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141.
Attaining metabolic and isotopic balanced growth is one critical condition for physiological studies using isotope-labeled tracers, but is very difficult to obtain in batch culture due to the extensive metabolite exchange with the surrounding medium and related physiological changes. In the present study, we investigated metabolic and isotopic behavior of CHO cells in differently designed media. We observed that the assumption of balanced cell growth cannot be justified in batch culture of CHO cells directly using conventional, commercially available media. By systematically redesigning media composition and characterizing metabolic steady state based on mass balances and measurement of labeling dynamics, we achieved balanced cell growth for the main cellular substrates in CHO cells. This was done in a step-by-step analysis of growth and primary metabolism of CHO cells with the use of [U-13C]glucose feeding and adjusting concentrations of amino acids in the growth medium. The optimized media obtained at the end of the study provide balanced growth and isotopic steady state or at least asymptotic steady state. As a result, we established a platform to conduct isotope-based physiological studies of mammalian systems more reliably and therefore well suited for later use in metabolic profiling of mammalian systems such as 13C-labeled metabolic flux analysis. 相似文献
142.
Biosorption of dyes using dead macro fungi: effect of dye structure, ionic strength and pH 总被引:10,自引:0,他引:10
Biosorbents prepared from dead macro fungi, namely Fomes fomentarius and Phellinus igniarius, were applied for the uptake of Methylene Blue (MB) and Rhodamine B (RB). Equilibrium isotherm data could be well described by the Langmuir and Freundlich models. Methylene Blue was found to be more adsorbable than Rhodamine B. Langmuir monolayer coverage was determined as 204.38-232.73 mg/g and 25.12-36.82 mg/g for MB and RB, respectively. Molecular structure and ionic radius of dyes were found to be responsible for differences in their uptakes. Results showed that sorption of MB increased while that of RB decreased as pH of respective dye solutions changed from 3 to 11. An increase in ionic strength also exhibited an adverse effect on dye sorption capacity. Ionic strength and pH affected the sorption of MB more as compared to the sorption of RB. The presence of carboxylic (-ve) and amino (+ve) groups in RB could explain the lower sorption of RB compared to MB. 相似文献
143.
Cédric Berney Andreea Ciuprina Sara Bender Juliet Brodie Virginia Edgcomb Eunsoo Kim Jeena Rajan Laura Wegener Parfrey Sina Adl Stéphane Audic David Bass David A. Caron Guy Cochrane Lucas Czech Micah Dunthorn Stefan Geisen Frank Oliver Glöckner Frédéric Mahé Christian Quast Jonathan Z. Kaye Alastair G. B. Simpson Alexandros Stamatakis Javier del Campo Pelin Yilmaz Colomban de Vargas 《The Journal of eukaryotic microbiology》2017,64(3):407-411
Universal taxonomic frameworks have been critical tools to structure the fields of botany, zoology, mycology, and bacteriology as well as their large research communities. Animals, plants, and fungi have relatively solid, stable morpho‐taxonomies built over the last three centuries, while bacteria have been classified for the last three decades under a coherent molecular taxonomic framework. By contrast, no such common language exists for microbial eukaryotes, even though environmental ‘‐omics’ surveys suggest that protists make up most of the organismal and genetic complexity of our planet's ecosystems! With the current deluge of eukaryotic meta‐omics data, we urgently need to build up a universal eukaryotic taxonomy bridging the protist ‐omics age to the fragile, centuries‐old body of classical knowledge that has effectively linked protist taxa to morphological, physiological, and ecological information. UniEuk is an open, inclusive, community‐based and expert‐driven international initiative to build a flexible, adaptive universal taxonomic framework for eukaryotes. It unites three complementary modules, EukRef, EukBank, and EukMap, which use phylogenetic markers, environmental metabarcoding surveys, and expert knowledge to inform the taxonomic framework. The UniEuk taxonomy is directly implemented in the European Nucleotide Archive at EMBL‐EBI, ensuring its broad use and long‐term preservation as a reference taxonomy for eukaryotes. 相似文献
144.
Mandenius CF Andersson TB Alves PM Batzl-Hartmann C Björquist P Carrondo MJ Chesne C Coecke S Edsbagge J Fredriksson JM Gerlach JC Heinzle E Ingelman-Sundberg M Johansson I Küppers-Munther B Müller-Vieira U Noor F Zeilinger K 《Alternatives to laboratory animals : ATLA》2011,39(2):147-171
Drug-induced liver injury is a common reason for drug attrition in late clinical phases, and even for post-launch withdrawals. As a consequence, there is a broad consensus in the pharmaceutical industry, and within regulatory authorities, that a significant improvement of the current in vitro test methodologies for accurate assessment and prediction of such adverse effects is needed. For this purpose, appropriate in vivo-like hepatic in vitro models are necessary, in addition to novel sources of human hepatocytes. In this report, we describe recent and ongoing research toward the use of human embryonic stem cell (hESC)-derived hepatic cells, in conjunction with new and improved test methods, for evaluating drug metabolism and hepatotoxicity. Recent progress on the directed differentiation of human embryonic stem cells to the functional hepatic phenotype is reported, as well as the development and adaptation of bioreactors and toxicity assay technologies for the testing of hepatic cells. The aim of achieving a testing platform for metabolism and hepatotoxicity assessment, based on hESC-derived hepatic cells, has advanced markedly in the last 2-3 years. However, great challenges still remain, before such new test systems could be routinely used by the industry. In particular, we give an overview of results from the Vitrocellomics project (EU Framework 6) and discuss these in relation to the current state-of-the-art and the remaining difficulties, with suggestions on how to proceed before such in vitro systems can be implemented in industrial discovery and development settings and in regulatory acceptance. 相似文献
145.
Fikret G. Agircan Elmar Schiebel Balca R. Mardin 《Philosophical transactions of the Royal Society of London. Series B, Biological sciences》2014,369(1650)
The centrosome is the main microtubule (MT)-organizing centre of animal cells. It consists of two centrioles and a multi-layered proteinaceous structure that surrounds the centrioles, the so-called pericentriolar material. Centrosomes promote de novo assembly of MTs and thus play important roles in Golgi organization, cell polarity, cell motility and the organization of the mitotic spindle. To execute these functions, centrosomes have to adopt particular cellular positions. Actin and MT networks and the association of the centrosomes to the nuclear envelope define the correct positioning of the centrosomes. Another important feature of centrosomes is the centrosomal linker that connects the two centrosomes. The centrosome linker assembles in late mitosis/G1 simultaneously with centriole disengagement and is dissolved before or at the beginning of mitosis. Linker dissolution is important for mitotic spindle formation, and its cell cycle timing has profound influences on the execution of mitosis and proficiency of chromosome segregation. In this review, we will focus on the mechanisms of centrosome positioning and separation, and describe their functions and mechanisms in the light of recent findings. 相似文献
146.
Nina Weller Petra Clowes Gerhard Dobler Elmar Saathoff Inge Kroidl Nyanda Elias Ntinginya Leonard Maboko Thomas L?scher Michael Hoelscher Norbert Heinrich 《PLoS neglected tropical diseases》2014,8(7)
Background
To date, Alphavirus infections and their most prominent member, chikungunya fever, a viral disease which first became apparent in Tanzania in 1953, have been very little investigated in regions without epidemic occurrence. Few data exist on burden of disease and socio-economic and environmental covariates disposing to infection.Methods
A cross-sectional seroprevalence study was undertaken in 1,215 persons from Mbeya region, South-Western Tanzania, to determine the seroprevalence of anti-Alphavirus IgG antibodies, and to investigate associated risk factors.Results
18% of 1,215 samples were positive for Alphavirus IgG. Seropositivity was associated with participant age, low to intermediate elevation, flat terrain and with IgG positivity for Rift Valley fever, Flaviviridae, and rickettsiae of the spotted fever group. When comparing the geographical distribution of Alphavirus seropositivity to that of Rift Valley fever, it was obvious that Alphaviruses had spread more widely throughout the study area, while Rift Valley fever was concentrated along the shore of Lake Malawi.Conclusion
Alphavirus infections may contribute significantly to the febrile disease burden in the study area, and are associated with several arthropod-borne infections. Their spread seems only limited by factors affecting mosquitoes, and seems less restricted than that of Rift Valley fever. 相似文献147.
Wenxin Liu Manje Gowda Jochen C Reif Volker Hahn Arno Ruckelshausen Elmar A Weissmann Hans Peter Maurer Tobias Würschum 《BMC genomics》2014,15(1)
Background
The nature of dynamic traits with their phenotypic plasticity suggests that they are under the control of a dynamic genetic regulation. We employed a precision phenotyping platform to non-invasively assess biomass yield in a large mapping population of triticale at three developmental stages.Results
Using multiple-line cross QTL mapping we identified QTL for each of these developmental stages which explained a considerable proportion of the genotypic variance. Some QTL were identified at each developmental stage and thus contribute to biomass yield throughout the studied developmental phases. Interestingly, we also observed QTL that were only identified for one or two of the developmental stages illustrating a temporal contribution of these QTL to the trait. In addition, epistatic QTL were detected and the epistatic interaction landscape was shown to dynamically change with developmental progression.Conclusions
In summary, our results reveal the temporal dynamics of the genetic architecture underlying biomass accumulation in triticale and emphasize the need for a temporal assessment of dynamic traits.Electronic supplementary material
The online version of this article (doi:10.1186/1471-2164-15-458) contains supplementary material, which is available to authorized users. 相似文献148.
Wilhelm M. Malloni Silvia De Sanctis Ana M. Tomé Elmar W. Lang Claudia E. Munte Klaus Peter Neidig Hans Robert Kalbitzer 《Journal of biomolecular NMR》2010,47(2):101-111
Strong solvent signals lead to a disappearance of weak protein signals close to the solvent resonance frequency and to base
plane variations all over the spectrum. AUREMOL-SSA provides an automated approach for solvent artifact removal from multidimensional
NMR protein spectra. Its core algorithm is based on singular spectrum analysis (SSA) in the time domain and is combined with
an automated base plane correction in the frequency domain. The performance of the method has been tested on synthetic and
experimental spectra including two-dimensional NOESY and TOCSY spectra and a three-dimensional 1H,13C-HCCH-TOCSY spectrum. It can also be applied to frequency domain spectra since an optional inverse Fourier transformation
is included in the algorithm. 相似文献
149.
Roderick C. Slieker Matthias S. Roost Liesbeth van Iperen H. Eka D. Suchiman Elmar W. Tobi Fran?oise Carlotti Eelco J. P. de Koning P. Eline Slagboom Bastiaan T. Heijmans Susana M. Chuva de Sousa Lopes 《PLoS genetics》2015,11(10)
Remodelling the methylome is a hallmark of mammalian development and cell differentiation. However, current knowledge of DNA methylation dynamics in human tissue specification and organ development largely stems from the extrapolation of studies in vitro and animal models. Here, we report on the DNA methylation landscape using the 450k array of four human tissues (amnion, muscle, adrenal and pancreas) during the first and second trimester of gestation (9,18 and 22 weeks). We show that a tissue-specific signature, constituted by tissue-specific hypomethylated CpG sites, was already present at 9 weeks of gestation (W9). Furthermore, we report large-scale remodelling of DNA methylation from W9 to W22. Gain of DNA methylation preferentially occurred near genes involved in general developmental processes, whereas loss of DNA methylation mapped to genes with tissue-specific functions. Dynamic DNA methylation was associated with enhancers, but not promoters. Comparison of our data with external fetal adrenal, brain and liver revealed striking similarities in the trajectory of DNA methylation during fetal development. The analysis of gene expression data indicated that dynamic DNA methylation was associated with the progressive repression of developmental programs and the activation of genes involved in tissue-specific processes. The DNA methylation landscape of human fetal development provides insight into regulatory elements that guide tissue specification and lead to organ functionality. 相似文献
150.
Lynch AM Guzzie PJ Bauer D Gocke E Itoh S Jacobs A Krul CA Schepky A Tanaka N Kasper P 《Mutation research》2011,723(2):91-100
A workshop to reappraise the previous IWGT recommendations for photogenotoxicity testing [E. Gocke, L. Muller, P.J. Guzzie, S. Brendler-Schwaab, S. Bulera, C.F. Chignell, L.M. Henderson, A. Jacobs, H. Murli, R.D. Snyder, N. Tanaka, Considerations on photochemical genotoxicity: report of the International Workshop on Genotoxicity Test Procedures working group, Environ. Mol. Mutagen., 35 (2000) 173-184] was recently held as part of the 5th International Workshop on Genotoxicity Testing (IWGT) meeting in Basel, Switzerland (August 17-19, 2009). An Expert Panel was convened from regulatory, academic and industrial scientists (with several members serving on the original panel) and chaired by Dr Peter Kasper (BfArM, Germany). The aim of the workshop was to review progress made in photo(geno)toxicity testing over the past decade; a period which saw the introduction of several regulatory photosafety guidances in particular in Europe and the USA. Based on current regulatory guidelines a substantial proportion of compounds trigger the requirements for photosafety testing. Moreover, there has been growing concern within industry about the performance of the in vitro photosafety tests in the "real world" of compound development. Therefore, the expert group reviewed the status of the current regulatory guidance's and the impact these have had on compound development in the context of the various triggers for photosafety testing. In addition, the performance of photogenotoxicity assays (old and new) was discussed, particularly in view of reports of pseudophotoclastogencity. The Expert Panel finished with an assessment of the positioning of photogenotoxicity testing within a photosafety testing strategy. The most significant conclusion made by the Expert Panel was that photogenotoxicity testing should no longer be recommended as part of the standard photosafety testing strategy. In addition, progress was made on the refinement of triggers for photosafety testing. For example, there was support for harmonisation of methods to determine the Molar Extinction Coefficient (MEC) and a consensus agreement that there should be no requirement for testing of compounds with a MEC<1000Lmol(-1)cm(-1). 相似文献