Journal of Applied Phycology - Pharmaceuticals (mostly antibiotics and non-steroid anti-inflammatory drugs) are hazardous micropollutants (HMP). Incomplete degradation of the HMP leads to their... 相似文献
EGFP is a fluorescent tag extensively used in biological and biomedical research. Over the years many researches have gathered collections of cell lines bearing specific EGFP-tagged proteins. Despite its popularity some photochemical properties of EGFP remain undocumented and unused. We report on so far unexplored lifetime photoconversion of EGFP usable in FLIM.
Methods
Fluorescence lifetime imaging and spectral FLIM has been used for characterization of the EGFP photoconversion and protein tracking.
Result
Our data suggest that EGFP can be permanently photoconverted to a short-fluorescence-lifetime form (PC-EGFP) by intense blue irradiation. PC-EGFP cannot be reverted back by 405?nm light and exhibits the same spectral emission properties with blue-shifted absorption compared to the unconverted EGFP. Fluorescence of PC-EGFP is pH-independent and the photoconversion efficiency decreases with the solvent viscosity. Utilization of the EGFP photoconversion was demonstrated by tracking of a nucleophosmin mutant in live HEK-293?T cells during its cytoplasm-nuclear relocalization induced by Leptomycin B.
Conclusions
Besides potential FLIM artifacts caused by an unintended EGFP photoconversion, the controlled photoconversion turns EGFP to an excellent tool for kinetic FLIM applications. Since the photoconversion occurs in the lifetime domain, PC-EGFP can be easily distinguished from the unconverted tag by time-resolved detection while all other spectral channels stay free for multicolor labeling.
General significance
The reported lifetime photoconversion lines up EGFP with other photoconvertible fluorescent proteins with special advantage for fluorescence lifetime imaging where lifetime-photoconvertible labels are scarce. 相似文献
Understanding the responses of vegetation characteristics and soil properties to grazing in different precipitation regimes is useful for the management of rangelands, especially in the arid regions. In northeastern Iran, we studied the responses of vegetation to livestock grazing in three regions with different climates: arid, semiarid, and subhumid. In each region, we selected 6–7 pairwise sampling areas of high versus low grazing intensity and six traits of the present species were recorded on 1 m2 plots—five grazed and five ungrazed in each area. The overall fertility was compared using the dissimilarity analysis, and linear mixed‐effect models were used to compare the individual fertility parameters, functional diversity indices, and species traits between the plots with high and low grazing intensity and between the climatic regions. Both climate and grazing, as well as their interaction, affected fertility parameters, functional diversity indices, and the representation of species traits. Grazing reduced functional evenness, height of the community, the representation of annuals, but increased the community leaf area. In the subhumid region, grazing also reduced functional richness. Further, grazing decreased the share of annual species in the semiarid region and seed mass in the arid region. Larger leaf area and seed mass, smaller height and lower share of annuals were associated with intensive grazing. Species with large LA and seed mass, lower height and perennials can be therefore presumed to tolerate trampling and benefit from high nutrient levels, associated with intensive grazing. By providing a detailed view on the impacts of overgrazing, this study highlights the importance of protection from grazing as an effective management tool for maintaining the pastoral ecosystems. In general, the composition of plant traits across the pastures of northeastern Iran was more affected by intensive grazing than by the differences in climate. 相似文献
PurposeTo evaluate the opportunities of single photon emission tomography/computerized tomography (SPECT-CT) for localization of axillary sentinel lymph nodes (ASLNs) and subsequent radiotherapy planning in women with early breast cancer.Material and methodsIndividual topography of ASLN was determined in 151 women with clinical T1-2N0M0 breast cancer. SPECT-CT visualization of ASLNs was initiated 120 min after intra-peritumoral injection of 99mTc-radiocolloids. Doses absorbed by virtual ASLNs after the whole breast irradiation with standard and extended tangential fields were calculated on a treatment planning station.ResultsSPECT-CT demonstrated a large variability of ASLN localization. They were detected in the central subgroup in 94 (61%) patients, in pectoral – in 77 (51%), and in interpectoral – in 4 (3%) patients. Sentinel lymph nodes “lying on the chest” were revealed in 35 (23%) cases.We found that with standard tangential fields coverage of ASLNs was obtained only in 20% of evaluated women. Extended tangential fields can effectively irradiate ASLNs localized in all axillary sub-regions with the exception of ASLNs “lying on the chest”.ConclusionSPECT-CT mapping of ASLNs in women with cT1-2N0M0 breast cancer reveals their variable localization. This information can be important for planning of radiation treatment in women that underwent breast conserving surgery without an axillary surgery. 相似文献
In the sustained presence of agonist, the opening of P2X7R channel is followed by pore dilatation, which causes an increase in its permeability to larger organic cations, accompanied by receptor sensitization. To explore the molecular mechanisms by which the conductivity and sensitivity are increased, we analyzed the electrophysiological properties and YO‐PRO‐1 uptake of selected alanine mutants in the first and second transmembrane domains of the rat P2X7R. Substitution of residues Y40, F43, G338, and D352 with alanine reduced membrane trafficking, and the D352A was practically non‐functional. The Y40A and F43A mutants that were expressed in the membrane lacked pore dilation ability. Moreover, the Y40A and Y40F displayed desensitization, whereas the Y40W partially recovered receptor function. The G338A/S mutations favored the open state of the channel and displayed instantaneous permeability to larger organic cations. The G338P was non‐functional. The L341A and G345A displayed normal trafficking, current amplitude, and sensitization, but both mutations resulted in a decreased pore formation and dye uptake. These results showed that the increase in P2X7R conductivity and sensitivity is critically dependent on residues Y40 and F43 in the TM1 domain and that the region located at the intersection of TM2 helices controls the rate of large pore opening.
Determining which drivers lead to a specific species assemblage is a central issue in community ecology. Although many processes are involved, plant–plant interactions are among the most important. The phylogenetic limiting similarity hypothesis states that closely related species tend to compete stronger than distantly related species, although evidence is inconclusive. We used ecological and phylogenetic data on alpine plant communities along an environmental severity gradient to assess the importance of phylogenetic relatedness in affecting the interaction between cushion plants and the whole community, and how these interactions may affect community assemblage and diversity. We first measured species richness and individual biomass of species growing within and outside the nurse cushion species, Arenaria tetraquetra. We then assembled the phylogenetic tree of species present in both communities and calculated the phylogenetic distance between the cushion species and its beneficiary species, as well as the phylogenetic community structure. We also estimated changes in species richness at the local level due to the presence of cushions. The effects of cushions on closely related species changed from negative to positive as environmental conditions became more severe, while the interaction with distantly related species did not change along the environmental gradient. Overall, we found an environmental context‐dependence in patterns of phylogenetic similarity, as the interaction outcome between nurses and their close and distantly‐related species showed an opposite pattern with environmental severity. 相似文献