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101.
Mona El Khatib Przemyslaw Bozko Vindhya Palagani Nisar P. Malek Ludwig Wilkens Ruben R. Plentz 《PloS one》2013,8(10)
Cholangiocacinoma (CC) is a cancer disease with rising incidence. Notch signaling has been shown to be deregulated in many cancers. However, the role of this signaling pathway in the carcinogenesis of CC is still not fully explored. In this study, we investigated the effects of Notch inhibition by γ-secretase inhibitor IX (GSI IX) in cultured human CC cell lines and we established a transgenic mouse model with liver specific expression of the intracellular domain of Notch (Notch-ICD) and inactivation of tumor suppressor p53. GSI IX treatment effectively impaired cell proliferation, migration, invasion, epithelial to mesenchymal transition and growth of softagar colonies. In vivo overexpression of Notch-ICD together with an inactivation of p53 significantly increased tumor burden and showed CC characteristics. Conclusion: Our study highlights the importance of Notch signaling in the tumorigenesis of CC and demonstrates that additional inactivation of p53 in vivo. 相似文献
102.
Meike V. F. Schlupp Julia Martynczuk Michel Prestat Ludwig J. Gauckler 《Liver Transplantation》2013,3(3):375-385
Microstructures of yttria stabilized zirconia thin films deposited by aerosol assisted chemical vapor deposition (AA‐CVD) are correlated with the thermal decomposition behavior of the corresponding metal precursors, zirconium and yttrium 2,4‐pentanedionate. Process conditions of AA‐CVD are investigated with the aim of producing dense and compact YSZ thin films for applications as gas‐tight electrolyte. Based on systematic cross sectional scanning transmission electron microscopy (STEM) investigations and conductivity measurements, the development of percolating nanoporosity is observed in samples prepared at temperatures between 350 °C and 600 °C at standard solution throughput. Compact columnar thin films with bulk conductivity are obtained at 600 °C by reducing the metal content of the precursor solution and at 450 °C by reducing the solution throughput. 相似文献
103.
Giovanni Monaco Elke Decrock Koen Nuyts Larry E. Wagner II Tomas Luyten Sergei V. Strelkov Ludwig Missiaen Wim M. De Borggraeve Luc Leybaert David I. Yule Humbert De Smedt Jan B. Parys Geert Bultynck 《PloS one》2013,8(8)
The anti-apoptotic Bcl-2 protein is the founding member and namesake of the Bcl-2-protein family. It has recently been demonstrated that Bcl-2, apart from its anti-apoptotic role at mitochondrial membranes, can also directly interact with the inositol 1,4,5-trisphosphate receptor (IP3R), the primary Ca2+-release channel in the endoplasmic reticulum (ER). Bcl-2 can thereby reduce pro-apoptotic IP3R-mediated Ca2+ release from the ER. Moreover, the Bcl-2 homology domain 4 (Bcl-2-BH4) has been identified as essential and sufficient for this IP3R-mediated anti-apoptotic activity. In the present study, we investigated whether the reported inhibitory effect of a Bcl-2-BH4 peptide on the IP 3R1 was related to the distinctive α-helical conformation of the BH4 domain peptide. We therefore designed a peptide with two glycine “hinges” replacing residues I14 and V15, of the wild-type Bcl-2-BH4 domain (Bcl-2-BH4-IV/GG). By comparing the structural and functional properties of the Bcl-2-BH4-IV/GG peptide with its native counterpart, we found that the variant contained reduced α-helicity, neither bound nor inhibited the IP 3R1 channel, and in turn lost its anti-apoptotic effect. Similar results were obtained with other substitutions in Bcl-2-BH4 that destabilized the α-helix with concomitant loss of IP3R inhibition. These results provide new insights for the further development of Bcl-2-BH4-derived peptides as specific inhibitors of the IP3R with significant pharmacological implications. 相似文献
104.
Roberto Montanhini Neto Thayzi O. Zeni Sandra Ludwig Aline Horodesky Marcus V.F. Girotto Gisele G. Castilho-Westphal 《Invertebrate reproduction & development.》2013,57(3):208-218
The production of oysters in Guaratuba Bay, in the state of Paraná, Brazil, is still a mixed activity of mariculture and extractivism. The sustainable development of this production requires the monitoring of environmental, genetic, reproductive, and zootechnical variables. This study evaluated the importance of these variables on Crassostrea oyster production. Data were obtained between September 2009 and February 2011 from six evaluations: water quality, oyster larvae in plankton, capture of oyster spat by artificial collectors, molecular identification of collected spat and larvae, continuous evaluation of the reproductive maturity stage of adult oysters, and zootechnical performance achieved by experimental production. Temperature, dissolved oxygen, and water transparency were the environmental factors that had the most influence on the reproductive cycle (e.g. gonadal development and spat capture) and on the growth of the oyster. The highest rates for spat uptake were recorded in the summer, coinciding with the peak of sexual maturity of adults. The growth of the oysters, in terms of the weight of the meat, was related to the gonadal stage. 相似文献
105.
Santeri Kiviluoto Tomas Luyten Lars Schneider Dmitrij Lisak Diego Rojas-Rivera Kirsten Welkenhuyzen Ludwig Missaen Humbert De Smedt Jan B. Parys Claudio Hetz Axel Methner Geert Bultynck 《Cell calcium》2013
Bax Inhibitor-1 (BI-1) is an evolutionarily conserved six-transmembrane domain endoplasmic reticulum (ER)-localized protein that protects against ER stress-induced apoptotic cell death. This function is closely connected to its ability to lower steady-state ER Ca2+ levels. Recently, we elucidated BI-1's Ca2+-channel pore in the C-terminal part of the protein and identified the critical amino acids of its pore. Based on these insights, a Ca2+-channel pore-dead mutant BI-1 (BI-1D213R) was developed. We determined whether BI-1 behaves as a bona fide H+/Ca2+ antiporter or as an ER Ca2+-leak channel by investigating the effect of pH on unidirectional Ca2+-efflux rates. At pH 6.8, wild-type BI-1 expression in BI-1−/− cells increased the ER Ca2+-leak rate, correlating with its localization in the ER compartment. In contrast, BI-1D231R expression in BI-1−/−, despite its ER localization, did not increase the ER Ca2+-leak rate. However, at pH < 6.8, the BI-1-mediated ER Ca2+ leak was blocked. Finally, a peptide representing the Ca2+-channel pore of BI-1 promoting Ca2+ flux from the ER was used. Lowering the pH from 6.8 to 6.0 completely abolished the ability of the BI-1 peptide to mediate Ca2+ flux from the ER. We propose that this pH dependence is due to two aspartic acid residues critical for the function of the Ca2+-channel pore and located in the ER membrane-dipping domain, which facilitates the protonation of these residues. 相似文献
106.
Carmen F. Ludwig Florian Ullrich Lilia Leisle Tobias Stauber Thomas J. Jentsch 《The Journal of biological chemistry》2013,288(40):28611-28619
CLC anion transporters form dimers that function either as Cl− channels or as electrogenic Cl−/H+ exchangers. CLC channels display two different types of “gates,” “protopore” gates that open and close the two pores of a CLC dimer independently of each other and common gates that act on both pores simultaneously. ClC-7/Ostm1 is a lysosomal 2Cl−/1H+ exchanger that is slowly activated by depolarization. This gating process is drastically accelerated by many CLCN7 mutations underlying human osteopetrosis. Making use of some of these mutants, we now investigate whether slow voltage activation of plasma membrane-targeted ClC-7/Ostm1 involves protopore or common gates. Voltage activation of wild-type ClC-7 subunits was accelerated by co-expressing an excess of ClC-7 subunits carrying an accelerating mutation together with a point mutation rendering these subunits transport-deficient. Conversely, voltage activation of a fast ClC-7 mutant could be slowed by co-expressing an excess of a transport-deficient mutant. These effects did not depend on whether the accelerating mutation localized to the transmembrane part or to cytoplasmic cystathionine-β-synthase (CBS) domains of ClC-7. Combining accelerating mutations in the same subunit did not speed up gating further. No currents were observed when ClC-7 was truncated after the last intramembrane helix. Currents and slow gating were restored when the C terminus was co-expressed by itself or fused to the C terminus of the β-subunit Ostm1. We conclude that common gating underlies the slow voltage activation of ClC-7. It depends on the CBS domain-containing C terminus that does not require covalent binding to the membrane domain of ClC-7. 相似文献
107.
The Yangtze sturgeon (Acipenser dabryanus) is an endemic species in China. Using 454 sequencing, eight polymorphic tri‐, tetra‐, penta‐, and hexanucleotide microsatellite loci were isolated in this study. The raw sequence data from a one‐eighth run of 454 sequencings were 38.0 Mbp containing 94 222 reads/sequences. Of 80 microsatellite loci, only eight loci were polymorphic in a population of 30 individuals. The number of alleles per locus ranged from 4 to 14 (mean 7.62), and the observed heterozygosities varied between 0.46 and 0.88 (mean 0.74). Cross amplification was tested in congeneric species Acipenser sturio and Acipenser sinensis. These new microsatellite markers will be useful for further studies on genetic variation, parentage analysis, and conservation management for this critically endangered species. 相似文献
108.
Sonja C. Ludwig Aly McCluskie Paula Keane Catherine Barlow Richard M. Francksen Damian Bubb 《Bird Study》2013,60(4):431-443
Capsule: Diversionary feeding reduced Hen Harrier Circus cyaneus nestlings’ natural food intake by half. Red Grouse Lagopus lagopus scotica chicks constituted 0–4% of all nestling food items. Annually, this reduced annual grouse chick production by 0–6%.Aim: To quantify proportions of diversionary and natural food (including grouse) delivered to Hen Harrier nestlings in relation to brood size, male status and natural prey abundance.Methods: We recorded diversionary food provisioned to 25 Hen Harrier broods (2008–15) and studied the diet of 15 broods using observations from hides, nest cameras and regurgitated pellet analysis. Variation in nestling diet was analysed using compositional analysis.Results: Hen Harriers took 76% of diversionary food provided. Depending on assessment method, average nestling diet was 44–53% diversionary food, 39–55% natural prey (including 24–45% passerines, 4–15% small mammals, 0–4% grouse chicks) and 0–9% unknown items. The amount of diversionary food consumed was not influenced by male status, brood size or natural prey abundance. The number of Red Grouse chicks delivered annually was 34–100% lower than expected under unfed conditions, however, the confidence intervals associated with these estimates were large.Conclusion: Diversionary food influenced Hen Harrier nestling diet and reduced the number of Red Grouse chicks taken relative to modelled predictions. It may help reduce conflict between Hen Harrier conservation and Red Grouse shooting, but only if overall grouse productivity is thereby maintained or increased. 相似文献
109.
Simona Maria Monti Giuseppina De Simone Nina A. Dathan Martha Ludwig Daniela Vullo Andrea Scozzafava Clemente Capasso Claudiu T. Supuran 《Bioorganic & medicinal chemistry letters》2013,23(6):1626-1630
Several β-carbonic anhydrases (CAs, EC 4.2.1.1) are present in all land plants examined thus far. Here we report the first detailed biochemical characterization of one such isoform, FbiCA 1, from the C4 plant Flaveria bidentis, which was cloned, purified and characterized as recombinant protein. FbiCA 1 has an interesting CO2 hydrase catalytic activity (kcat of 1.2 × 105 and kcat/Km of 7.5 × 106 M?1 × s?1) and was moderately inhibited by most simple/complex inorganic anions. Potent FbiCA 1 inhibitors were also detected, such as trithiocarbonate, diethyldithiocarbamate, sulfamide, sulfamic acid, phenylboronic acid and phenylarsonic acid (KIs in the range of 4–60 μM). Such inhibitors may be used as tools to better understand the role of various β-CA isoforms in photosynthesis. 相似文献
110.
Alexandra Wasmuth Christina Ludwig Henning D. Mootz 《Bioorganic & medicinal chemistry》2013,21(12):3495-3503
Protein trans-splicing by split inteins holds great potential for the chemical modification and semisynthesis of proteins. However, the structural requirements of the extein sequences immediately flanking the intein are only poorly understood. This knowledge is of particular importance for protein labeling, when synthetic moieties are to be attached to the protein of interest as seamlessly as possible. Using the semisynthetic Ssp DnaB intein both in form of its wild-type sequence and its evolved M86 mutant, we systematically varied the sequence upstream of the short synthetic IntN fragment using both proteinogenic amino acids and unnatural building blocks. We could show for the wild-type variant that the native N-extein sequence could be reduced to the glycine residue at the (?1) position directly flanking the intein without significant loss of activity. The glycine at this position is strongly preferred over building blocks containing a phenyl group or extended alkyl chain adjacent to the scissile amide bond of the N-terminal splice junction. Despite their negative effects on the splicing yields, these unnatural substrates were well processed in the N–S acyl shift to form the respective thioesters and did not result in an increased decoupling of the asparagine cyclization step at the C-terminal splicing junction. Therefore, the transesterification step appeared to be the bottleneck of the protein splicing pathway. The fluorophore 7-hydroxycoumarinyl-4-acetic acid as a minimal N-extein was efficiently ligated to the model protein, in particular with the M86 mutant, probably because of its higher resemblance to glycine with an aliphatic c-α carbon atom at the (?1) position. This finding indicates a way for the virtually traceless labeling of proteins without inserting extra flanking residues. Due to its overall higher activity, the M86 mutant appears most promising for many protein labeling and chemical modification schemes using the split intein approach. 相似文献