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991.
Genome physical mapping from large-insert clones by fingerprint analysis with capillary electrophoresis: a robust physical map of Penicillium chrysogenum 总被引:2,自引:0,他引:2 下载免费PDF全文
Xu Z van den Berg MA Scheuring C Covaleda L Lu H Santos FA Uhm T Lee MK Wu C Liu S Zhang HB 《Nucleic acids research》2005,33(5):e50
Physical mapping with large-insert clones is becoming an active area of genomics research, and capillary electrophoresis (CE) promises to revolutionize the physical mapping technology. Here, we demonstrate the utility of the CE technology for genome physical mapping with large-insert clones by constructing a robust, binary bacterial artificial chromosome (BIBAC)-based physical map of Penicillium chrysogenum. We fingerprinted 23.1× coverage BIBAC clones with five restriction enzymes and the SNaPshot kit containing four fluorescent-ddNTPs using the CE technology, and explored various strategies to construct quality physical maps. It was shown that the fingerprints labeled with one or two colors, resulting in 40–70 bands per clone, were assembled into much better quality maps than those labeled with three or four colors. The selection of fingerprinting enzymes was crucial to quality map construction. From the dataset labeled with ddTTP–dROX, we assembled a physical map for P.chrysogenum, with 2–3 contigs per chromosome and anchored the map to its chromosomes. This map represents the first physical map constructed using the CE technology, thus providing not only a platform for genomic studies of the penicillin-producing species, but also strategies for efficient use of the CE technology for genome physical mapping of plants, animals and microbes. 相似文献
992.
THE polysaccharides obtained from some basidiomycetes, Pleurotus ostreatus (Jacq. ex Fr.) Quel.1, Flammulina velutipes (Curt, ex Fr.) Sing.2, Lentinus edodes (Berk.) Sing.2–4, Polia cocos, Wolf (Bukuryo)5 and Schizophyllum commune Fr.7, strongly inhibit the growth of sarcoma 180 and their anti-tumour activity is host-mediated8,9. 相似文献
993.
Alexey V. Morozov Tatiana M. Astakhova David G. Garbuz George S. Krasnov Natalia V. Bobkova Olga G. Zatsepina Vadim L. Karpov Michail B. Evgen’ev 《Cell stress & chaperones》2017,22(5):687-697
The heat shock protein 70 (Hsp70, human HSPA1A) plays indispensable roles in cellular stress responses and protein quality control (PQC). In the framework of PQC, it cooperates with the ubiquitin-proteasome system (UPS) to clear damaged and dysfunctional proteins in the cell. Moreover, Hsp70 itself is rapidly degraded following the recovery from stress. It was demonstrated that its fast turnover is mediated via ubiquitination and subsequent degradation by the 26S proteasome. At the same time, the effect of Hsp70 on the functional state of proteasomes has been insufficiently investigated. Here, we characterized the direct effect of recombinant Hsp70 on the activity of 20S and 26S proteasomes and studied Hsp70 degradation by the 20S proteasome in vitro. We have shown that the activity of purified 20S proteasomes is decreased following incubation with recombinant human Hsp70. On the other hand, high concentrations of Hsp70 activated 26S proteasomes. Finally, we obtained evidence that in addition to previously reported ubiquitin-dependent degradation, Hsp70 could be cleaved independent of ubiquitination by the 20S proteasome. The results obtained reveal novel aspects of the interplay between Hsp70 and proteasomes. 相似文献
994.
Rendal Vázquez ME Díaz Román TM Rodríguez Cabarcos M Zavanella Botta C Domenech García N González Cuesta M Sánchez Dopico MJ Pértega Díaz S Andión Núñez C 《Cell and tissue banking》2008,9(2):101-107
To analyse the influence of cold ischemic time (CIT) (2–24 h) and of cryopreservation (liquid phase) on the viability of the
valvular fibroblasts and in the presence of apoptosis. Cardiac valves from 10 pigs were evaluated by anatomo-pathological
study of the wall, muscle and leaflet. At the same time, the presence of cellular death due to apoptosis was investigated
in two ways; directly on tissue by Apodetec system and by two-colour flow cytometry assay analyzing a suspension of fibroblast
from valve leaflets using Anexina V and propidium iodure (PI). We established three groups of samples to compare different
experimental conditions: 2 h of ischemia (group 1), 24 h of ischemia (group 2), and a programme of cryopreservation (−1°C/min)
after 2 h of ischemia, followed by storage in liquid nitrogen during a week and thawing was performed (group 3). The analysis
of viabilities showed slight differences between all three groups. The results indicated CIT of 24 h undergoing more structural
affectation than CIT of 2 h. Flow cytometry analysis did not show important differences between groups; however cryopreserved
samples (group 3) slightly less viability and a higher percentage of death by apoptosis than group 1 and 2 using flow cytometry.
Apoptosis was confirmed on tissue from all valves but mainly in samples of group 2 and group 3. In summary, the viability
of the valves in the case of ischemic times of 2 h, 24 h or after cryopreservation/thawing differs slightly. The death of
the cells is mainly mediated by necrosis and not by apoptosis. 相似文献
995.
Lorena Rela Sung Min Yang Lidia Szczupak 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》2009,195(2):139-150
The NS neurons are nonspiking cells, present as pairs in each midbody ganglion of the leech nervous system, which display
a very extensive arborization. They were shown to regulate the coactivation of motoneurons. Here we have investigated the
electrophysiological properties of these neurons under the hypothesis that transmission along the extensive neurites requires
the aid of voltage-dependent conductances. The results indicate that NS neurons respond to electrical stimulation with a spike-like
event, which was not an all-or-none but rather a graded phenomenon that depended on the intensity and duration of the electrical
stimulus. The spike-like response was activated at a membrane potential of approximately −50 mV; its amplitude was a logarithmic
function of the extracellular Ca2+ concentration and was unaffected by a broad range of changes in the extracellular Na+ concentration; intracellular application of tetraethylammonium (TEA) caused a large increase in its amplitude and duration.
These data indicate that NS neurons bear voltage-dependent low-threshold Ca2+ and TEA-sensitive K+ conductances that could contribute to shaping synaptic signals, or transmission along the extensive neuritic tree. 相似文献
996.
997.
998.
Németh ZH Deitch EA Lu Q Szabó C Haskó G 《American journal of physiology. Cell physiology》2002,283(2):C396-C403
Na+/H+exchanger (NHE) activation has been documented to contribute toendothelial cell injury caused by inflammatory states. However, therole of NHEs in regulation of the endothelial cell inflammatoryresponse has not been investigated. The present study tested thehypothesis that NHEs contribute to endothelial cell inflammationinduced by endotoxin or interleukin (IL)-1. NHE inhibition usingamiloride, 5-(N-ethyl-N-isopropyl)-amiloride, and5-(N-methyl-N-isobutyl)amiloride as well as thenon-amiloride NHE inhibitors cimetidine, clonidine, and harmalinesuppressed endotoxin-induced IL-8 and monocyte chemoattractant protein(MCP)-1 production by human umbilical endothelial vein cells (HUVECs). The suppressive effect of amiloride on endotoxin-induced IL-8 production was associated with a decreased accumulation of IL-8 mRNA.NHE inhibitors suppressed both inhibitory (I)B degradation andnuclear factor (NF)-B DNA binding, suggesting that a decrease inactivation of the IB-NF-B system contributed to the suppression of HUVEC inflammatory response by NHE blockade. NHE inhibition decreased also the IL-1-induced HUVEC inflammatory response, becauseamiloride suppressed IL-1-induced E-selectin expression on HUVECs.These results demonstrate that maximal activation of the HUVECinflammatory response requires a functional NHE. 相似文献
999.
Thierry Rème Dirk Hose John De Vos Aurélien Vassal Pierre-Olivier Poulain Véronique Pantesco Hartmut Goldschmidt Bernard Klein 《BMC bioinformatics》2008,9(1):16
Background
The huge amount of data generated by DNA chips is a powerful basis to classify various pathologies. However, constant evolution of microarray technology makes it difficult to mix data from different chip types for class prediction of limited sample populations. Affymetrix® technology provides both a quantitative fluorescence signal and a decision (detection call: absent or present) based on signed-rank algorithms applied to several hybridization repeats of each gene, with a per-chip normalization. We developed a new prediction method for class belonging based on the detection call only from recent Affymetrix chip type. Biological data were obtained by hybridization on U133A, U133B and U133Plus 2.0 microarrays of purified normal B cells and cells from three independent groups of multiple myeloma (MM) patients.Results
After a call-based data reduction step to filter out non class-discriminative probe sets, the gene list obtained was reduced to a predictor with correction for multiple testing by iterative deletion of probe sets that sequentially improve inter-class comparisons and their significance. The error rate of the method was determined using leave-one-out and 5-fold cross-validation. It was successfully applied to (i) determine a sex predictor with the normal donor group classifying gender with no error in all patient groups except for male MM samples with a Y chromosome deletion, (ii) predict the immunoglobulin light and heavy chains expressed by the malignant myeloma clones of the validation group and (iii) predict sex, light and heavy chain nature for every new patient. Finally, this method was shown powerful when compared to the popular classification method Prediction Analysis of Microarray (PAM).Conclusion
This normalization-free method is routinely used for quality control and correction of collection errors in patient reports to clinicians. It can be easily extended to multiple class prediction suitable with clinical groups, and looks particularly promising through international cooperative projects like the "Microarray Quality Control project of US FDA" MAQC as a predictive classifier for diagnostic, prognostic and response to treatment. Finally, it can be used as a powerful tool to mine published data generated on Affymetrix systems and more generally classify samples with binary feature values.1000.