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31.
Constance Oliver 《In vitro cellular & developmental biology. Plant》1980,16(4):297-305
Summary Methods have been developed for isolating and maintaining differentiated rat exorbital lacrimal, parotid, and pancreatic acinar
cells for up to 1 month in culture. The dissociated cells retained their differentiated morphology when cultured as suspension
cultures at 35°C with the appropriate secretagogue (exorbital lacrimal, 10−6
M carbamyl choline; pancreas 10−5
M carbamyl choline; parotid, 10−6
M isoproterenol). Under these conditions the cells remained viable and differentiated for up to 4 weeks in culture and continued
to incorporate3H-leucine at rates similar to those of freshly isolated cells. If secretagogue was omitted from the medium, the cells rapidly
degenerated. These results indicate that differentiated from the medium, the cells rapidly degenerated. These results indicate
that differentiated exocrine gland acinar cells may be maintained in vitro and utilized as a model system for the study of
secretory processes. 相似文献
32.
The marine opisthobranch molluscAplysia punctata was cultured at the Laboratoire de Biologie Marine in Concarneau, France.A. punctata veligers settled and underwent metamorphosis on the algaLomentaria articulata, but not onUlva spp., Palmaria marina, Laminaria spp. andFucus spp.Research supported by grants from The Arts Foundation and the Lerner Fund for Marine Research of the American Museum of Natural History. We wish to thank Director Yves Legal, College de France for his support and cooperation. 相似文献
33.
34.
Synthesis and turnover of the regularly arranged surface protein of Acinetobacter sp. relative to the other components of the cell envelope. 总被引:6,自引:5,他引:1 下载免费PDF全文
The formation of the components of the cell envelope of Acinetobacter sp. 199A was investigated by measuring the incorporation of [3H]leucine into protein, [14C]galactose into lipopolysaccharide, 32P into phospholipid, and [3H]diaminopimelic acid into peptidoglycan. Whereas the lipopolysaccharide and intrinsic protein of the outer membrane were stable, some of the regularly arranged surface protein, the alpha-protein, was lost into the growth medium. Only newly synthesized alpha-protein was lost. The peptidoglycan of the murein layer was also labile. Selective inhibition of the formation of individual components of the cell envelope with penicillin, chloramphenicol, and bacitracin showed that incorporation of protein into the outer membrane required the simultaneous formation of complete lipopolysaccharide. The converse was not true: protein synthesis was not required for lipopolysaccharide incorporation. Formation of the outer membrane and the murein layer proceeded independently. 相似文献
35.
Aaron Mendez-Bermudez Liudmyla Lototska Melanie Pousse Florent Tessier Oliver Croce Chrysa
M Latrick Veronica Cherdyntseva Joe Nassour Jiang Xiaohua Yiming Lu Corinne Abbadie Sarantis Gagos Jing Ye Eric Gilson 《Nucleic acids research》2022,50(13):7493
Cellular senescence triggers various types of heterochromatin remodeling that contribute to aging. However, the age-related mechanisms that lead to these epigenetic alterations remain elusive. Here, we asked how two key aging hallmarks, telomere shortening and constitutive heterochromatin loss, are mechanistically connected during senescence. We show that, at the onset of senescence, pericentromeric heterochromatin is specifically dismantled consisting of chromatin decondensation, accumulation of DNA breakages, illegitimate recombination and loss of DNA. This process is caused by telomere shortening or genotoxic stress by a sequence of events starting from TP53-dependent downregulation of the telomere protective protein TRF2. The resulting loss of TRF2 at pericentromeres triggers DNA breaks activating ATM, which in turn leads to heterochromatin decondensation by releasing KAP1 and Lamin B1, recombination and satellite DNA excision found in the cytosol associated with cGAS. This TP53–TRF2 axis activates the interferon response and the formation of chromosome rearrangements when the cells escape the senescent growth arrest. Overall, these results reveal the role of TP53 as pericentromeric disassembler and define the basic principles of how a TP53-dependent senescence inducer hierarchically leads to selective pericentromeric dismantling through the downregulation of TRF2. 相似文献
36.
Tapio Schneider Oliver R. A. Dunbar Jinlong Wu Lucas Bttcher Dmitry Burov Alfredo Garbuno-Inigo Gregory L. Wagner Sen Pei Chiara Daraio Raffaele Ferrari Jeffrey Shaman 《PLoS computational biology》2022,18(6)
Testing, contact tracing, and isolation (TTI) is an epidemic management and control approach that is difficult to implement at scale because it relies on manual tracing of contacts. Exposure notification apps have been developed to digitally scale up TTI by harnessing contact data obtained from mobile devices; however, exposure notification apps provide users only with limited binary information when they have been directly exposed to a known infection source. Here we demonstrate a scalable improvement to TTI and exposure notification apps that uses data assimilation (DA) on a contact network. Network DA exploits diverse sources of health data together with the proximity data from mobile devices that exposure notification apps rely upon. It provides users with continuously assessed individual risks of exposure and infection, which can form the basis for targeting individual contact interventions. Simulations of the early COVID-19 epidemic in New York City are used to establish proof-of-concept. In the simulations, network DA identifies up to a factor 2 more infections than contact tracing when both harness the same contact data and diagnostic test data. This remains true even when only a relatively small fraction of the population uses network DA. When a sufficiently large fraction of the population (≳ 75%) uses network DA and complies with individual contact interventions, targeting contact interventions with network DA reduces deaths by up to a factor 4 relative to TTI. Network DA can be implemented by expanding the computational backend of existing exposure notification apps, thus greatly enhancing their capabilities. Implemented at scale, it has the potential to precisely and effectively control future epidemics while minimizing economic disruption. 相似文献
37.
Hamblett KJ Kegley BB Hamlin DK Chyan MK Hyre DE Press OW Wilbur DS Stayton PS 《Bioconjugate chemistry》2002,13(3):588-598
Pretargeted radioimmunotherapy specifically targets radiation to tumors using antibody-streptavidin conjugates followed by radiolabeled biotin. A potential barrier to this cancer therapy is the presence of endogenous biotin in serum, which can block the biotin-binding sites of the antibody-streptavidin conjugate before the administration of radiolabeled biotin. Serum-derived biotin can also be problematic in clinical diagnostic applications. Due to the extremely slow dissociation of the biotin-streptavidin complex, this endogenous biotin can irreversibly block the biotin-binding sites of streptavidin and reduce therapeutic efficacy, as well as reduce sensitivity in diagnostic assays. We tested a streptavidin mutant (SAv-Y43A), which has a 67-fold lower affinity for biotin than wild type streptavidin, and three bivalent bis-biotin constructs as replacements for wild-type streptavidin and biotin used in pretargeting and clinical diagnostics. Biotin dimers were engineered with certain parameters including water solubility, biotinidase resistance, and linker lengths long enough to span the distance between two biotin-binding sites of streptavidin. The bivalent biotins were compared to biotin in exchange, retention, and off-rate assays. The faster off-rate of SAv-Y43A allowed efficient exchange of prebound biotin by the biotin dimers. In fluorescent competition experiments, the biotin dimer ligands displayed high avidity binding and essentially irreversible retention with SAv-Y43A. The off-rate of a biotinidase-stabilized biotin dimer from SAv-Y43A was 4.36 x 10(-)(6) s(-)(1), over 640 times slower compared to biotin. These findings strongly suggest that employing a mutant streptavidin in concert with a bivalent biotin can mitigate the deleterious impact of endogenous biotin, by allowing exchange of bound biotin and retention of the biotin dimer carriers. 相似文献
38.
Clades that predate the origin of biomes that they inhabit provide unique opportunities to examine both when major environmental transitions occurred, and how lineages adapted to these changes. The isolated island continent Australia has undergone a profound environmental transition through the Miocene, from relatively mesic to predominantly arid; however, we have much to learn about both the timing of this change, and how organisms may have responded to it. The family Carphodactylidae is an ancient Gondwanan group of geckos that occurs across all major Australian biomes. A multilocus (ND2, Rag-1, c-mos) phylogenetic and dating analysis of the most ecologically diverse clade within this group, the genus Nephrurus (sensuBauer, 1990) reveals that two of three morphological taxa historically recognized (the 'spiny knob-tails' and 'Underwoodisaurus') are relatively species depauperate, pleisomorphic basal grades that diversified through the late Oligocene and early Miocene, and are now absent from most of the arid biome. Based on their deep divergence and morphological distinctiveness we recognize two lineages (milii and sphyrurus) as monotypic genera, the later of which is named herein (Uvidicolus nov. gen). In contrast, a third morphological group, the 'smooth knob-tails,' is a monophyletic group of five exclusively arid zone burrowing species that has radiated relatively recently (mid-Miocene). Our phylogeny indicates that successful colonization of this novel and challenging biome by Nephrurus correlates with an initial shift to terrestriality and adaptation to at least seasonally arid conditions around the early Miocene, and the eventual evolution and subsequent mid-Miocene radiation of a lineage specialized for burrowing. 相似文献
39.
Vielhauer O Zakhartsev M Horn T Takors R Reuss M 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2011,879(32):3859-3870
In the field of metabolomics, GC-MS has rather established itself as a tool for semi-quantitative strategies like metabolic fingerprinting or metabolic profiling. Absolute quantification of intra- or extracellular metabolites is nowadays mostly accomplished by application of diverse LC-MS techniques. Only few groups have so far adopted GC-MS technology for this exceptionally challenging task. Besides numerous and deeply investigated problems related to sample generation, the pronounced matrix effects in biological samples have led to the almost mandatory application of isotope dilution mass spectrometry (IDMS) for the accurate determination of absolute metabolite concentrations. Nevertheless, access to stable isotope labeled internal standards (ILIS), which are in many cases commercially unavailable, is quite laborious and very expensive. Here we present an improved and simplified gas chromatography-isotope dilution mass spectrometry (GC-IDMS) protocol for the absolute determination of intra- and extracellular metabolite levels. Commercially available (13)C-labeled algal cells were used as a convenient source for the preparation of internal standards. Advantages as well as limitations of the described method are discussed. 相似文献
40.
Synaptic destabilization by neuronal Nogo-A 总被引:1,自引:0,他引:1
Aloy EM Weinmann O Pot C Kasper H Dodd DA Rülicke T Rossi F Schwab ME 《Brain Cell Biology》2006,35(2-3):137-157
Formation and maintenance of a neuronal network is based on a balance between plasticity and stability of synaptic connections.
Several molecules have been found to regulate the maintenance of excitatory synapses but nothing is known about the molecular
mechanisms involved in synaptic stabilization versus disassembly at inhibitory synapses. Here, we demonstrate that Nogo-A,
which is well known to be present in myelin and inhibit growth in the adult CNS, is present in inhibitory presynaptic terminals
in cerebellar Purkinje cells at the time of Purkinje cell-Deep Cerebellar Nuclei (DCN) inhibitory synapse formation and is
then downregulated during synapse maturation. We addressed the role of neuronal Nogo-A in synapse maturation by generating
several mouse lines overexpressing Nogo-A, starting at postnatal ages and throughout adult life, specifically in cerebellar
Purkinje cells and their terminals. The overexpression of Nogo-A induced a progressive disassembly, retraction and loss of
the inhibitory Purkinje cell terminals. This led to deficits in motor learning and coordination in the transgenic mice. Prior
to synapse disassembly, the overexpression of neuronal Nogo-A led to the downregulation of the synaptic scaffold proteins
spectrin, spectrin-E and β-catenin in the postsynaptic neurons. Our data suggest that neuronal Nogo-A might play a role in
the maintenance of inhibitory synapses by modulating the expression of synaptic anchoring molecules.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献