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Contains fulltext : 191265.pdf (publisher's version ) (Open Access), MicroRNAs (miRNAs) are involved in post-transcriptional modulation of gene expression and thereby have a large influence on the resulting phenotype. We have previously shown that miRNAs may be involved in the communication between Toxoplasma gondii and its hosts and further confirmed a number of proposed specific miRNAs. Yet, little is known about the internal regulation via miRNAs in T. gondii. Therefore, we predicted pre-miRNAs directly from the type II ME49 genome and filtered them. For the confident hairpins, we predicted the location of the mature miRNAs and established their target genes. To add further confidence, we evaluated whether the hairpins and their targets were co-expressed. Such co-expressed miRNA and target pairs define a functional interaction. We extracted all such functional interactions and analyzed their differential expression among strains of all three clonal lineages (RH, PLK, and CTG) and between the two stages present in the intermediate host (tachyzoites and bradyzoites). Overall, we found ~65,000 expressed interactions of which ~5,500 are differentially expressed among strains but none are significantly differentially expressed between developmental stages. Since miRNAs and target decoys can be used as therapeutics we believe that the list of interactions we provide will lead to novel approaches in the treatment of toxoplasmosis.

Contains fulltext : 191265.pdf (publisher's version ) (Open Access), MicroRNAs (miRNAs) are involved in post-transcriptional modulation of gene expression and thereby have a large influence on the resulting phenotype. We have previously shown that miRNAs may be involved in the communication between Toxoplasma gondii and its hosts and further confirmed a number of proposed specific miRNAs. Yet, little is known about the internal regulation via miRNAs in T. gondii. Therefore, we predicted pre-miRNAs directly from the type II ME49 genome and filtered them. For the confident hairpins, we predicted the location of the mature miRNAs and established their target genes. To add further confidence, we evaluated whether the hairpins and their targets were co-expressed. Such co-expressed miRNA and target pairs define a functional interaction. We extracted all such functional interactions and analyzed their differential expression among strains of all three clonal lineages (RH, PLK, and CTG) and between the two stages present in the intermediate host (tachyzoites and bradyzoites). Overall, we found ~65,000 expressed interactions of which ~5,500 are differentially expressed among strains but none are significantly differentially expressed between developmental stages. Since miRNAs and target decoys can be used as therapeutics we believe that the list of interactions we provide will lead to novel approaches in the treatment of toxoplasmosis.

Contains fulltext : 191265.pdf (publisher's version ) (Open Access), MicroRNAs (miRNAs) are involved in post-transcriptional modulation of gene expression and thereby have a large influence on the resulting phenotype. We have previously shown that miRNAs may be involved in the communication between Toxoplasma gondii and its hosts and further confirmed a number of proposed specific miRNAs. Yet, little is known about the internal regulation via miRNAs in T. gondii. Therefore, we predicted pre-miRNAs directly from the type II ME49 genome and filtered them. For the confident hairpins, we predicted the location of the mature miRNAs and established their target genes. To add further confidence, we evaluated whether the hairpins and their targets were co-expressed. Such co-expressed miRNA and target pairs define a functional interaction. We extracted all such functional interactions and analyzed their differential expression among strains of all three clonal lineages (RH, PLK, and CTG) and between the two stages present in the intermediate host (tachyzoites and bradyzoites). Overall, we found ~65,000 expressed interactions of which ~5,500 are differentially expressed among strains but none are significantly differentially expressed between developmental stages. Since miRNAs and target decoys can be used as therapeutics we believe that the list of interactions we provide will lead to novel approaches in the treatment of toxoplasmosis.

Since the last decade, the interest has risen in nanoscaled technological products, which have advantages through their size effect. The size effect also plays a significant role in the area of micro- and nanoscale heat transfers. Many applications were developed using this effect, such as nanostructured porous media, e.g. Aerogels or ceramics. This experimental work is focused on the determination of thermal accommodation coefficients (TAC) on ceramic surfaces considering several influencing factors. TAC is influenced by temperature, kind of gas, kind of wall material, roughness, and contamination with adsorbed gas layers. To determine TAC, a parallel plate apparatus is used, although most of the measurements in the past have been arranged with the hot wire method. Using the parallel plates apparatus is advantageous due to its simpler machining of specimens and to analyze the effect of roughness. The apparatus is build similar to the well-known guarded hot-plate method using an unidirectional heat flux through the gas layer due to a small temperature gradient, which is provided by two different heat foils. The measurements take place between 30 and 100°C, 10−4, and 1 mbar.

Since the last decade, the interest has risen in nanoscaled technological products, which have advantages through their size effect. The size effect also plays a significant role in the area of micro- and nanoscale heat transfers. Many applications were developed using this effect, such as nanostructured porous media, e.g. Aerogels or ceramics. This experimental work is focused on the determination of thermal accommodation coefficients (TAC) on ceramic surfaces considering several influencing factors. TAC is influenced by temperature, kind of gas, kind of wall material, roughness, and contamination with adsorbed gas layers. To determine TAC, a parallel plate apparatus is used, although most of the measurements in the past have been arranged with the hot wire method. Using the parallel plates apparatus is advantageous due to its simpler machining of specimens and to analyze the effect of roughness. The apparatus is build similar to the well-known guarded hot-plate method using an unidirectional heat flux through the gas layer due to a small temperature gradient, which is provided by two different heat foils. The measurements take place between 30 and 100°C, 10−4, and 1 mbar.

We demonstrate a new instrument for in situ detection of atmospheric iodine atoms and molecules based on atomic and molecular resonance and off-resonance ultraviolet fluorescence excited by lamp emission. The instrument combines the robustness, light weight, low power consumption and efficient excitation of radio-frequency discharge light sources with the high sensitivity of the photon counting technique. Calibration of I-2 fluorescence is achieved via quantitative detection of the molecule by Incoherent Broad Band Cavity-enhanced Absorption Spectroscopy. Atomic iodine fluorescence signal is calibrated by controlled broad band photolysis of known I-2 concentrations in the visible spectral range at atmospheric pressure. The instrument has been optimised in laboratory experiments to reach detection limits of 1.2 pptv for I atoms and 13 pptv for I-2, for S/N = 1 and 10 min of integration time. The ROFLEX system has been deployed in a field campaign in northern Spain, representing the first concurrent observation of ambient mixing ratios of iodine atoms and molecules in the 1-350 pptv range.

Gas-phase heat transfer plays a critical role in many high temperature applications, such as preheaters, combustors, and other thermal equipment. In such cases common heat transfer augmentation methods rely on the convective component alone to achieve improved internal performance. Radiatively assisted heat transfer augmentation has been suggested as a way to overcome limitations in convective-only enhancement. One example of such a technique is the fiber array insert; thermal radiation emitted by tube walls is captured by a large number of slender fibers, which in turn convect heat to the flowing fluid. Previous numerical studies have indicated that this technique represents a promising enhancement method warranting further investigation. This paper presents results from an experimentally based feasibility study of fiber array inserts for heat transfer augmentation in an externally heated duct. Fibers composed of 140 mu m silicon carbide and 150 mu m stainless steel were assembled in arrays with porosities around 0.98, and were tested for empty-tube Reynolds numbers ranging from 17,500 to 112,500 and wall temperatures from ambient up to 750 degrees C. The arrays cause a significant pressure drop-roughly two orders of magnitude higher than the empty-tube case-but tube-side heat transfer coefficients were improved by up to 100% over the convective-only case in the low flow rate regime. The stainless steel fiber array exhibited similar heat transfer performance as the silicon carbide case, although pressure drop characteristics differed owing to variations in fluid-structure flow phenomena. Pressure drop data were roughly within the range of d'Arcy law predictions for both arrays, and deviations could be explained by inhomogeneities in fiber-to-fiber spacing. Heat transfer was found to depend nonlinearly on wall temperature and flow rate, in contrast to previously reported numerical data., QC 20100525

Immunhistochemische Vergleichsanalyse von Primärzellaggregaten und Ursprungsgeweben unterschiedlicher Dignität zur Charakterisierung der in-vitro Anpassung Zielsetzung: Dreidimensionale Zellkulturen stellen eine Weiterentwicklung der einschichtigen Zellkultur dar, die den in-vitro Bedingungen im Organismus näherkommt. Darüber hinaus ermöglichen organoide Kulturen, die neben der vorherrschenden Tumorzelle auch Bindegewebszellen und Immunzellen enthalten, eine Analyse der Interaktion dieser Zellen bei wichtigen Aspekten der Krebserkrankung wie Metastasierung, Angiogenese oder der Tumorimmunologie. Materialien und Methoden: Operationsresektate von Schilddrüsengeweben, -adenomen und -karzinomen, Ovarialkarzinomen und Sarkomen wurden in Einzelzellsuspensionen überführt. Nach Inkubation im Schüttler wurden innerhalb von 24-48 Stunden Primärzellaggregate gezüchtet, von denen Kryostatschnitte angefertigt wurden. Mit der APAAP-Methode wurden Epithelzell-, Leukozyten-, Makrophagen- und Endothelzellmarker sowie E-Cadherin, die a2-, a4-, a5- und av Integrinkette, IGF-I und EGF Rezeptoren, cerbB2 sowie Cathepsin D immunhistochemisch untersucht. Die Färbungen der Aggregate und der Herkunftsgewebe wurden statistisch mit dem Mann-Whitney Test verglichen. Ergebnisse: Primäraggregate konnten zu 90-100% aus Operationsresektaten kultiviert werden. Epithelzellen, Leukozyten, Makrophagen und Endothelzellen waren im Primäraggregaten und den Herkunftsgeweben ungefähr gleichmäßig vorhanden. Dies gilt auch für E-Cadherin, a4-Integrin, IGF-I und EGF Rezeptoren, cerbB2 und Cathepsin D. Die a2-, a5- und av Integrinkette trat nur in den Primäraggregaten von Schilddrüsengeweben und -adenomen nicht aber in deren Herkunftsgeweben auf, was auf eine de-novo Exprimierung schließen läßt. Schlußfolgerung: Mit der verwendeten Methode ist es relativ einfach möglich, organoide Primäraggregate zu züchten, die ein geeignetes Forschungsobjekt für verschiedene Aspekte der Tumorpathologie darstellen. Die gefund, Immunohistochemical Analysis of primary cell aggregates and their origin tissue of different pathology to evaluate adaptation to in-vitro environment Objective: Three-dimensional cell cultures reflect more closely the in-vitro environment then monolayer cultures. Furthermore, organoid cultures, which contain beside the dominant tumor cell also mesenchymal cells and leucocytes are used to study the interaction of these cells in several aspects of the tumor pathology, such as metastasis, angiogenesis and tumor immunology. Materials and Methods: Specimens obtained from thyroid tissue, thyroid adenomas and carcinomas, ovarian cancer and sarcomas were dissolved to single cell suspensions. After incubation under stirring, primary cell aggregates were cultured within 24-48 hours. Cryostat sections were made and stained with markers of epithelial cells, leucocytes, macrophages, endothelial cells as well as E-cadherin, a2-, a4-, a5- und av Integrin chain, IGF-I und EGF receptor, cerbB2 and Cathepsin D using the APAAP method. The immunhistochemical results of the aggregates and their origin tissue were statistically compared using the Mann-Whitney test. Results: Primary cell aggregates could be obtained from up to 90-100% of all probes. Epithelial cells, leucocytes, macrophages and endothelial cells were found equally in aggregates and their origin tissue. Also E-Cadherin, a4-Integrin, IGF-I und EGF receptors, cerbB2 und Cathepsin D were found equally. The a2-, a5- und av integrin chain was expressed in aggregates of thyroid tissue and adenomas, but not in their origin tissue suggesting a de-novo expression. Conclusion: Primary cell aggregates were easily obtained with the used method and could be used as a model in the study of tumor pathology. The different expressions of integrins show an adaptation to the in-vitro environment and could be a reaction to avoid matrix-related apoptosis.

Immunhistochemische Vergleichsanalyse von Primärzellaggregaten und Ursprungsgeweben unterschiedlicher Dignität zur Charakterisierung der in-vitro Anpassung Zielsetzung: Dreidimensionale Zellkulturen stellen eine Weiterentwicklung der einschichtigen Zellkultur dar, die den in-vitro Bedingungen im Organismus näherkommt. Darüber hinaus ermöglichen organoide Kulturen, die neben der vorherrschenden Tumorzelle auch Bindegewebszellen und Immunzellen enthalten, eine Analyse der Interaktion dieser Zellen bei wichtigen Aspekten der Krebserkrankung wie Metastasierung, Angiogenese oder der Tumorimmunologie. Materialien und Methoden: Operationsresektate von Schilddrüsengeweben, -adenomen und -karzinomen, Ovarialkarzinomen und Sarkomen wurden in Einzelzellsuspensionen überführt. Nach Inkubation im Schüttler wurden innerhalb von 24-48 Stunden Primärzellaggregate gezüchtet, von denen Kryostatschnitte angefertigt wurden. Mit der APAAP-Methode wurden Epithelzell-, Leukozyten-, Makrophagen- und Endothelzellmarker sowie E-Cadherin, die a2-, a4-, a5- und av Integrinkette, IGF-I und EGF Rezeptoren, cerbB2 sowie Cathepsin D immunhistochemisch untersucht. Die Färbungen der Aggregate und der Herkunftsgewebe wurden statistisch mit dem Mann-Whitney Test verglichen. Ergebnisse: Primäraggregate konnten zu 90-100% aus Operationsresektaten kultiviert werden. Epithelzellen, Leukozyten, Makrophagen und Endothelzellen waren im Primäraggregaten und den Herkunftsgeweben ungefähr gleichmäßig vorhanden. Dies gilt auch für E-Cadherin, a4-Integrin, IGF-I und EGF Rezeptoren, cerbB2 und Cathepsin D. Die a2-, a5- und av Integrinkette trat nur in den Primäraggregaten von Schilddrüsengeweben und -adenomen nicht aber in deren Herkunftsgeweben auf, was auf eine de-novo Exprimierung schließen läßt. Schlußfolgerung: Mit der verwendeten Methode ist es relativ einfach möglich, organoide Primäraggregate zu züchten, die ein geeignetes Forschungsobjekt für verschiedene Aspekte der Tumorpathologie darstellen. Die gefund, Immunohistochemical Analysis of primary cell aggregates and their origin tissue of different pathology to evaluate adaptation to in-vitro environment Objective: Three-dimensional cell cultures reflect more closely the in-vitro environment then monolayer cultures. Furthermore, organoid cultures, which contain beside the dominant tumor cell also mesenchymal cells and leucocytes are used to study the interaction of these cells in several aspects of the tumor pathology, such as metastasis, angiogenesis and tumor immunology. Materials and Methods: Specimens obtained from thyroid tissue, thyroid adenomas and carcinomas, ovarian cancer and sarcomas were dissolved to single cell suspensions. After incubation under stirring, primary cell aggregates were cultured within 24-48 hours. Cryostat sections were made and stained with markers of epithelial cells, leucocytes, macrophages, endothelial cells as well as E-cadherin, a2-, a4-, a5- und av Integrin chain, IGF-I und EGF receptor, cerbB2 and Cathepsin D using the APAAP method. The immunhistochemical results of the aggregates and their origin tissue were statistically compared using the Mann-Whitney test. Results: Primary cell aggregates could be obtained from up to 90-100% of all probes. Epithelial cells, leucocytes, macrophages and endothelial cells were found equally in aggregates and their origin tissue. Also E-Cadherin, a4-Integrin, IGF-I und EGF receptors, cerbB2 und Cathepsin D were found equally. The a2-, a5- und av integrin chain was expressed in aggregates of thyroid tissue and adenomas, but not in their origin tissue suggesting a de-novo expression. Conclusion: Primary cell aggregates were easily obtained with the used method and could be used as a model in the study of tumor pathology. The different expressions of integrins show an adaptation to the in-vitro environment and could be a reaction to avoid matrix-related apoptosis.

The diagnostic performance of single-serum assays for toxoplasma-specific immunoglobulin (Ig)M. IgA. IgG, and IgE antibodies and of different combinations of such antibody assays in 20 European reference centers was assessed. A panel of 276 sera, of which 73 came from patients who seroconverted within 3 months (acute infection), 49 from patients who had seroconverted 3-12 months earlier (convalescence), and 154 from subjects who had two IgG-positive samples obtained 12 months apart (past infection), was tested with 20 toxoplasma-antibody assays and 195 combinations. In general, every assay with high diagnostic sensitivity showed low diagnostic specificity, i.e. no assay performed alone could reliably distinguish acute from past infection. Furthermore, no single assay (or combination) could separate convalescence from the other stages of toxoplasma infection. However, excellent diagnostic performances were reached by sequential use of highly sensitive IgM assays and methods examining IgG avidity or stage specificity. IgA or IgM assays were less suitable for confirmation of toxoplasma-IgM positivity. This study documents the strength of test combinations in assessing the stage of toxoplasma infection.

The technique evolved for solidifying liquid hydrogen and neon to the desired density is described herein as are the evaluation of a prototype model pressure-control valve and completion of the deliverable model of the cryogenic- solid cooler.