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141.
Bindewald E Wendeler M Legiewicz M Bona MK Wang Y Pritt MJ Le Grice SF Shapiro BA 《RNA (New York, N.Y.)》2011,17(9):1688-1696
Selective 2'-hydroxyl acylation analyzed by primer extension (SHAPE) is a facile technique for quantitative analysis of RNA secondary structure. In general, low SHAPE signal values indicate Watson-Crick base-pairing, and high values indicate positions that are single-stranded within the RNA structure. However, the relationship of SHAPE signals to structural properties such as non-Watson-Crick base-pairing or stacking has thus far not been thoroughly investigated. Here, we present results of SHAPE experiments performed on several RNAs with published three-dimensional structures. This strategy allows us to analyze the results in terms of correlations between chemical reactivities and structural properties of the respective nucleotide, such as different types of base-pairing, stacking, and phosphate-backbone interactions. We find that the RNA SHAPE signal is strongly correlated with cis-Watson-Crick/Watson-Crick base-pairing and is to a remarkable degree not dependent on other structural properties with the exception of stacking. We subsequently generated probabilistic models that estimate the likelihood that a residue with a given SHAPE score participates in base-pairing. We show that several models that take SHAPE scores of adjacent residues into account perform better in predicting base-pairing compared with individual SHAPE scores. This underscores the context sensitivity of SHAPE and provides a framework for an improved interpretation of the response of RNA to chemical modification. 相似文献
142.
Procházková J Kubala L Kotasová H Gudernová I Šrámková Z Pekarová M Sarkadi B Pacherník J 《Free radical research》2011,45(7):779-787
Intracellular production of reactive oxygen species (ROS) plays an important role in the control of cell physiology. For the assessment of intracellular ROS production, a plethora of fluorescent probes is commonly used. Interestingly, chemical structures of these probes imply they could be substrates of plasma membrane efflux pumps, called ABC transporters. This study tested whether the determination of intracellular ROS production and mitochondrial membrane potential by selected fluorescent probes is modulated by the expression and activity of ABC transporters. The sub-clones of the HL-60 cell line over-expressing MDR1, MRP1 and BCRP transporters were employed. ROS production measured by luminol- and L-012-enhaced chemiluminescence and cytochrome c reduction assay showed similar levels of ROS production in all the employed cell lines. It was proved that dihydrorhodamine 123, dihexiloxocarbocyanine iodide, hydroethidine, tetrachloro-tetraethylbenzimidazolocarbo-cyanine iodide and tetramethylrhodamine ethyl ester perchlorate are substrates for MDR1; dichlorodihydrofluoresceine, hydroethidine and tetramethylrhodamine ethyl ester perchlorate are substrates for MRP1; dichlorodihydrofluoresceine, dihydrorhodamine 123, hydroethidine and tetrachloro-tetraethylbenzimidazolocarbo-cyanine iodide are substrates for BCRP. Thus, the determination of intracellular ROS and mitochondrial potential by the selected probes is significantly altered by ABC transporter activities. The activity of these transporters must be considered when employing fluorescent probes for the assessment of ROS production or mitochondrial membrane potential. 相似文献
143.
Long-Term Tree Cover Dynamics in a Pinyon-Juniper Woodland: Climate-Change-Type Drought Resets Successional Clock 总被引:1,自引:0,他引:1
Woody vegetation has expanded in coverage over the past century in many places globally, exemplified by pinyon-juniper changes
in the Southwestern United States. Extreme drought is one of the few non-management drivers besides fire that might reverse
such cover changes, but this has not been well documented. Here, we assess 68 years of tree cover dynamics across an elevation
gradient of a pinyon-juniper woodland using aerial photographs (1936 and 1959) and QuickBird imagery (2004). Canopy cover
increased 32% from 1936 to the onset of a major drought (2002). The largest relative increase in canopy cover occurred from
1936 to 1959 at the higher elevations, but these gains were eliminated by fires occurring from 1959 to 2002, during which
time lower elevations with low canopy cover exhibited the greatest relative increases. The 2002–2004 drought reduced canopy
cover by 55%, which eliminated gains in cover that occurred since 1936. Relative tree cover loss was highest at low elevations
with low tree cover, but absolute tree cover loss was greater in areas of high tree cover, which increased with elevation.
The loss of more than half of the canopy cover during a 2-year drought period was much greater than losses due to fire or
possible increases due to historic land use (for example, grazing). These results suggest that regional-scale climatic influences
may be more important than land use legacies in controlling tree cover of these and perhaps other semiarid woodlands over
longer time scales—notable given that similar episodes of tree mortality are projected in coming decades with climate change. 相似文献
144.
145.
Vlkova M Rohousova I Drahota J Stanneck D Kruedewagen EM Mencke N Otranto D Volf P 《PLoS neglected tropical diseases》2011,5(10):e1344
Background
Phlebotomine sand flies are blood-sucking insects that can transmit Leishmania parasites. Hosts bitten by sand flies develop an immune response against sand fly salivary antigens. Specific anti-saliva IgG indicate the exposure to the vector and may also help to estimate the risk of Leishmania spp. transmission. In this study, we examined the canine antibody response against the saliva of Phlebotomus perniciosus, the main vector of Leishmania infantum in the Mediterranean Basin, and characterized salivary antigens of this sand fly species.Methodology/Principal Findings
Sera of dogs bitten by P. perniciosus under experimental conditions and dogs naturally exposed to sand flies in a L. infantum focus were tested by ELISA for the presence of anti-P. perniciosus antibodies. Antibody levels positively correlated with the number of blood-fed P. perniciosus females. In naturally exposed dogs the increase of specific IgG, IgG1 and IgG2 was observed during sand fly season. Importantly, Leishmania-positive dogs revealed significantly lower anti-P. perniciosus IgG2 compared to Leishmania-negative ones. Major P. perniciosus antigens were identified by western blot and mass spectrometry as yellow proteins, apyrases and antigen 5-related proteins.Conclusions
Results suggest that monitoring canine antibody response to sand fly saliva in endemic foci could estimate the risk of L. infantum transmission. It may also help to control canine leishmaniasis by evaluating the effectiveness of anti-vector campaigns. Data from the field study where dogs from the Italian focus of L. infantum were naturally exposed to P. perniciosus bites indicates that the levels of anti-P. perniciosus saliva IgG2 negatively correlate with the risk of Leishmania transmission. Thus, specific IgG2 response is suggested as a risk marker of L. infantum transmission for dogs. 相似文献146.
Horničáková M Kohoutová J Schlagnitweit J Wohlschlager C Ettrich R Fiala R Schoefberger W Müller N 《Biomolecular NMR assignments》2011,5(2):169-175
PsbQ is one of the extrinsic proteins situated on the lumenal surface of photosystem II (PSII) in the higher plants and green
algae. Its three-dimensional structure was determined by X-ray crystallography with exception of the residues 14–33. To obtain
further details about its structure and potentially its dynamics, we approached the problem by NMR. In this paper we report
1H, 15N, and 13C NMR assignments for the PsbQ protein. The very challenging oligo-proline stretches could be assigned using 13C-detected NMR experiments that enabled the assignments of twelve out of the thirteen proline residues of PsbQ. The identification
of PsbQ secondary structure elements on the basis of our NMR data was accomplished with the programs TALOS+, web server CS23D
and CS-Rosetta. To obtain additional secondary structure information, three-bond HN-Hα J-coupling constants and deviation of experimental 13Cα and 13Cβ chemical shifts from random coil values were determined. The resulting “consensus” secondary structure of PsbQ compares very
well with the resolved regions of the published X-ray crystallographic structure and gives a first estimate of the structure
of the “missing link” (i.e. residues 14–33), which will serve as the basis for the further investigation of the structure,
dynamics and interactions. 相似文献
147.
Gack MU 《Cell host & microbe》2011,10(3):175-177
Tripartite motif (TRIM) proteins are novel players in antiviral innate immunity. In this issue of Cell Host & Microbe,Taylor et?al. (2011) discover the mechanism of action of TRIM79α and show that this interferon-induced protein restricts tick-borne flaviviruses by targeting the viral polymerase for degradation. 相似文献
148.
Kus V Flachs P Kuda O Bardova K Janovska P Svobodova M Jilkova ZM Rossmeisl M Wang-Sattler R Yu Z Illig T Kopecky J 《PloS one》2011,6(11):e27126
Combining pharmacological treatments and life style interventions is necessary for effective therapy of major diseases associated with obesity, which are clustered in the metabolic syndrome. Acting via multiple mechanisms, combination treatments may reduce dose requirements and, therefore, lower the risk of adverse side effects, which are usually associated with long-term pharmacological interventions. Our previous study in mice fed high-fat diet indicated additivity in preservation of insulin sensitivity and in amelioration of major metabolic syndrome phenotypes by the combination treatment using n-3 long-chain polyunsaturated fatty acids (n-3 LC-PUFA) and rosiglitazone, i.e. an anti-diabetic drug of the thiazolidinedione (TZD) family. We investigated here whether pioglitazone, a TZD-drug in clinical use, could elicit the additive beneficial effects when combined with n-3 LC-PUFA. Adult male mice (C57BL/6N) were fed an obesogenic corn oil-based high-fat diet (cHF) for 8 weeks, or randomly assigned to various dietary treatments (i) cHF+F, cHF with n-3 LC-PUFA concentrate replacing 15% of dietary lipids; (ii) cHF+ROSI, cHF with 10 mg rosiglitazone/kg diet; (iii) cHF+F+ROSI; (iv) cHF+PIO, cHF with 50 mg pioglitazone/kg diet; and (v) cHF+F+PIO, or chow-fed. Plasma concentrations of 163 metabolites were evaluated using a targeted metabolomics approach. Both TZDs preserved glucose homeostasis and normal plasma lipid levels while inducing adiponectin, with pioglitazone showing better effectiveness. The beneficial effects of TZDs were further augmented by the combination treatments. cHF+F+ROSI but not cHF+F+PIO counteracted development of obesity, in correlation with inducibility of fatty acid β-oxidation, as revealed by the metabolomic analysis. By contrast, only cHF+F+PIO eliminated hepatic steatosis and this treatment also reversed insulin resistance in dietary obese mice. Our results reveal differential effects of rosiglitazone and pioglitazone, unmasked in the combination treatment with n-3 LC-PUFA, and support the notion that n-3 LC-PUFA could be used as add-on treatment to TZDs in order to improve diabetic patient's therapy. 相似文献
149.
Ebner S Lang R Mueller EE Eder W Oeller M Moser A Koller J Paulweber B Mayr JA Sperl W Kofler B 《PloS one》2011,6(12):e27192
Background
Because mitochondria play an essential role in energy metabolism, generation of reactive oxygen species (ROS), and apoptosis, sequence variation in the mitochondrial genome has been postulated to be a contributing factor to the etiology of multifactorial age-related diseases, including cancer. The aim of the present study was to compare the frequencies of mitochondrial DNA (mtDNA) haplogroups as well as control region (CR) polymorphisms of patients with malignant melanoma (n = 351) versus those of healthy controls (n = 1598) in Middle Europe.Methodology and Principal Findings
Using primer extension analysis and DNA sequencing, we identified all nine major European mitochondrial haplogroups and known CR polymorphisms. The frequencies of the major mitochondrial haplogroups did not differ significantly between patients and control subjects, whereas the frequencies of the one another linked CR polymorphisms A16183C, T16189C, C16192T, C16270T and T195C were significantly higher in patients with melanoma compared to the controls. Regarding clinical characteristics of the patient cohort, none of the nine major European haplogroups was associated with either Breslow thickness or distant metastasis. The CR polymorphisms A302CC-insertion and T310C-insertion were significantly associated with mean Breslow thickness, whereas the CR polymorphism T16519C was associated with metastasis.Conclusions and Significance
Our results suggest that mtDNA variations could be involved in melanoma etiology and pathogenesis, although the functional consequence of CR polymorphisms remains to be elucidated. 相似文献150.
Mapping the B-A conformational transition along plasmid DNA 总被引:1,自引:0,他引:1
A simple method is presented to monitor conformational isomerizations along genomic DNA. We illustrate properties of the method with the B-A conformational transition induced by ethanol in linearized pUC19 plasmid DNA. At various ethanol concentrations, the DNA was irradiated with ultraviolet light, transferred to a restriction endonuclease buffer and the irradiated DNA was cleaved by 17 restriction endonucleases. The irradiation damaged DNA and the damage blocked the restrictase cleavage. The amount of uncleaved, i.e. damaged, DNA depended on the concentration of ethanol in a characteristic S-shape way typical of the cooperative B-A transition. The transition beginning and midpoint were determined for each restriction endonuclease. These data map the B-A transition along the whole polylinker of pUC19 DNA and six evenly distributed recognition sequences within the rest of the plasmid. The transition midpoints fell within the B-A transition region of the plasmid simultaneously determined by CD spectroscopy. The present method complements the previous methods used to study the B-A transition. It can be employed to analyze multikilobase regions of genomic DNA whose restriction endonuclease cleavage fragments can be separated and quantified on agarose gels. 相似文献