thus Sgd 101/75 is a directly acting partial agonist for vascular alpha 1-adrenoceptors. On the coaxially stimulated guinea-pig ileum and field stimulated rat vas deferens, the twitch response to single pulse stimulation was reduced by NA or clonidine stimulating prejunctional alpha 2-adrenoceptors. Sgd 101/75 antagonised these inhibitory effects competitively (pA2 for antagonism of clonidine on the vas = 6.12). Sgd 101/75 acted as a specific partial agonist on the alpha 1-adrenoceptors of the guinea-pig taenia caecum that subserve relaxation of this tissue. Sgd 101/75 was a full agonist on the alpha 1-adrenoceptors of the rat anococcygeus in vitro. Phenoxybenzamine (300 pM for 30 min, followed by 20 washes over the next 30 min) reduced contractions of the anococcygeus to Sgd 101/75, but produced little inhibition of NA-induced contractions. In phenoxybenzamine-pretreated preparations, Sgd 101/75 (400 microM) did not antagonise NA (maximal effect and EC50 values not changed significantly), so it was concluded that Sgd 101/75 and NA interact with different alpha 1-adrenoceptor subtypes in this tissue. The subtype specifically activated by Sgd 101/75 was designated the alpha 1s-adrenoceptor. The mouse anococcygeus contained alpha 1s-adrenoceptors, whereas this receptor was absent from the rabbit anococcygeus.

Adrenergic alpha-Agonists

[New cerebrally active basic dithienyl compounds (author's tranls)].

A review on new cerebrally active basic dithienyl compounds related to (+)-(R)-alpha-((S)-1-[(3,3-di-3-thienylallyl)amino]-ethyl)-benzylalcohol (tinofedrine) is presented. Tinofedrine was selected out of a large number of related compounds on account of its high increase of cerebral blood flow, its improvement of heart performance, of the metabolism of the brain, and because it is well toleraded. Different routes of synthesis are discussed.

Animals

[Spectroscopic characterisation of tinofedrine (author's transl)].

With the aid of spectroscopical methods the structure of (+)-(R)-alpha-[(S)-1-[(3,3-di-3-thienylallyl)amino]-ethyl]-benzyl-alcohol (tinofedrine, 1), a new cerebrally active compound, was confirmed. The relative and absolute configuration is discussed in view of spectroscopic data.

Chemical Phenomena

[Beclobrate and eniclobrate hydrochloride, new diphenylmethane derivatives as agents for lowering cholesterol and triglyceride levels. Part I: Synthesis and consideration of structure-activity relationships (author's transl)].

Within the course of a research project for finding new lipid-lowering substances with better therapeutic indices than the standard agent in use, various diphenylmethane derivatives were synthesized and tested with respect to their activity and toxicity. On the basis of these results Sgd 24774 (beclobrate) and Sgd 33374 (eniclobrate-hydrochloride) were selected for further investigation and clinical studies.

Animals

[A contribution to the stereospecific synthesis of antifungal imidazolyloxime-ethers/Oxiconazole nitrate (Sgd 301-76), a new broadspectrum antifungal agent (author's transl)].

A stereospecific synthesis of antifungal imidazolyloxime-ether derivatives is reported. The compound Sgd 301-76 (oxiconazole nitrate) = (Z)-1-(2,4-dichlorophenyl)-2-(1H-imidazol-1-yl)-O-(2,4-dichlorobenzyl)-ethanoneoxime-nitrate was selected for clinical trials. The (E)- and (Z)-assignments, respectively, were made on the basis of 1-H-NMR-spectral data.

Antifungal Agents