Calpain-Mediated Processing of Adenylate Cyclase Toxin Generates a Cytosolic Soluble Catalytically Active N-Terminal Domain

Bordetella pertussis, the whooping cough pathogen, secretes several virulence factors among which adenylate cyclase toxin (ACT) is essential for establishment of the disease in the respiratory tract. ACT weakens host defenses by suppressing important bactericidal activities of the phagocytic cells. Up to now, it was believed that cell intoxication by ACT was a consequence of the accumulation of abnormally high levels of cAMP, generated exclusively beneath the host plasma membrane by the toxin N-terminal catalytic adenylate cyclase (AC) domain, upon its direct translocation across the lipid bilayer. Here we show that host calpain, a calcium-dependent Cys-protease, is activated into the phagocytes by a toxin-triggered calcium rise, resulting in the proteolytic cleavage of the toxin N-terminal domain that releases a catalytically active "soluble AC''. The calpain-mediated ACT processing allows trafficking of the "soluble AC'' domain into subcellular organella. At least two strategic advantages arise from this singular toxin cleavage, enhancing the specificity of action, and simultaneously preventing an indiscriminate activation of cAMP effectors throughout the cell. The present study provides novel insights into the toxin mechanism of action, as the calpain-mediated toxin processing would confer ACT the capacity for a space- and time-coordinated production of different cAMP "pools'', which would play different roles in the cell pathophysiology.

Estimating abundances of 0-group western Baltic cod by using pound net fisheries

Nearshore 0-group western Baltic cod are frequently caught as bycatch in the commercial pound net fishery. Pound net fishermen from the Danish Isle of Funen and Lolland and the German Isle of Fehmarn have recorded their catches of small cod between September and December 2008. Abundance patterns were analysed, particularly concerning the influence of abiotic factors (hydrography, meteorology) and the differences between sampling sites. Catch per unit effort (CPUE) differed by site and location, whereas CPUE were highest at Lolland. Correlation between catch and wind/currents were generally weak. However, wind directions and current speeds seem to affect the catch rates. Finally an algorithm was developed to calculate a recruitment index for western Baltic cod recruitment success based on previous analyses.

Saisonale und regionale Unterschiede zwischen den 0-Gruppen-Schollen-Discards in der deutschen Garnelenfischerei

Shrimping in the waddensea is frequently considered to significantly reduce the numbers of juvenile plaice . This investigation aims to reveal the seasonal and regional differences regarding discards in the German waddensea and sets the results in relation to the bycatch projects with German participation. Furthermore, methodological aspects are evaluated to find possible effects on the estimation of the netted numbers. The bycatch problem exists predominantly throughout the summer season, but high catches of juvenile plaice can occur occasionally in autumn. The discard issue seems to be most important in the East Frisian region, while the Elbe and Schleswig-Holstein areas are of lesser importance. Considering the distribution of the fish in the environment is as indispensable as sampling of the shrimp fleet with optimum representativity. There are indices that the contribution of the Dutch shrimp fleet to the discard mortality of juvenile plaice was underestimated in the recent past. A combination of time and area closures as restrictions in combination with the extensive application of selective gears could possibly gain the best protection for the affected species.

Mateda-2.0: Estimation of Distribution Algorithms in MATLAB

This paper describes Mateda-2.0, a MATLAB package for estimation of distribution algorithms (EDAs). This package can be used to solve single and multi-objective discrete and continuous optimization problems using EDAs based on undirected and directed probabilistic graphical models. The implementation contains several methods commonly employed by EDAs. It is also conceived as an open package to allow users to incorporate different combinations of selection, learning, sampling, and local search procedures. Additionally, it includes methods to extract, process and visualize the structures learned by the probabilistic models. This way, it can unveil previously unknown information about the optimization problem domain. Mateda-2.0 also incorporates a module for creating and validating function models based on the probabilistic models learned by EDAs.

Expression and activity profiles of DPP IV/CD26 and NEP/CD10 glycoproteins in the human renal cancer are tumor-type dependent

Background: Cell-surface glycoproteins play critical roles in cell-to-cell recognition, signal transduction and regulation, thus being crucial in cell proliferation and cancer etiogenesis and development. DPP IV and NEP are ubiquitous glycopeptidases closely linked to tumor pathogenesis and development, and they are used as markers in some cancers. In the present study, the activity and protein and mRNA expression of these glycoproteins were analysed in a subset of clear-cell (CCRCC) and chromophobe (ChRCC) renal cell carcinomas, and in renal oncocytomas (RO). Methods: Peptidase activities were measured by conventional enzymatic assays with fluorogen-derived substrates. Gene expression was quantitatively determined by qRT-PCR and membrane-bound protein expression and distribution analysis was performed by specific immunostaining. Results: The activity of both glycoproteins was sharply decreased in the three histological types of renal tumors. Protein and mRNA expression was strongly downregulated in tumors from distal nephron (ChRCC and RO). Moreover, soluble DPP IV activity positively correlated with the aggressiveness of CCRCCs (higher activities in high grade tumors). Conclusions: These results support the pivotal role for DPP IV and NEP in the malignant transformation pathways and point to these peptidases as potential diagnostic markers.

Three-dimensional growth as multicellular spheroid activates the proangiogenic phenotype of colorectal carcinoma cells via LFA-1-dependent VEGF: implications on hepatic micrometastasis

Background: The recruitment of vascular stromal and endothelial cells is an early event occurring during cancer cell growth at premetastatic niches, but how the microenvironment created by the initial three-dimensional (3D) growth of cancer cells affects their angiogenesis-stimulating potential is unclear. Methods: The proangiogenic profile of CT26 murine colorectal carcinoma cells was studied in seven-day cultured 3D-spheroids of <300 mu m in diameter, produced by the hanging-drop method to mimic the microenvironment of avascular micrometastases prior to hypoxia occurrence. Results: Spheroid-derived CT26 cells increased vascular endothelial growth factor (VEGF) secretion by 70%, which in turn increased the in vitro migration of primary cultured hepatic sinusoidal endothelium (HSE) cells by 2-fold. More importantly, spheroid-derived CT26 cells increased lymphocyte function associated antigen (LFA)-1-expressing cell fraction by 3-fold; and soluble intercellular adhesion molecule (ICAM)-1, given to spheroid-cultured CT26 cells, further increased VEGF secretion by 90%, via cyclooxygenase (COX)-2-dependent mechanism. Consistent with these findings, CT26 cancer cells significantly increased LFA-1 expression in non-hypoxic avascular micrometastases at their earliest inception within hepatic lobules in vivo; and angiogenesis also markedly increased in both subcutaneous tumors and hepatic metastases produced by spheroid-derived CT26 cells. Conclusion: 3D-growth per se enriched the proangiogenic phenotype of cancer cells growing as multicellular spheroids or as subclinical hepatic micrometastases. The contribution of integrin LFA-1 to VEGF secretion via COX-2 was a micro environmental-related mechanism leading to the pro-angiogenic activation of soluble ICAM-1-activated colorectal carcinoma cells. This mechanism may represent a new target for specific therapeutic strategies designed to block colorectal cancer cell growth at a subclinical micrometastatic stage within the liver.

Measurement of velocity profiles in a rectangular microchannel with aspect ratio α = 0.35

In this work, we measured 14 horizontal velocity profiles along the vertical direction of a rectangular microchannel with aspect ratio alpha = h/w = 0.35 (h is the height of the channel and w is the width of the channel) using microPIV at Re = 1.8 and 3.6. The experimental velocity profiles are compared with the full 3D theoretical solution, and also with a Poiseuille parabolic profile. It is shown that the experimental velocity profiles in the horizontal and vertical planes are in agreement with the theoretical profiles, except for the planes close to the wall. The discrepancies between the experimental data and 3D theoretical results in the center vertical plane are less than 3.6%. But the deviations between experimental data and Poiseuille's results approaches 5%. It indicates that 2D Poiseuille profile is no longer a perfect theoretical approximation since a = 0.35. The experiments also reveal that, very near the hydrophilic wall (z = 0.5-1 mu m), the measured velocities are significantly larger than the theoretical velocity based on the no-slip assumption. A proper discussion on some physical effects influencing the near wall velocity measurement is given.

Euskarazko denbora-egiturak etiketatzeko gidalerroak v1.0

To interpret the temporal information on texts, a mark-up language that will code that information is needed, in order to make that information automatically reachable. The most used mark-up language is TimeML (Pustejovsky et al., 2003), which has also been choosen for Basque. In this guidelines we present the Basque version of ISO-TimeML (ISO-TimeML working group, 2008). After having analysed the tags, attributes and values created for English, we describe the most appropriate ones to represent Basque time structures’ information.

Influence of HCl on CO and NO emissions in combustion

The influence of HCl on CO and NO emissions was experimentally investigated in an entrained flow reactor (EFR) and an internally circulating fluidized bed (ICFB). The results in EFR show the addition of HCl inhibits CO oxidation and NO formation at 1073 K and 1123 K. At the lower temperature (1073 K) the inhibition of HCl becomes more obvious. In ICFB, chlorine-containing plastic (PVC) was added to increase the concentration of HCl during the combustion of coal or coke. Results show that HCl is likely to enhance the reduction of NO and N2O. HCl greatly increases CO and CH4 emission in the flue gas. A detailed mechanism of CO/NO/HCl/SO2 system was used to model the effect of HCl in combustion. The results indicate that HCl not only promotes the recombination of radicals O, H, and OH, but also accelerates the chemical equilibration of radicals. The influence of HCl on the radicals mainly occurs at 800-1200 K. (C) 2009 Elsevier Ltd. All rights reserved.

Synthesis and potency of long end-associative polymers for mist control

Long linear polymers that are end-functionalized with associative groups were studied as additives to hydrocarbon fluids to mitigate the fire hazard associated with the presence of mist in a crash scenario. These polymers were molecularly designed to overcome both the shear-degradation of long polymer chains in turbulent flows, and the chain collapse induced by the random placement of associative groups along polymer backbones. Architectures of associative groups on the polymer chain ends that were tested included clusters of self-associative carboxyl groups and pairs of hetero-complementary associative units.

Linear polymers with clusters of discrete numbers of carboxyl groups on their chain ends were investigated first: an innovative synthetic strategy was devised to achieve unprecedented backbone lengths and precise control of the number of carboxyl groups on chain ends (N). We found that a very narrow range of N allows the co-existence of sufficient end-association strength and polymer solubility in apolar media. Subsequent steady-flow rheological study on solution behavior of such soluble polymers in apolar media revealed that the end-association of very long chains in apolar media leads to the formation of flower-like micelles interconnected by bridging chains, which trap significant fraction of polymer chains into looped structures with low contribution to mist-control. The efficacy of very long 1,4-polybutadiene chains end-functionalized with clusters of four carboxyl groups as mist-control additives for jet fuel was further tested. In addition to being shear-resistant, the polymer was found capable of providing fire-protection to jet fuel at concentrations as low as 0.3wt%. We also found that this polymer has excellent solubility in jet fuel over a wide range of temperature (-30 to +70°C) and negligible interference with dewatering of jet fuel. It does not cause an adverse increase in viscosity at concentrations where mist-control efficacy exists.

Four pairs of hetero-complementary associative end-groups of varying strengths were subsequently investigated, in the hopes of achieving supramolecular aggregates with both mist-control ability and better utilization of polymer building blocks. Rheological study of solutions of the corresponding complementary associative polymer pairs in apolar media revealed the strength of complementary end-association required to achieve supramolecular aggregates capable of modulating rheological properties of the solution.

Both self-associating and complementary associating polymers have therefore been found to resist shear degradation. The successful strategy of building soluble, end-associative polymers with either self-associative or complementary associative groups will guide the next generation of mist-control technology.

1kHz-0．1TW高效率钛宝石激光放大器

介绍了一种高重复频率掺钛蓝宝石飞秒激光多通高效率放大系统．在抽运功率为23W，入射功率为660mW时，获得7．2W的放大输出，放大效率达30％．经压缩器压缩后，获得单脉冲能量4．5mJ，脉冲宽度为38fs，重复频率为1kHz，峰值功率大于0．1TW的超短超强激光脉冲．

Intramolecular conflict : conformation and self-assembly of architecturally complex macromolecules in solution

The solution behavior of linear polymer chains is well understood, having been the subject of intense study throughout the previous century. As plastics have become ubiquitous in everyday life, polymer science has grown into a major field of study. The conformation of a polymer in solution depends on the molecular architecture and its interactions with the surroundings. Developments in synthetic techniques have led to the creation of precision-tailored polymeric materials with varied topologies and functionalities. In order to design materials with the desired properties, it is imperative to understand the relationships between polymer architecture and their conformation and behavior. To meet that need, this thesis investigates the conformation and self-assembly of three architecturally complex macromolecular systems with rich and varied behaviors driven by the resolution of intramolecular conflicts. First we describe the development of a robust and facile synthetic approach to reproducible bottlebrush polymers (Chapter 2). The method was used to produce homologous series of bottlebrush polymers with polynorbornene backbones, which revealed the effect of side-chain and backbone length on the overall conformation in both good and theta solvent conditions (Chapter 3). The side-chain conformation was obtained from a series of SANS experiments and determined to be indistinguishable from the behavior of free linear polymer chains. Using deuterium-labeled bottlebrushes, we were able for the first time to directly observe the backbone conformation of a bottlebrush polymer which showed self-avoiding walk behavior. Secondly, a series of SANS experiments was conducted on a homologous series of Side Group Liquid Crystalline Polymers (SGLCPs) in a perdeuterated small molecule liquid crystal (5CB). Monodomain, aligned, dilute samples of SGLCP-b-PS block copolymers were seen to self-assemble into complex micellar structures with mutually orthogonally oriented anisotropies at different length scales (Chapter 4). Finally, we present the results from the first scattering experiments on a set of fuel-soluble, associating telechelic polymers. We observed the formation of supramolecular aggregates in dilute (≤0.5wt%) solutions of telechelic polymers and determined that the choice of solvent has a significant effect on the strength of association and the size of the supramolecules (Chapter 5). A method was developed for the direct estimation of supramolecular aggregation number from SANS data. The insight into structure-property relationships obtained from this work will enable the more targeted development of these molecular architectures for their respective applications.

Maitasunean eta errespetuan hezten: 0-3 urte bitartekoen harmonia

LABURPENA: Aurrerapen zientifikoek aurrera egin ahala, lehen haurtzarorako begirada eraldatu egin da, bereziki garapen motorraren alorrean. Haurtxoa izaki pasibo bezala ikusten zuten ikuskerek, subjektuaren konpetentzia goiztiarren aurrean begiruneko jarrerak gartu dituzte. Lan honetan kalitatezko hezkuntza eredu instituzionalizatua garatu zuen Emmi Pikler-en ekarpenak aztertuko ditugu. Eredu honek printzipio pedagogiko zehatz batzuen arabera jaioberria eta haurtxo txikia ikusteko beste era bat eta hezitzaile bezala zaintzen dugun ume bakoitzarekiko betetzen dugun rolaren eraldaketa proposatzen du. Honek hezitzailearen jardunbide tradizionala birplanteatzera bultzatzen gaitu, haurrenganako maitasuna eta begirunea oinarritzat duen hezkuntza sistema bat eraiki ahal izateko.