Search PubMed⌕ Search

Biomedical subjects

M Steinmetz

Publications and source records attributed to M Steinmetz.

At least 55 records · Page 3Linked to original sources
PubMed1991

Allelic exclusion at DNA rearrangement level is required to prevent coexpression of two distinct T cell receptor beta genes.

In mice double transgenic for functionally rearranged T cell receptor (TCR) V beta 2 and V beta 8.2 genes we found that most T lymphocytes express both TCR beta chains simultaneously. These T cells show no abnormality in thymic selection in vivo and their TCRs are capable of transducing activation signals in vitro. These results indicate that multispecific T cells may appear in the periphery if allelic exclusion of TCR beta genes is not established at the level of gene rearrangement.

Alleles↗
PubMed1991

Dual effect of a Tn917 insertion into the Bacillus subtilis sacX gene.

The most common effect of transposon insertion is the inactivation of genes. However, in some cases, transposons can activate in cis the expression of genes in the neighbourhood of their integration site. We previously described an insertion of the transposon Tn917 into the Bacillus subtilis sacXY locus. sacX and sacY encode respectively a negative and a positive regulator involved in induction by sucrose of the exoenzyme levansucrase. Data in this paper show that the Tn917 insertion had two effects: it inactivated sacX and it increased the transcription of sacY. The latter effect involved one or several elements internal to the transposon.

Bacillus subtilis↗
PubMed1991

Construction of a derivative of Tn917 containing an outward-directed promoter and its use in Bacillus subtilis.

Engineered variants of the transposon Tn917 have been widely used to obtain insertion mutations and transcriptional fusions in Bacillus subtilis and other Gram-positive bacteria. We have developed a novel Tn917-based methodology useful for isolation and characterization of mutants resulting from gene over-expression. A Tn917 variant was constructed which contains a strong out-facing promoter near one end, able to promote transcription of genes in the vicinity of its insertion target. This transposon, designated Tn917PF1, was tested in model conditions. Three Tn917PF1 mutants of B. subtilis, with phenotypes presumed to result from gene over-expression, were analysed. Their phenotypes were shown to be due to transcription from the transposon promoter. In one mutant the promoter activated a deg gene, probably degQ. The other two contained different insertions decryptifying a B. subtilis gene encoding beta-galactosidase.

Bacillus subtilis↗
PubMed1991

Tumor necrosis factor: receptors and inhibitors.

Tumor necrosis factor (TNF) is a highly potent pleiotropic response modifier in inflammatory and immunologic host defense reactions. It can also be toxic to cells and elicit toxic systemic reactions, as evinced by certain pathophysiologic conditions that are initiated or aggravated by an excess of TNF. The cellular mechanisms for transducing TNF signals are complex. There are two forms of TNF, alpha and beta, and two distinct TNF receptors. Many cells express both receptor types simultaneously, even though neither membrane receptor can distinguish between TNF-alpha and TNF-beta. The effects of TNF are inhibited by binding proteins that are truncated fragments of the extracellular domains of the TNF receptors. The mechanisms by which these components of the TNF signal transmission pathways interact to mediate the pleiotropic effects of TNF remain unclear.

Humans↗
PubMed1990

Structure of the murine mb-1 gene encoding a putative sIgM-associated molecule.

Genomic DNA clones containing the B cell-specific murine mb-1 gene were isolated and a 5.6-kb BamH I fragment was characterized. It is 5629 bp long and contains five exons: an exon containing the 5' untranslated and the coding sequence of the signal peptide, an exon of 294 bp, which contains most of the extracellular sequence of the MB-1 protein, a 119-bp long exon coding mainly for the transmembrane portion, and two exons of 69 bp and 427 bp encoding the cytoplasmic domain and the 3'-untranslated region, respectively. The mb-1 gene does not contain a "TATA box" found in many eukaryotic promoters. The 5'-flanking region has sequence stretches homologous to IgVH 5'-promoter regions and a bcl 2 intron sequence. It contains the decanucleotide sequence (ATGGCAAATA) almost identical to the octamer motif of IgVH promoters. A B cell-specific DNase I-hypersensitive site was found in the 3'-flanking region indicating that this region might be involved in B cell-specific expression of mb-1. Southern blot analysis of genomic liver DNA with the cloned mb-1 cDNA suggests the existence of another mb-1-related gene segment.

Amino Acid Sequence↗
PubMed1990

Nucleotide sequence of the sacS locus of Bacillus subtilis reveals the presence of two regulatory genes.

The nucleotide sequence of 3 kb of Bacillus subtilis chromosomal DNA, including sacS, reveals that the regulatory locus for levansucrase synthesis consists of two genes, sacX and sacY. The sacX gene product is remarkably similar to sucrose-specific enzyme II of the B. subtilis phosphoenolpyruvate-dependent phosphotransferase system. The product of sacY is similar, both in amino acid sequence and most probably in its function, to BglG, an Escherichia coli transcriptional antitermination factor.

Amino Acid Sequence↗
PubMed1990

T-cell specific rearrangement of T-cell receptor beta transgenes in mice.

To study rearrangement of T cell receptor (TCR) genes, transgenic mice were generated with a TCR beta minilocus in germline configuration, containing three V beta, two D beta, fourteen J beta and two C beta gene segments and the TCR beta enhancer. Using the polymerase chain reaction as an analytical tool both partial DJ as well as complete VDJ rearrangements were seen, indicating that the minilocus contained all sequence elements required for rearrangment. Rearrangements of minilocus gene segments were restricted to T cells in the thymus and the periphery and did not occur in B cells. V beta 8.3 and V beta 5 sequences encoded by the minilocus were expressed on the surface of peripheral T cells at high frequencies. Transgenic mice with TCR minilocus genes will be a useful system to identify DNA sequence elements required for regulation of rearrangement in vivo.

Animals↗
PubMed1990

Recombinational hotspot specific to female meiosis in the mouse major histocompatibility complex.

The wm7 haplotype of the major histocompatibility complex (MHC), derived from the Japanese wild mouse Mus musculus molossinus, enhances recombination specific to female meiosis in the K/A beta interval of the MHC. We have mapped crossover points of fifteen independent recombinants from genetic crosses of the wm7 and laboratory haplotypes. Most of them were confined to a short segment of approximately 1 kilobase (kb) of DNA between the A beta 3 and A beta 2 genes, indicating the presence of a female-specific recombinational hotspot. Its location overlaps with a sex-independent hotspot previously identified in the Mus musculus castaneus CAS3 haplotype. We have cloned and sequenced DNA fragments surrounding the hotspot from the wm7 haplotype and the corresponding regions from the hotspot-negative B10.A and C57BL/10 strains. There is no significant difference between the sequences of these three strains, or between these and the published sequences of the CAS3 and C57BL/6 strains. However, a comparison of this A beta 3/A beta 2 hotspot with a previously characterized hotspot in the E beta gene revealed that they have a very similar molecular organization. Each hotspot consists of two elements, the consensus sequence of the mouse middle repetitive MT family and the tetrameric repeated sequences, which are separated by 1 kb of DNA.

Animals↗
PubMed1990

Expression of levansucrase-beta-galactosidase hybrids inhibits secretion and is lethal in Bacillus subtilis.

The lacZ gene of Escherichia coli was fused to several positions downstream from the 5' end of the Bacillus subtilis sacB gene, which encodes levansucrase (LS), a sucrose-inducible extracellular enzyme. Effects of hybrid protein expression in B. subtilis were studied. Several fusions were tested, and two significantly interfered with growth of cells and with LS secretion when induced with sucrose. Chromosomal amplification of the fusions, leading to strong expression of the hybrid proteins, completely blocked LS secretion and was lethal for B. subtilis when expression was induced.

Bacillus subtilis↗
PubMed1990

Model reference adaptive control with constraints for postoperative blood pressure management.

This paper presents an approach for adaptive control of blood pressure using sodium nitroprusside. A modified stochastic model reference adaptive control algorithm with a time varying reference model is developed and used for this purpose. An automatic adjustment of the reference model is proposed in order to optimize the performance of the closed loop system while meeting clinical constraints imposed on the infusion rate and the mean arterial pressure. Extensive computer simulations show the robustness of the proposed controller in the presence of relatively high noise levels, over the full range of plant parameters and for large parameter variations.

Algorithms↗
PubMed1990

Induction of levansucrase in Bacillus subtilis: an antitermination mechanism negatively controlled by the phosphotransferase system.

The target of the induction by sucrose of the levansucrase gene is a transcription terminator (sacRt) located upstream from the coding sequence, sacB. The two-gene locus sacX-sacY (formerly sacS) and the ptsI gene were previously shown to be involved in this induction. ptsI encodes enzyme I of the phosphoenolpyruvate-dependent phosphotransferase system. SacX is strongly homologous to sucrose-specific phosphotransferase system-dependent permeases. SacY is a positive regulator of sacB. Here we show that SacY is probably an antiterminator interacting directly with sacRt, since in Escherichia coli the presence of the sacY gene stimulates the expression of a reporter gene fused downstream from sacRt. Missense mutations affecting sacY were sequenced, and the sacB regulation was studied in isogenic strains carrying these mutations or in vitro-generated mutations affecting sacX, sacY, or ptsI. The phenotype of double mutants suggests a model in which SacX might be a sucrose sensor that would be phosphorylated by the phosphotransferase system and, in this state, could inhibit the SacY antiterminator. Exogenous sucrose, or a mutation inactivating the phosphotransferase system, would dephosphorylate SacX and allow antitermination at sacRt.

Bacillus subtilis↗
PubMed1990

Effect of endobronchial radiation therapy on malignant bronchial obstruction.

We evaluated the effect of endobronchial radiation therapy in 52 patients with malignant airway occlusion. Fifty-five endobronchial applications of the radioisotope iridium 192 were carried out. Response was assessed by change in performance status, symptom resolution, duration of symptom relief, roentgenographic reaeration, pulmonary function tests, and postimplant bronchoscopy. Thirty-three patients showed at least a one-level improvement in performance status. Of a total of 166 symptoms present prior to therapy, 131 resolved or improved. Approximately 70 percent of a patient's lifetime was rendered symptom improved or symptom free. A roentgenographic reaeration response of 30/41 (73 percent) was achieved. The average FEV1 and FVC improved from 1.5 to 2.1 L and from 2.3 to 2.9 L, respectively. Posttherapy bronchoscopy was performed between one and two months following the implant in 15 patients who agreed to undergo the procedure. Eleven (73 percent) of 15 had complete tumor regression. Major long-term complications were noted in seven patients. Endobronchial radiation, therefore, appears to be a safe and effective technique to palliate malignant airway occlusion.

Aged↗
PubMed1989

Phosphoenolpyruvate:sugar phosphotransferase system of Bacillus subtilis: nucleotide sequence of ptsX, ptsH and the 5'-end of ptsI and evidence for a ptsHI operon.

The nucleotide sequence of a 1689bp fragment of the Bacillus subtilis locus containing ptsX (a crr-like gene), ptsH (coding for HPr), and the 5'-end of ptsI (coding for Enzyme I) was determined. The deduced amino acid sequences of ptsH and the N-terminal part of ptsI were compared to those of Streptococcus faecalis and Escherichia coli. Transcription fusion demonstrated that ptsHI constitutes an operon. An open reading frame overlapping the main part of ptsH in the opposite sense was shown to be expressed in vivo, using protein fusions with beta-galactosidase. The deduced amino acid sequence of ptsX showed significant homology with that of Salmonella typhimurium glucose-specific Enzyme III. ptsX was preceded by an open reading frame whose amino acid sequence showed strong homology with the C-terminal part of E. coli Enzyme IIGlc.

Amino Acid Sequence↗
PubMed1989

Induction of saccharolytic enzymes by sucrose in Bacillus subtilis: evidence for two partially interchangeable regulatory pathways.

Sucrose induces two saccharolytic enzymes in Bacillus subtilis, an intracellular sucrase and an extracellular levansucrase, encoded by sacA and sacB, respectively. It was previously shown that the sacY gene encodes a positive regulator involved in a sucrose-dependent antitermination upstream from the sacB coding sequence. We show here that the sacY product is not absolutely required for sacB induction: a weak but significant induction can be observed in strains harboring a sacY deletion. The sacY-independent induction was altered by mutations located in the sacP and sacT loci but was observed in both sacU+ and sacU32 genetic backgrounds. These results suggest that B. subtilis has two alternative systems allowing sacB induction by sucrose. Both systems also seem to be involved in sacA induction.

Amino Acid Sequence↗
PubMed1989

Transgenic mice to study T-cell receptor gene regulation and repertoire formation.

Transgenic mice have been obtained with genes coding for an alpha beta T-cell receptor that recognizes the male-specific antigen H-Y in association with the Db class I major histocompatibility complex molecule. Most if not all of the T-cells express the beta chain encoded by the transgene and show allelic exclusion of endogenous beta genes. In contrast, the expression of the alpha transgene does not completely block rearrangement and formation of functional endogenous alpha genes. In H-2b transgenic female mice the transgenic T-cell receptor is functionally expressed on at least 30% of CD8+ peripheral T-lymphocytes as indicated by their ability to lyse male target cells. Also in transgenic H-2b male mice a large proportion of peripheral T-cells appear to express the transgenic receptor. However, these cells do not react with male target cells because they show only low level or no expression of CD8 cell interaction molecules. Tolerance is established in the male transgenic thymus through deletion of CD4+CD8+ immature thymocytes.

Animals↗
PubMed1989

[Trans-scleral ciliary body coagulation with the neodymium:YAG-cw-laser in secondary glaucoma].

Transscleral coagulation of the ciliary body was performed by means of the continuous wave Neodymium: YAG laser in 39 eyes with various types of secondary glaucoma that was not controlled by medical therapy. Three days later, the mean intraocular pressure measured 54% of the pre-laser pressure; 3 months later it was 52%, 6 months later 47%, and 12 months later 58%. In 11 of 14 eyes (78.6%) treated in an attempt to preserve the remaining visual function, the pressure proved to be under control (less than or equal to 20 mmHg) 6 months after treatment. Major complications, such as hyphema or bleeding into the vitreous, were rare (7/39) and occurred only in the eyes with neovascular glaucoma.

Ciliary Body↗
PubMed1988

T-cell-specific deletion of T-cell receptor transgenes allows functional rearrangement of endogenous alpha- and beta-genes.

In B cells the loci encoding immunoglobulin chains usually show allelic exclusion; a given B cell transcribes and translates only one productively rearranged allele of the heavy and light chain loci. This ensures that each B cell expresses only one antigen receptor. The loci encoding T-cell receptor (TCR) alpha- and beta-genes may behave similarly. We have previously reported that the expression of a transgenic TCR beta-chain prevents functional and nonfunctional V beta rearrangements in the endogenous beta-chain loci but not D beta J beta rearrangements. We have also been unable to detect the expression of the TCR gamma-chain locus in thymocytes of these mice (unpublished observations). To study the mechanisms involved in forming a mature T-cell repertoire further, we have constructed mice expressing alpha- and beta-TCR transgenes derived from a cytotoxic T-cell clone that is specific for the male antigen H-Y in the context of H-2Db MHC molecules. Here we show that in these mice rearrangement of endogenous alpha-chain loci is also suppressed, although to a lesser extent than rearrangement of beta-chain loci. In addition, in male alpha beta TCR transgenic mice we observed T-cell clones which had deleted both transgenic alpha- and beta-chain genes and expressed endogenous alpha- and beta-chain TCR genes. These cells are presumably derived from rare thymocytes that leave the male thymus because their TCR no longer recognizes self antigen. The vast majority of CD4+8+ nonmature thymocytes expressing alpha- and beta-transgenes are deleted in the male thymus.

Animals↗