Fluorescence of nitrobenzoxadiazole (NBD)-labeled lipids in model membranes is connected not to lipid mobility but to probe location

Nitrobenzoxadiazole (NBD)-labeled lipids are popular fluorescent membrane probes. However, the understanding of important aspects of the photophysics of NBD remains incomplete, including the observed shift in the emission spectrum of NBD-lipids to longer wavelengths following excitation at the red edge of the absorption spectrum (red-edge excitation shift or REES). REES of NBD-lipids in membrane environments has been previously interpreted as reflecting restricted mobility of solvent surrounding the fluorophore. However, this requires a large change in the dipole moment (Dm) of NBD upon excitation. Previous calculations of the value of Dm of NBD in the literature have been carried out using outdated semi-empirical methods, leading to conflicting values. Using up-to-date density functional theory methods, we recalculated the value of Dm and verified that it is rather small (B2 D). Fluorescence measurements confirmed that the value of REES is B16 nm for 1,2-dioleoyl-sn-glycero-3- phospho-L-serine-N-(NBD) (NBD-PS) in dioleoylphosphatidylcholine vesicles. However, the observed shift is independent of both the temperature and the presence of cholesterol and is therefore insensitive to the mobility and hydration of the membrane. Moreover, red-edge excitation leads to an increased contribution of the decay component with a shorter lifetime, whereas time-resolved emission spectra of NBD-PS displayed an atypical blue shift following excitation. This excludes restrictions to solvent relaxation as the cause of the measured REES and TRES of NBD, pointing instead to the heterogeneous transverse location of probes as the origin of these effects. The latter hypothesis was confirmed by molecular dynamics simulations, from which the calculated heterogeneity of the hydration and location of NBD correlated with the measured fluorescence lifetimes/REES. Globally, our combination of theoretical and experiment-based techniques has led to a considerably improved understanding of the photophysics of NBD and a reinterpretation of its REES in particular.

Crossing frontiers between tourism and demography. An empirical analysis based on European travellers’ behaviour

This paper aims to establish possible tourism demand scenarios of European travellers to Portugal based on the relationship with changing population structures. A combination of the EuROBAROMETER report 370 (“Attitudes of Europeans towards Tourism in 2013”) and the cohort-component method for population projections will allow the development of different possible tourism demand scenarios. Following the European report, individuals who travelled in 2013 were most likely to live in a household with two or more individuals. Thus, if elderly couples are together till later in their life and in better physiological shape, it is possible that the number of elderly individuals travelling for tourism purposes will increase in the near future. If we can expect tourists from developing countries to be younger due to their demographic dynamics than those from developed countries, where the ageing population is growing fast, we can expect that the percentage of the elderly among tourists will increase. Furthermore, the 2013 European report found that the combination of socio-demographic variables, such as, age, population, gender, household dimension, country of residence and trip purpose explained tourism demand scenarios for Portugal, confirming that seniors and families evidence a paramount sense of importance for the destination. In the literature there is a lack of discussion about the effects of demography in the future and the role of an ageing population in tourism demand choice patterns. We aim to contribute to filling this gap. Consequently, we strongly believe that this paper contributes to the literature by introducing a new field of discussion about the importance of demographic changes in shaping travel trends.

Quantification of free auxins in semi-hardwood plant cuttings and microshoots by dispersive liquid–liquid microextraction/microwave derivatization and GC/MS analysis

Several studies have suggested that differences in the natural rooting ability of plant cuttings could be attributed to differences in endogenous auxin levels. Hence, during rooting experiments, it is important to be able to routinely monitor the evolution of endogenous levels of plant hormones. This work reports the development of a new method for the quantification of free auxins in auxin-treated Olea europaea (L.) explants, using dispersive liquid–liquid microextraction (DLLME) and microwave assisted derivatization (MAD) followed by gas chromatography/mass spectrometry (GC/MS) analysis. Linear ranges of 0.5–500 ng mL 1 and 1–500 mg mL 1 were used for the quantification of indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA), respectively. Determined by serial dilutions, the limits of detection (LOD) and quantification (LOQ) were 0.05 ng mL 1 and 0.25 ng mL 1, respectively for both compounds. When using the calibration curve for determination, the LOQ corresponded to 0.5 ng mL 1 (IAA) and 0.5 mg mL 1 (IBA). The proposed method proved to be substantially faster than other alternatives, and allowed free auxin quantification in real samples of semi-hardwood cuttings and microshoots of two olive cultivars. The concentrations found in the analyzed samples are in the range of 0.131–0.342 mg g 1 (IAA) and 20–264 mg g 1 (IBA).

Crossing frontiers between tourism and demography. An empirical analysis based on European travellers’ behaviour

This paper aims to establish possible tourism demand scenarios of European travellers to Portugal based on the relationship with changing population structures. A combination of the EUROBAROMETER report 370 (“Attitudes of Europeans towards Tourism in 2013”) and the cohort-component method for population projections will allow the development of different possible tourism demand scenarios. Following the European report, individuals who travelled in 2013 were most likely to live in a household with two or more individuals. Thus, if elderly couples are together till later in their life and in better physiological shape, it is possible that the number of elderly individuals travelling for tourism purposes will increase in the near future. If we can expect tourists from developing countries to be younger due to their demographic dynamics than those from developed countries, where the ageing population is growing fast, we can expect that the percentage of the elderly among tourists will increase. Furthermore, the 2013 European report found that the combination of socio-demographic variables, such as, age, population, gender, household dimension, country of residence and trip purpose explained tourism demand scenarios for Portugal, confirming that seniors and families evidence a paramount sense of importance for the destination. In the literature there is a lack of discussion about the effects of demography in the future and the role of an ageing population in tourism demand choice patterns. We aim to contribute to filling this gap. Consequently, we strongly believe that this paper contributes to the literature by introducing a new field of discussion about the importance of demographic changes in shaping travel trends.