Biomedical subjects

Stefanie Müller

Publications and source records attributed to Stefanie Müller.

7 recordsLinked to original sources

PubMed2006

Mesenchymal stem cells display coordinated rolling and adhesion behavior on endothelial cells.

To explore the initial steps by which transplanted mesenchymal stem cells (MSCs) interact with the vessel wall in the course of extravasation, we studied binding of human MSCs to endothelial cells (ECs). In a parallel plate flow chamber, MSCs bound to human umbilical vein ECs (HUVECs) similar to peripheral-blood mononuclear cells (PBMCs) or CD34(+) hematopoietic progenitors at shear stresses of up to 2 dynes/cm(2). This involved rapid extension of podia, rolling, and subsequent firm adhesion that was increased when ECs were prestimulated with TNF-alpha. MSC binding was suppressed when ECs were pretreated with function-blocking anti-P-selectin antibody, and rolling of MSCs was induced on immobilized P-selectin, indicating that P-selectin was involved in this process. Preincubation of HUVECs with anti-VCAM-1 or of MSCs with anti-VLA-4 antibodies suppressed binding of MSCs to HUVECs but did not enhance inhibition by anti-P-selectin, indicating that both P-selectin and VCAM-1 are equally required for this process. Intravital microscopy demonstrated the capacity of MSCs to roll and adhere to postcapillary venules in vivo in a mouse model in a P-selectin-dependent manner. Thus, MSCs interact in a coordinated fashion with ECs under shear flow, engaging P-selectin and VCAM-1/VLA-4.

Animals↗

PubMed2006

Regulation of expression of atypical orphan response regulators of Helicobacter pylori.

The human gastric pathogen Helicobacter pylori exhibits a remarkably small repertoire of transcriptional regulators including three complete two-component systems as well as the orphan response regulators HP1021 and HP1043. Both HP1021 and HP1043 show atypical receiver sequences and are required for the normal cell growth of H. pylori. Recently, we demonstrated that phosphorylation of HP1021 and HP1043 according to the two-component paradigm is not a prerequisite for the cell growth-associated functions of these response regulators, raising the question of how the activity of this regulatory proteins is modulated. Here, we report that strict transcriptional control of its expression is not involved in the cell-growth associated function of HP1021. We show that expression of hp1043 is controlled both on the post-transcriptional or post-translational level and by transcriptional regulation. Furthermore, we provide evidence that hp1043 can be replaced by the orthologous gene cj0355 from Campylobacter jejuni.

Adaptation, Physiological↗

PubMed2006

Dinuclear alkylamine platinum(II) complexes of [1,2-bis(4-fluorophenyl)ethylenediamine]platinum(II): influence of endocytosis and copper and organic cation transport systems on cellular uptake.

Various possible pathways for the uptake of cationic alkylamine platinum(II) complexes into the MCF-7 breast cancer cells were studied with di[meso-1,2-bis(4-fluorophenyl)ethylenediamine]di[sulfinylbis(methane)-S][mu-1,6-diaminohexaneN:N']diplatinum(II) disulfate (m-4F-PtDMSO-DAH) as an example. It was demonstrated that m-4F-PtDMSO-DAH competed neither for the copper transporter nor the organic cation transporters (OCT and OATP). Instead, adsorptive endocytosis by macropinocytosis played an essential role. Inhibitors of this processes such as amiloride, N-ethyl-N-isopropylamiloride (EIPA), wortmannin, and cytochalasin D decreased the intracellular uptake of m-4F-PtDMSO-DAH dramatically. These results support the understanding of the pharmacological behavior of this promising drug family, which showed no cross resistance with cisplatin.

Amines↗

PubMed2006

Hyperhomocysteinemia--the biochemical link between a weak heart and brittle bones?

Osteoporosis, chronic heart failure (CHF) and mild to moderate hyperhomocysteinemia (HHCY) can frequently be found in elderly individuals and often occur in the same individual. Due to demographic changes in the number of elderly people the total number of individuals suffering from osteoporosis and/or CHF, and hence the cost to society, will increase dramatically over the next 50 years. Thus, prevention of these diseases by identifying and modifying risk factors is a major issue. Recent large population-based prospective studies suggested HHCY as an independent risk factor for CHF and osteoporosis. However, the mechanisms that link HHCY to CHF and osteoporosis are almost unknown. Moreover, until now both diseases have been considered as independent diseases. The finding that heart and bones share a common biochemical risk factor raises the question if there is a biochemical link between these two diseases? This manuscript reviews the existing literature about HHCY and osteoporosis, about HHCY and CHF, and about possible mechanisms that link HHCY to both diseases. Existing data suggest that HHCY could be a useful paradigm with most interesting therapeutic implications, because HCY lowering therapy could favourably influence the course in CHF and osteoporotic patients.

Animals↗

PubMed2005

Repeated sequence motifs in the Helicobacter pylori P1408 promoter do not affect its transcription.

The ArsRS two-component system controls the pH-dependent transcription of several target genes involved in the acid resistance of Helicobacter pylori. In its phosphorylated form the response regulator ArsR activates transcription of the urease genes and it has been reported that ArsR approximately P binds to a 26 bp consensus motif which is present in the promoter regions of the ORFs hp1408, hp119 and hp1432 encoding proteins of unknown function. Here we show that the upstream region of ORF hp1408 exhibits considerable sequence variation in different isolates of H. pylori. By the construction of fusions of the P(1408) promoter from different H. pylori strains to the reporter gene gfp in the genetic background of H. pylori G27 we demonstrate that these sequence variations do not significantly affect acid-induced transcription. Furthermore, we show that a P(1408) core promoter comprising only the -10 promoter element and the 26 bp ArsR binding site overlapping the -35 region is sufficient for eliciting the normal acid response of ORF hp1408.

Bacterial Proteins↗

PubMed2004

Replicon system for Lassa virus.

Lassa virus is endemic to West Africa and causes hemorrhagic fever in humans. To facilitate the functional analysis of this virus, a replicon system was developed based on Lassa virus strain AV. Genomic and antigenomic minigenomes (MG) were constructed consisting of the intergenic region of S RNA and a reporter gene (Renilla luciferase) in antisense orientation, flanked by the 5' and 3' untranslated regions of S RNA. MGs were expressed under the control of the T7 promoter. Nucleoprotein (NP), L protein, and Z protein were expressed from plasmids containing the T7 promoter and internal ribosomal entry site. Transfection of cells stably expressing T7 RNA polymerase (BSR T7/5) with MG in the form of DNA or RNA and plasmids for the expression of NP and L protein resulted in high levels of Renilla luciferase expression. The replicon system was optimized with respect to the ratio of the transfected constructs and by modifying the 5' end of the MG. Maximum activity was observed 24 to 36 h after transfection with a signal-to-noise ratio of 2 to 3 log units. Northern blot analysis provided evidence for replication and transcription of the MG. Z protein downregulated replicon activity close to background levels. Treatment with ribavirin and alpha interferon inhibited replicon activity, suggesting that both act on the level of RNA replication, transcription, or ribonucleoprotein assembly. In conclusion, this study describes the first replicon system for a highly pathogenic arenavirus. It is a tool for investigating the mechanisms of replication and transcription of Lassa virus and may facilitate the testing of antivirals outside a biosafety level 4 laboratory.

Animals↗

PubMed2003

Identification of a novel coronavirus in patients with severe acute respiratory syndrome.

BACKGROUND: The severe acute respiratory syndrome (SARS) has recently been identified as a new clinical entity. SARS is thought to be caused by an unknown infectious agent. METHODS: Clinical specimens from patients with SARS were searched for unknown viruses with the use of cell cultures and molecular techniques. RESULTS: A novel coronavirus was identified in patients with SARS. The virus was isolated in cell culture, and a sequence 300 nucleotides in length was obtained by a polymerase-chain-reaction (PCR)-based random-amplification procedure. Genetic characterization indicated that the virus is only distantly related to known coronaviruses (identical in 50 to 60 percent of the nucleotide sequence). On the basis of the obtained sequence, conventional and real-time PCR assays for specific and sensitive detection of the novel virus were established. Virus was detected in a variety of clinical specimens from patients with SARS but not in controls. High concentrations of viral RNA of up to 100 million molecules per milliliter were found in sputum. Viral RNA was also detected at extremely low concentrations in plasma during the acute phase and in feces during the late convalescent phase. Infected patients showed seroconversion on the Vero cells in which the virus was isolated. CONCLUSIONS: The novel coronavirus might have a role in causing SARS.

Adult↗