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T Müller

Publications and source records attributed to T Müller.

At least 361 records · Page 20Linked to original sources
PubMed1994

Isolation, biochemical characterization and N-terminal sequence of rolipram-sensitive cAMP phosphodiesterase from human mononuclear leukocytes.

A cyclic AMP specific phosphodiesterase (type IV) was purified 450,000-fold from human peripheral blood mononuclear cells through a sequence of chromatographic steps involving anion exchange, affinity chromatography on a matrix coupled to a derivative of the type IV inhibitor rolipram, and gel filtration. The enzyme showed apparent molecular masses of 70 kDa on gel filtration and 35 kDa on denaturing or native PAGE, indicating a possible dimerization or cleavage under certain conditions. The isoelectric point was 4.6. Kinetic parameters were Km = 2.2 microM, Ki = 1.2 microM (rolipram) and vmax = 80 mumol/min per mg protein. The most probable N-terminal sequence was determined as SLTNTNIPRF, 80% identical to part of the deduced amino acid sequence from cDNA sequences of PDE IVA and PDE IVD.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1994

Cloning and characterization of the mouse gene encoding mammary-derived growth inhibitor/heart-fatty acid-binding protein.

From a mouse genomic library we isolated and characterized a gene, Fabph1, encoding mammary-derived growth inhibitor (MDGI)/heart fatty-acid-binding protein (H-FABP). Exon sequences were identical with a MDGI-encoding cDNA isolated previously from the mammary gland of pregnant mice. The product of this gene has also been detected in heart, where it had been termed H-FABP. It has an intron/exon structure similar to other FABP-encoding genes. In addition to this expressed gene, we isolated a related intronless pseudogene, Fabph-ps, with an open reading frame which was highly conserved when compared with Fabph1. Fabph1 was positioned on chromosome (Chr) 4 using interrelated sequence locus, Fabph-rs1, to Chr 8. A Mus spretus-specific related sequence, Fabph-rs2, was identified on Chr 17 by analysis of interspecies crosses. The 5'-flanking region of Fabph1 contains putative transcription factor-binding elements which could account for its constitutive expression in muscle tissue, as well as for its developmental stage-dependent expression in mammary epithelium.

Amino Acid Sequence↗
PubMed1994

Aspects of receptor binding and signalling of interleukin-4 investigated by site-directed mutagenesis and NMR spectroscopy.

Cytokines are hormones that carry information from cell to cell. This information is read from their surface upon binding to transmembrane receptors and by the subsequent initiation of receptor oligomerization. An influence on this process through mutagenesis on the hormone surface is highly desirable for medical reasons. However, an understanding of hormone-receptor interactions requires insight into the structural changes introduced by the mutations. In this line structural studies on human IL-4 and the medically important IL-4 antagonists Y124D and Y124G are presented. The site around Y124 is an important epitope responsible for the ability of IL-4 to cause a signal in the target cells. It is shown that the local main-chain structure around residue 124 in the variants remains unchanged. A strategy is presented here which allows the study of these types of proteins and their variants by NMR which does not require carbon labelled samples.

Amino Acid Sequence↗
PubMed1994

Radio-frequency microtools for particle and liver cell manipulation.

Single particles can be manipulated by applying high frequencies to ultramicro electrode arrays fabricated on planar structures. Heat production can be reduced to the extent that intense electric fields can be applied even to unmodified cell culture media. Animal cells grow normally in the high field (up to 100 kV/m) between such continuously energized multielectrodes. As with laser tweezers [1-3], this technique can capture particles and cells in field traps, generate linear movement, and permit cell cultivation. It can also produce micropatterns of pH gradients, field-cast objects, and control cell adhesion. These microtools may be combined to develop cell separators, microsensors, and controlled-biocompatibility surfaces.

Animals↗
PubMed1994

A new perineal template assembly for high-dose-rate interstitial brachytherapy of gynecologic malignancies.

This technical note introduces a modified Syed-Neblett type perineal template for high-dose-rate interstitial brachytherapy of gynecologic malignancies. The template can easily be disassembled after the insertion of the central needles into the pelvis allowing the cystoscopic and rectoscopic control of the needle positions. Thus needles penetrating the bladder or the rectum are recognized and can be repositioned before reassembling the template. Elimination of hot irradiation zones in tumor-free bladder and rectum walls might lead to reduction of severe late complications without compromising the therapeutic efficacy.

Adult↗
PubMed1994

Members of the fatty acid binding protein family are differentiation factors for the mammary gland.

Mammary gland development is controlled by systemic hormones and by growth factors that might complement or mediate hormonal action. Peptides that locally signal growth cessation and stimulate differentiation of the developing epithelium have not been described. Here, we report that recombinant and wild-type forms of mammary-derived growth inhibitor (MDGI) and heart-fatty acid binding protein (FABP), which belong to the FABP family, specifically inhibit growth of normal mouse mammary epithelial cells (MEC), while growth of stromal cells is not suppressed. In mammary gland organ culture, inhibition of ductal growth is associated with the appearance of bulbous alveolar end buds and formation of fully developed lobuloalveolar structures. In parallel, MDGI stimulates its own expression and promotes milk protein synthesis. Selective inhibition of endogenous MDGI expression in MEC by antisense phosphorothioate oligonucleotides suppresses appearance of alveolar end buds and lowers the beta-casein level in organ cultures. Furthermore, MDGI suppresses the mitogenic effects of epidermal growth factor, and epidermal growth factor antagonizes the activities of MDGI. Finally, the regulatory properties of MDGI can be fully mimicked by an 11-amino acid sequence, represented in the COOH terminus of MDGI and a subfamily of structurally related FABPs. This peptide does not bind fatty acids. To our knowledge, this is the first report about a growth inhibitor promoting mammary gland differentiation.

Amino Acid Sequence↗
PubMed1994

Heme oxygenase expression in Swiss 3T3 cells following exposure to aqueous cigarette smoke fractions.

A dose-dependent and transiently elevated expression of a cytoplasmic 32 kDa protein was observed in Swiss albino 3T3 fibroblasts exposed to mainstream cigarette smoke (CS) trapped in phosphate-buffered saline solutions (smoke-bubbled PBS). The protein was identified as heme oxygenase (HO) (heme, hydrogen donor:oxygen oxidoreductase, EC 1.14.99.3) by Western blotting using an anti-rodent HO-specific antibody. Kinetic investigations revealed that HO protein and its mRNA were detectable in smoke-bubbled PBS-treated cells between 1 and 24 h after exposure to 0.03 puffs (approximately 1 cm3) CS per ml medium. As a result of transcriptional activation, a nearly 50-fold increase in the amount of HO mRNA was determined after 8 h exposure compared to control levels. Since literature data indicate that there is a link between glutathione depletion and HO expression, the same was assumed for cells exposed to smoke-bubbled PBS, as a decrease of more than 60% in glutathione levels was observed after the exposure. This was further supported by the observation that no elevated amounts of HO mRNA appeared in smoke-bubbled PBS-treated cells when cysteine was exogenously added. However, although these effects may be attributable to the formation of hydroxyl radicals (which have been shown to induce HO and to deplete glutathione levels and which appear in aqueous smoke-containing solutions via the iron-catalysed Fenton reaction) neither catalase nor the iron cation chelating agent o-phenanthroline were able to suppress or even to reduce HO expression in smoke-bubbled PBS-treated cells. On the contrary, at comparable concentrations both compounds were found to be potent inhibitors of smoke-dependent DNA strand breaks. Hence, reactive species other than Fenton reaction-derived hydroxyl radicals are responsible for the effects observed in the present study.

3T3 Cells↗
PubMed1994

The expression pattern of a novel gene encoding brain-fatty acid binding protein correlates with neuronal and glial cell development.

Fatty acid binding proteins (FABPs) are a multigene family of small intracellular proteins that bind hydrophobic ligands. In this report we describe the cloning and expression pattern of a novel member of this gene family that is specifically expressed in the developing and adult nervous system and thus was designated brain (B)-FABP. B-FABP is closely related to heart (H)-FABP with 67% amino acid identity. B-FABP expression was first detected at mouse embryonic day 10 in neuroepithelial cells and its pattern correlates with early neuronal differentiation. Upon further development, B-FABP was confined to radial glial cells and immature astrocytes. B-FABP mRNA and protein were found in glial cells of the peripheral nervous system such as satellite cells of spinal and cranial ganglia and ensheathing cells of the olfactory nerve layer from as early as embryonic day 11 until adulthood. In the adult mouse brain, B-FABP was found in the glia limitans, in radial glial cells of the hippocampal dentate gyrus and Bergman glial cells. These findings suggest a function of B-FABP during neurogenesis or neuronal migration in the developing nervous system. The partially overlapping expression pattern with that of cellular retinoid binding proteins suggests that B-FABP is involved in the metabolism of a so far unknown hydrophobic ligand with potential morphogenic activity during CNS development.

Amino Acid Sequence↗
PubMed1994

Evidence for radical species as intermediates in cadmium/zinc-metallothionein-dependent DNA damage in vitro.

Toxicologic data on cadmium (Cd) indicate that intracellular metallothionein (MT) is protective for Cd exposure, whereas extracellular Cd-containing MT might be toxic. Moreover, Cd is suspected to be a carcinogen though the underlying mechanism is not known. Here we report on the genotoxic activity of cadmium/zinc-metallothionein (Cd/Zn-MT) in a cell-free test system: a concentration-dependent increase in DNA strand breaks was detected with increasing doses of Cd/Zn-MT, whereas no DNA strand breaks were observed in the presence of heat-denatured MT or Cd or Zn ions alone. Modifications of native Cd/Zn-MT by the metal ion-chelating agent EDTA or the sulfhydryl group alkylating agents N-ethylmaleimide and iodoacetamide suggest that the various cysteine residues of MT, together with the attached heavy metal ions, may be involved in the DNA cleavage reaction. Furthermore, DNA strand breaks caused by Cd/Zn-MT seem more likely to be random than sequence- or base-specific. Results from experiments with radical scavengers and electron spin resonance spectroscopy point to radical species formed by Cd/Zn-MT as mediators of the DNA damage. Thus, the actual activity of Cd/Zn-MT--whether protective or damaging--appears to depend on various parameters governed by the extra- and intracellular environment.

Animals↗
PubMed1994

Supravital uptake of cationic dyes by mast cell granules: a light and electron microscope study.

Methylene blue and neutral red were selected for staining mast cell granules by supravital injections. A new technique was applied for embedding in paraffin and Araldite without dislocation or loss of dye. Stabilization and electron microscopic identification of the dyes were achieved by transforming them into electron-dense precipitates using phosphomolybdic acid dissolved in a paraformaldehyde-glutaraldehyde mixture to preserve the ultrastructure of the tissues. It was found that in general the intensity of the light microscopic staining correlated directly with the electron density. Closer study revealed that not all cytoplasmic granules exhibited the same strong affinity for the cationic dyes. Furthermore, differences in dye distribution were observed within the granules themselves. The difference in the staining pattern can be explained by the heterogeneous occurrence of the anionic residues. Because of its high sensitivity and relatively low toxicity, the method described here is well suited for detecting the binding sites of organic cations in tissues under supravital or vital conditions.

Animals↗
PubMed1994

Large nerve cells with long axons in the granular layer and white matter of the murine cerebellum.

The murine cerebellum was investigated by light microscopy using an improved modification of Ehrlich's methylene blue supravital staining technique. The dye exhibited a special affinity for the perikarya as well as the axons of Purkinje cells. In addition, large fusiform or stellate nerve cells which were characterised by long descending axons were seen to be distributed diffusely within the granular layer and the subcortical white matter. These findings indicate the existence of a 2nd type of projection neuron besides the Purkinje cells and are therefore in full accordance with older neuroanatomical observations based on silver impregnation. When correlated with recent studies on the occurrence of different calcium-binding proteins, the results show that the large perikarya demonstrated immunohistochemically within the granular layer seem to belong to the group of methylene blue positive neurons. Nevertheless, the definitive association of a single neuron with a nerve cell class is only possible if the axon is stained and clearly identifiable. Because of its selectivity for a special type of nerve cell, including its axon, the histological method used in this study may therefore also be suitable for investigating other parts of the brain and the spinal cord.

Animals↗
PubMed1994

[Change in self-object differentiation in an eating disordered patient during inpatient therapy].

This paper is intended as a contribution to qualitative-quantitative research on the psychotherapeutic process. It reports on the course of a three-month inpatient psychosomatic treatment of a female patient with an eating disorder from the preferred point of view of psychoanalytically oriented individual therapy. Given the central role of the self-object relationship in women with eating disorders, changes that occur with regard to this aspect over the course of the therapy are recorded and described. In addition to the clinical perspective, the selected approach is based on a combination of methods, with an emphasis on the analysis of linguistic content using the Gottschalk-Gleser method, the Core Conflictual Relationship Theme method according to Luborsky and the projective procedure of Object Relations Technique according to Phillipson. In the course of treatment reveals changes in terms of reducing shame-anxiety and inwardly-directed aggressiveness as well as in the patient's object repproachement and to a more positive way of dealing with herself, all of which, on the whole, can be considered indicative of an increasing degree of self-object differentiation.

Adult↗
PubMed1994

[Emergency intracoronary stent implantation: complications and experiences with 124 patients].

From January 1990 to February 1993, 88 patients (group 1) received an emergency stent implantation with threatening vascular occlusion within the framework of an elective PTCA or a second emergency PTCA after up to 5 days following a primary successful PTCA. In addition, 36 patients (group 2) received an intracoronary stent during emergency PTCA of instable angina pectoris or acute myocardial infarction. The attempted stent implantation was not successful in 6 other patients. All patients were anticoagulated with heparin, aspirin (100 to 500 mg), and phenprocoumon. Since October 1991, 3 x 75 mg dipyridamole was given and heparinization was stopped after measuring the anticoagulation factor II (prothrombin time < 40%). Main complications within the first 2 to 3 weeks were acute and subacute stent thrombosis (21.8%) and complications of the puncture site (bleeding 19.3%, a. spurium/av-fistula 1.6%). The risk of acute stent thrombosis was significantly higher in patients of group 2 (instable angina pectoris despite of drug therapy or acute myocardial infarction) compared with group 1 (42.4 versus 14.8%). Implantation of multiple stents to stabilize extended dissections had a lower occlusion rate (6.3%). Acute myocardial infarctions were registered in group 1 in 25% (11.4% following implantation, 13.6% following stent occlusion, CK 153 to 3380 U/I, average 826 U/I) and in 58.3% of the high risk patients in group 2 (50% just before or following implantation, 36.1% infarctions or re-infarctions caused by stent occlusion, CK 152 to 1950 U/I, average 657 U/I). The risk of infarction could be limited to approximately 58% of the patients of group 2.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1994

[Prevalence of antibodies against the viruses of European swine fever, Aujeszky's disease and "porcine reproductive and respiratory syndrome" in wild boars in the federal states Sachsen-Anhalt and Brandenburg].

During the hunting season from 1991/1992 blood samples were collected from wild boar shot in the Federal States of Sachsen-Anhalt (482 samples) and Brandenburg (177 samples) which corresponds to 2.1 and 0.4% of the total hunting bag. All sera were screened in a complex trapping blocking (CTB) ELISA for antibodies against hog cholera virus (HCV) and in an indirect ELISA for antibodies against Aujeszky's disease virus (ADV). Additionally the sera were tested for neutralizing antibodies against HCV strain ALFORT/187, bovine viral diarrhoea virus (BVDV) strains NADL and 1138/69, and against an ADV field isolate. In case of questionable results sera were tested against HCV strain "BERGEN", HCV vaccine strain "RIEMS" and three HCV field isolates from wild boar. The serological testing for antibodies against "porcine reproductive and respiratory syndrome virus" (PRRSV) was carried out in indirect immunoperoxidase monolayer assay (IPMA). Four sera (Sachsen-Anhalt) reacted positive in CTB-ELISA. Seven sera yielded neutralizing antibodies against HCV but only one of the "non-negative" samples scored positive in both techniques, ELISA and VNT. Two sera (Brandenburg) had low neutralizing antibody titres against Alfort/197 but scored negative in CTB-ELISA. Screening for antibodies against ADV of 640 sera led to 13 positive sera including 5 positive findings in both ELISA and VNT. Antibodies against PRRSV were detected in two sera which were collected Sachsen-Anhalt. Estimations resulted in a prevalence of about 5% for antibodies against HCV.

Animals↗
PubMed1994

Developmental regulation of voltage-gated K+ channel and GABAA receptor expression in Bergmann glial cells.

Bergmann glial cells are closely associated with neurons: during development they provide guiding structures for migrating granule cells and in the adult cerebellum they display intimate interactions with Purkinje cells. In this study, we have addressed the question of whether such changes in neuronal-glial interactions during development are accompanied by variations in the membrane properties of Bergmann glial cells. We used a mouse cerebellum slice preparation to study membrane currents of the Bergmann glial cells at various stages of development in situ using the patch-clamp technique. The distinct morphology of Bergmann glial cells was revealed by Lucifer yellow injections during recording. While Bergmann glial cells in mice of postnatal day 20 (P20) to P30 have thick processes with arborized, irregularly shaped leaf-like appendages, the processes of cells from younger mice (P5-P7) are thinner and smoother. This morphological maturation is accompanied by a variation in voltage-gated currents. In cells from P5 to P7, delayed outward- and inward-rectifying K+ currents were recorded, while older Bergmann glial cells were characterized by, large, voltage- and time-independent K+ currents. In addition, application of GABA induces two effects, a rapid activation of a Cl- conductance and a longer-lasting decrease in the (resting) K+ conductance. Both effects were mediated by benzodiazepine-insensitive GABAA receptors. Responses in cells of P5-P7 mice were large as compared to the small or even undetectable responses in P20-P30 cells. These GABAA receptors were characterized immunohistochemically in mice and rat brain sections with five subunit-specific antibodies. Bergmann glial cells exhibit a distinct but transient immunoreactivity for the GABAA receptor alpha 2-, alpha 3-, and delta-subunits. Staining is maximal between P7 and P10 and decreases gradually thereafter. In contrast, antibodies to the alpha 1- and beta 2,3-subunits fail to decorate Bergmann glial cells, although they yield a prominent staining of both the Purkinje cells and the granule cells. These changes in the Bergmann glial cell membrane properties and GABAA receptor expression suggest a transition between functional states during development of the Bergmann glial cells.

Aging↗
PubMed1993

Two distinct functional sites of human interleukin 4 are identified by variants impaired in either receptor binding or receptor activation.

Interleukin 4 (IL-4) exerts a decisive role in the coordination of protective immune responses against parasites, particularly helminths. A disregulation of IL-4 function is possibly involved in the genesis of allergic disease states. The search for important amino acid residues in human IL-4 by mutational analysis of charged invariant amino acid positions identified two distinct functional sites in the 4-helix-bundle protein. Site 1 was marked by amino acid substitutions of the glutamic acid at position 9 in helix A and arginine at position 88 in helix C. Exchanges at both positions led to IL-4 variants deficient in binding to the extracellular domain of the IL-4 receptor (IL-4R(ex)). In parallel, up to 1000-fold increased concentrations of this type of variant were required to induce T-cell proliferation and B-cell CD23 expression. Site 2 was marked by amino acid exchanges in helix D at positions 121, 124 and 125 (arginine, tyrosine and serine respectively in the wild-type). IL-4 variants affected at site 2 exhibited partial agonist activity during T-cell proliferation; however, they still bound with high affinity to IL-4R(ex). [The generation of an IL-4 antagonist by replacing tyrosine 124 with aspartic acid has been described before by Kruse et al. (1992) (EMBO J., 11, 3237-3244)]. These findings indicate that IL-4 functions by binding IL-4R(ex) via site 1 which is constituted by residues on helices A and C.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1993

Structure and organisation of a murine gene encoding small heat-shock protein Hsp25.

The structure and sequence of a gene encoding the mouse small heat-shock protein, Hsp25, is presented and compared to the human hsp27. In contrast to the human hsp27, only two copies of hsp25 could be detected in the mouse genome. The intron-exon structure of the identified hsp25 is similar to the transcribed human hsp27, and the transcription start points of the genes are located at similar sites. The promoter region contains various putative transcription factor-binding elements including two G + C-rich Sp1-binding domains, two heat-shock elements, and an estrogen-responsive element half-site in direct proximity to the TATA box. These elements could explain hsp25 basal expression as well as its induction as a result of heat-shock and estrogen treatment.

Amino Acid Sequence↗