Activation of G protein-coupled receptors : the role of extracellular loops in adenosine receptors

The research described in this thesis has provided new insights in the activation mechanism of class A GPCRs and in particular of adenosine receptors. By a variety of mutagenesis approaches and the use of a robust yeast reporter gene system, we identified several regions and amino acid positions that contribute to both agonist responses and constitutive activity of the human adenosine A1 receptor and the human adenosine A2B receptor. These results reveal new and surprising roles of the extracellular loops in the activation mechanism, greatly contributing to our notion of receptor activation.The research described in this thesis was part of the TI-Pharma initiative "The GPCR forum for established targets" (D1-105)UBL - phd migration 201

Dynamic transitions between metastable states in a superconducting ring

Applying the time-dependent Ginzburg-Landau equations, transitions between metastable states of a superconducting ring are investigated in the presence of an external magnetic field. It is shown that if the ring exhibits several metastable states at a particular magnetic field, the transition from one metastable state to another one is governed by both the relaxation time of the absolute value of the order parameter tau_{|psi|} and the relaxation time of the phase of the order parameter tau_{phi}. We found that the larger the ratio tau_{|psi|}tau_{phi} the closer the final state will be to the absolute minimum of the free energy, i.e. the thermodynamic equilibrium. The transition to the final state occurs through a subsequent set of single phase slips at a particular point along the ring.Comment: 7 pages, 6 figures, Revtex 4.0 styl

CFU-S(11) activity does not localize solely with the aorta in the aorta-gonad-mesonephros region

The aorta-gonad-mesonephros (AGM) region is a potent hematopoietic site in the midgestation mouse conceptus and first contains colony-forming units-spleen day 11 (CFU-S(11)) at embryonic day 10 (E10). Because CFU-S(11) activity is present in the AGM region before the onset of hematopoietic stem cell (HSC) activity, CFU-S(11) activity in the complex developing vascular and urogenital regions of the AGM was localized. From E10 onward, CFU-S(11) activity is associated with the aortic vasculature, and is found also in the urogenital ridges (UGRs). Together with data obtained from organ explant cultures, in which up to a 16-fold increase in CFU-S(11) activity was observed, it was determined that CFU-S(11) can be increased autonomously both in vascular sites and in UGRs. Furthermore, CFU-S(11) activity is present in vitelline and umbilical vessels. This, together with the presence of CFU-S(11) in the UGRs 2 days before HSC activity, suggests both temporally and spatially distinct emergent sources of CFU-S(11). (Blood. 2000;96:2902-2904

Allogeneic chondrogenically differentiated human bone marrow stromal cells do not induce dendritic cell maturation

Bone marrow stromal cell (BMSC)-mediated endochondral bone formation may be a promising alternative to the current gold standards of autologous bone transplantation, in the development of novel methods for bone repair. Implantation of chondrogenically differentiated BMSCs leads to bone formation in vivo via endochondral ossification. The success of this bone formation in an allogeneic system depends upon the interaction between the implanted constructs and the host immune system. The current study investigated the effect of chondrogenically differentiated human bone marrow stromal cell (hBMSC) pellets on the maturation and function of dendritic cells (DCs) by directly coculturing bone forming chondrogenic hBMSC pellets and immature or lipopolysaccharide (LPS)-matured DCs in vitro. Allogeneic chondrogenic hBMSC pellets did not affect the expression of CD80, CD86, or HLADR on immature or LPS-matured DCs following 24, 48, or 72 hr of coculture. Furthermore, they did not induce or inhibit antigen uptake or migration of the DCs over time. IL-6 was secreted by allogeneic chondrogenic hBMSC pellets in response to LPS-matured DCs. Overall, this study has demonstrated that maturation of immature DCs was not influenced by allogeneic chondrogenic hBMSC pellets. This suggests that allogeneic chondrogenic hBMSC pellets do not stimulate immunogenic responses from DCs in vitro and are not expected to indirectly activate T cells via DCs. For this reason, allogeneic chondrogenic bone marrow stromal cell pellets are promising candidates for future tissue engineering strategies utilising allogeneic cells for bone repair

Frequency-dependent magnetotransport and particle dynamics in magnetic modulation systems

We analyze the dynamics of a charged particle moving in the presence of spatially-modulated magnetic fields. From Poincare surfaces of section and Liapunov exponents for characteristic trajectories we find that the fraction of pinned and runaway quasiperiodic orbits {\em vs}. chaotic orbits depends strongly on the ratio of cyclotron radius to the structure parameters, as well as on the amplitude of the modulated field. We present a complete characterization of the dynamical behavior of such structures, and investigate the contribution to the magnetoconductivity from all different orbits using a classical Kubo formula. Although the DC conductivity of the system depends strongly on the pinned and runaway trajectories, the frequency response reflects the topology of all different orbits, and even their unusual temporal behavior.Comment: Submitted to PRB - 14 figure files - REVTEX tex

PDX1 DNA methylation distinguishes two subtypes of pancreatic neuroendocrine neoplasms with a different prognosis

DNA methylation is a crucial epigenetic mechanism for gene expression regulation and cell differentiation. Furthermore, it was found to play a major role in multiple pathological processes, including cancer. In pancreatic neuroendocrine neoplasms (PNENs), epigenetic deregulation is also considered to be of significance, as the most frequently mutated genes have an important function in epigenetic regulation. However, the exact changes in DNA methylation between PNENs and the endocrine cells of the pancreas, their likely cell-of-origin, remain largely unknown. Recently, two subtypes of PNENs have been described which were linked to cell-of-origin and have a different prognosis. A difference in the expression of the transcription factor PDX1 was one of the key molecular differences. In this study, we performed an exploratory genome-wide DNA methylation analysis using Infinium Methylation EPIC arrays (Illumina) on 26 PNENs and pancreatic islets of five healthy donors. In addition, the methylation profile of the PDX1 region was used to perform subtyping in a global cohort of 83 PNEN, 2 healthy alpha cell and 3 healthy beta cell samples. In our exploratory analysis, we identified 26,759 differentially methylated CpGs and 79 differentially methylated regions. The gene set enrichment analysis highlighted several interesting pathways targeted by altered DNA methylation, including MAPK, platelet-related and immune system-related pathways. Using the PDX1 methylation in 83 PNEN, 2 healthy alpha cell and 3 healthy beta cell samples, two subtypes were identified, subtypes A and B, which were similar to alpha and beta cells, respectively. These subtypes had different clinicopathological characteristics, a different pattern of chromosomal alterations and a different prognosis, with subtype A having a significantly worse prognosis compared with subtype B (HR 0.22 [95% CI: 0.051–0.95], p = 0.043). Hence, this study demonstrates that several cancer-related pathways are differently methylated between PNENs and normal islet cells. In addition, we validated the use of the PDX1 methylation status for the subtyping of PNENs and its prognostic importance

Semiautomated isolation and molecular characterisation of single or highly purified tumour cells from CellSearch enriched blood samples using dielectrophoretic cell sorting

Background: Molecular characterisation of single circulating tumour cells (CTCs) holds considerable promise for predictive biomarker assessment and to explore CTC heterogeneity. We evaluate a new method, the DEPArray system, that allows the dielectrophoretic manipulation and isolation of single and 100% purified groups of CTCs from pre-enriched blood samples and explore the feasibility of their molecular characterisation.Methods:Samples containing known numbers of two cell populations were used to assess cell loss during sample loading. Cultured breast cancer cells were isolated from spiked blood samples using CellSearch CTC and Profile kits. Single tumour cells and groups of up to 10 tumour cells were recovered with the DEPArray system and subjected to transcriptional and mutation analysis.Results:On average, 40% cell loss was observed when loading samples to the DEPArray system. Expected mutations in clinically relevant markers could be obtained for 60% of single recovered tumour cells and all groups of tumour cells. Reliable gene expression profiles were obtained from single cells and groups of up to 10 cells for 2 out of 3 spiked breast cancer cell lines.Conclusion:We describe a semiautomated workflow for the isolation of small groups of 1 to 10 tumour cells from whole blood samples and provide proof of principle for the feasibility of their comprehensive molecular characterisation

Flame bands: CO + O chemiluminescence as a measure of gas temperature

Carbon monoxide flame band emission (CO+O → CO2+hV) in CO2 microwave plasma is quantified by obtaining absolute calibrated emission spectra at various locations in the plasma afterglow while simultaneously measuring gas temperatures using rotational Raman scattering. Comparison of our results to literature reveals a contribution of O2 Schumann-Runge UV emission at T &gt; 1500 K. This UV component likely results from the collisional exchange of energy between CO2(1B) and O2. Limiting further analysis to T &lt; 1500 K, we demonstrate the utility of CO flame band emission by analyzing afterglows at different plasma conditions. We show that the highest energy efficiency for CO production coincides with an operating condition where very little heat has been lost to the environment prior to ∼3 cm downstream, while simultaneously, T ends up below the level required to effectively freeze in CO. This observation demonstrates that, in CO2 plasma conversion, optimizing for energy efficiency does not require a sophisticated downstream cooling method.</p

Plasma Driven Exsolution for Nanoscale Functionalization of Perovskite Oxides

Perovskite oxides with dispersed nanoparticles on their surface are considered instrumental in energy conversion and catalytic processes. Redox exsolution is an alternative method to the conventional deposition techniques for directly growing well-dispersed and anchored nanoarchitectures from the oxide support through thermochemical or electrochemical reduction. Herein, a new method for such nanoparticle nucleation through the exposure of the host perovskite to plasma is shown. The applicability of this new method is demonstrated by performing catalytic tests for CO2 hydrogenation over Ni exsolved nanoparticles prepared by either plasma or conventional H2 reduction. Compared to the conventional thermochemical H2 reduction, there are plasma conditions that lead to the exsolution of a more than ten times higher Ni amount from a lanthanum titanate perovskite, which is similar to the reported values of the electrochemical method. Unlike the electrochemical method, however, plasma does not require the integration of the material in an electrochemical cell, and is thus applicable to a wide range of microstructures and physical forms. Additionally, when N2 plasma is employed, the nitrogen species are stripping out oxygen from the perovskite lattice, generating a key chemical intermediate, such as NO, rendering this technology even more appealing.</p

Effects of the field modulation on the Hofstadter's spectrum

We study the effect of spatially modulated magnetic fields on the energy spectrum of a two-dimensional (2D) Bloch electron. Taking into account four kinds of modulated fields and using the method of direct diagonalization of the Hamiltonian matrix, we calculate energy spectra with varying system parameters (i.e., the kind of the modulation, the relative strength of the modulated field to the uniform background field, and the period of the modulation) to elucidate that the energy band structure sensitively depends on such parameters: Inclusion of spatially modulated fields into a uniform field leads occurrence of gap opening, gap closing, band crossing, and band broadening, resulting distinctive energy band structure from the Hofstadter's spectrum. We also discuss the effect of the field modulation on the symmetries appeared in the Hofstadter's spectrum in detail.Comment: 7 pages (in two-column), 10 figures (including 2 tables