Quantitative stakeholder-driven assessment of radiation protection issues via a PIANOFORTE online survey.

To enhance stakeholder engagement and foster the inclusion of interests of citizens in radiation protection research, a comprehensive online survey was developed within the framework of the European Partnership PIANOFORTE. This survey was performed in 2022 and presented an opportunity for a wide range of stakeholders to voice their opinions on research priorities in radiation protection for the foreseeable future. Simultaneously, it delved into pertinent issues surrounding general radiation protection. The PIANOFORTE e-survey was conducted in the English language, accommodating a diverse range of participants. Overall, 440 respondents provided their insights and feedback, representing a broad geographical reach encompassing 29 European countries, as well as Canada, China, Colombia, India, and the United States. To assess the outcomes, the Positive Matrix Factorization numerical model was applied, in addition to qualitative and quantitative assessment of individual responses, enabling the discernment of four distinct stakeholder groups with varying attitudes. While the questionnaire may not fully represent all stakeholders due to the limited respondent pool, it is noteworthy that approximately 70% of the participants were newcomers to comparable surveys, demonstrating a proactive attitude, a strong willingness to collaborate and the necessity to continuously engage with stakeholder groups. Among the individual respondents, distinct opinions emerged particularly regarding health effects of radiation exposure, medical use of radiation, radiation protection of workers and the public, as well as emergency and recovery preparedness and response. In cluster analysis, none of the identified groups had clear preferences concerning the prioritization of future radiation protection research topics.

Assessing radiation risk perception by means of a European stakeholder survey.

It is increasingly recognised that stakeholder views can be essential for ascertaining the credibility of those entrusted with protection of the public and workers against radiation risks, the robustness of the approaches to protection and the relevance of research underpinning radiation protection (RP). The CONCERT European Joint Programme of RP research included consideration of stakeholder views. These were evaluated by means of a publicly available survey, translated into 15 languages, to encourage responses from a wide range of European countries. The survey ran in 2017 and received some 1961 responses from many countries, although response rates varied widely between countries. The survey respondents were largely highly educated, with many having a professional connection to RP or the use of radiation in medicine or industry. Survey results indicated a high level of scientific/technical knowledge relevant to RP and indicated a general trust of most actors involved in the RP field, perhaps unsurprisingly given the nature of the sampled population. Most expressed a reasonable level of satisfaction with the information available to them on radiation risk, but there is clearly room for improvement. Additionally, the survey identified potential training needs amongst the groups who responded. It is concluded that, while the survey results are limited by the non-representativeness of the respondents by comparison with the population of the European Union as a whole, it has been successful in gaining insights into areas where communication could be improved, where professional training gaps are present and where research could help to build wider trust in RP.

Uncertainties in the use of concentration ratios for modelling NORM waste sites.

The activity concentrations of 238U, 226Ra and 210Pb were modelled in Pinus sylvestris (Scots pine trees) on a uniform CaF2 sludge heap in Belgium. The aim of this work is to enhance the knowledge of how transfer factors behave in NORM landfills. The simplest possible model in radioecology is used, which is based on Concentration Ratios (CR-s) measured in equilibrium and activity concentrations of the above-mentioned radionuclides measured in the substrate where pine trees grow. Two alternative CR-s were used: (1) international CR compilations by the IAEA (2014) and (2) CR-s specifically determined for pine trees studied in British Columbia (Mahon and Mathews, 1983). Both CR-s were applied assuming lognormal distributions fitted from data reported in the literature. The results were compared with activity concentrations measured in trees sampled on-site. Modelled concentrations match the measured ones best in the case of 238U. For the studied NORM waste site, the approach using generic IAEA concentration ratios does not fulfill the conservatism requirement in the cases of 238U and 226Ra, as the concentration of radionuclides in trees is underestimated. On the other hand, the ratios from Mahon and Mathews, (1983) produce wide distributions, ensuring conservatism due to larger CR-s. The measured concentrations are narrowly distributed in general, which can be expected on a small sampling site on a uniform substrate. The generic approach outlined here is practical but, as a result of the uniqueness of the site considered, should be applied cautiously in other NORM situations.

Long-term modelling of fly ash and radionuclide emissions as well as deposition fluxes due to the operation of large oil shale-fired power plants.

Two of the world's largest oil shale-fired power plants (PPs) in Estonia have been operational over 40 years, emitting various pollutants, such as fly ash, SOx, NOx, heavy metals, volatile organic compounds as well as radionuclides to the environment. The emissions from these PPs have varied significantly during this period, with the maximum during the 1970s and 1980s. The oil shale burned in the PPs contains naturally occurring radionuclides from the 238U and 232Th decay series as well as 40K. These radionuclides become enriched in fly ash fractions (up to 10 times), especially in the fine fly ash escaping the purification system. Using a validated Gaussian-plume model, atmospheric dispersion modelling was carried out to determine the quantity and a real magnitude of fly ash and radionuclide deposition fluxes during different decades. The maximum deposition fluxes of volatile radionuclides (210Pb and 210Po) were around 70 mBq m-2 d-1 nearby the PPs during 1970s and 1980s. Due to the reduction of burned oil shale and significant renovations done on the PPs, the deposition fluxes were reduced to 10 mBq m-2 d-1 in the 2000s and down to 1.5 mBq m-2 d-1 in 2015. The maximum deposition occurs within couple of kilometers of the PPs, but the impacted area extends to over 50 km from the sources. For many radionuclides, including 210Po, the PPs have been larger contributors of radionuclides to the environment via atmospheric pathway than natural sources. This is the first time that the emissions and deposition fluxes of radionuclides from the PPs have been quantified, providing the information about their radionuclide deposition load on the surrounding environment during various time periods.

Pb-210 and Po-210 atmospheric releases via fly ash from oil shale-fired power plants.

During high temperature processes in the furnace volatile and semi-volatile elements and radionuclides are partially emitted to the environment, depending on their chemical form in the original fuel, the technological set-up of the combustion system, and the prevailing combustion conditions. Two of the world's largest oil shale-fired power plants (PPs) have been operational in Estonia from the 1960s, during which time creation of significant environmental emissions and waste containing naturally occurring radionuclides has occurred. Pb-210 and 210Po are considered natural radionuclides with the highest emission rates from PPs and possess elevated potential radiation exposure risks to humans and the environment. These radionuclides have the highest activity concentration values in fine ash fractions, especially in fractions remaining below 2.5 µm. To determine the activity concentrations of 210Pb and 210Po in the PPs' outlet, sampling was conducted from boilers operating on pulverized fuel (PF) technology with novel integrated desulphurization (NID) system and bag filters as well as with electrostatic precipitators (ESPs). The 210Pb and 210Po activity concentrations remained around 300 Bq kg-1 for the NID system compared to 60-80 Bq kg-1 in the ESP system. The dominant ash fraction in both systems was PM2.5, constituting over 50% of the fly ash mass collected from the outlet. The authors estimate that the total atmospherically emitted activity for the modernized PPs remains dominantly below 1% of the activity that is inserted via fuel. The implementation of higher efficiency purifications systems has significantly reduced the negative effect of these PPs. Based on annually emitted fly ash and boilers' working hours, the 210Pb and 210Po activity released relative to energy production were up to 68.3 kBq GWhel-1 for 210Pb and 64.6 kBq GWhel-1 for 210Po. These values are 1 to 2 orders of magnitude lower compared to the situation in the 1980s. These findings represent the first publicly available quantitative results estimating the 210Po emissions from large oil shale-fired PPs.

Gaussian beam reflection and refraction by a spherical or parabolic surface: comparison of vectorial-law calculation with lens approximation.

A ray-tracing approach is used to demonstrate efficient application of the vectorial laws of reflection and refraction to computational optics problems. Both the full width at half-maximum (fwhm) and offset of Gaussian beams resulting from off-center reflection and refraction are calculated for spherical and paraboloidal surfaces of revolution. It is found that the magnification and displacement depend nonlinearly on the miscentering. For these geometries, the limits of accuracy of the lens approximation are examined quantitatively. In contrast to the ray-tracing solution, this paraxial approximation would predict a magnification of a beam's fwhm that is independent of miscentering, and an offset linearly proportional to the miscentering. The focusing property of paraboloidal surfaces of revolution is also derived in setting up the calculation.

Cataract diagnosis by measurement of backscattered light.

We present a portable optical cataract assessment technology which measures with a circular photodetector the fraction of light scattered backwards by the human eye lens when illuminated by a laser diode. As our signal arises directly from the fundamental pathology-increased scattering in the lens-it directly assesses cataract extent and progression. Initial clinical results in undilated human eyes show device reading correlations in agreement with clinical examination and Scheimpflug photography.

Multiphoton flow cytometry strategies and applications.

A handful of research teams around the world have recently begun to utilize multiphoton techniques in cytometry, especially for in vivo applications. These approaches offer similar enhancements to flow cytometry as the multiphoton phenomenon brought to the field of microscopy at the turn of the 20th century, with at least six advantages over single-photon excitation. Here, we review the published literature on multiphoton cytometry in vivo or in vitro from the initial experiments in 1999 to present. Multiphoton cytometry instrumentation set-ups vary from adapted multiphoton microscopy to a dedicated system, with laser pulse power and repetition rate serving as important variables. Two-beam geometry enables quantitation of cell size. Labeling strategies include conjugated fluorophore targeting, with folate and/or dendrimer platforms. With two-color measurement, ratiometric labeling is also possible, where one dye serves as a trigger to indicate the amount of excitation a cell receives, and another informs of cellular function. With two-color labeling, geometric fluorophore distribution proves important in theory and experiment for detection sensitivity curves and detected event intensity correlation. The main biological achievements to date using this young technology are reviewed, with emphasis on real-time monitoring of minute-by-minute and long-term cell dynamics as well as the clinically significant surveillance of circulating tumor cells. For this goal, minimally invasive two-photon flow cytometry with a fiber probe may overcome the primary issue of sample volume. The technique of multicolor, multiphoton flow cytometry greatly enhances the capabilities of flow cytometry to investigate the dynamics of circulating cells in cancer and other important diseases, and may in the future benefit from advances in microscopy such as super-resolution imaging, coherent control, and bioluminescence.

Microfluidic Droplet Consistency Monitoring and Cell Detection via Laser Excitation.

Microfluidic droplets formed in emulsions are used in a variety of analytical techniques and hold great potential for future scientific and commercial applications. Our experiments merge quantitative quality engineering methods into the microdroplet field. We present a unique microdroplet generation and consistency monitoring system with laser optics excitation and detection. Our setup analyzes each droplet with sub-millisecond signal resolution and single photon accuracy, and is compatible with process control methods. To demonstrate the consistency of microdroplet generation over time, we measure and examine the mean frequency of aqueous plug-shaped droplet (microplug) formation in oil phase, as well as the mean length of plugs, and the interval between consecutive droplets. We also demonstrate the detection of cancer cells encapsulated within aqueous microdroplets in continuous oil phase flow. Two-channel optical monitoring allows for the simultaneous and independent inspection of both microdroplet generation and identification of green fluorescent protein-labelled cancer cells within the droplets. Increased accuracy and consistency are central to many established and developing microfluidic technologies. A systematic, quantitative approach as demonstrated with our experiments may be essential in the development of advanced microfluidic concepts that require exacting reproducibility and would greatly benefit from incorporated automated measurement techniques for process control.

Quantitative differentiation of dyes with overlapping one-photon spectra by femtosecond pulse shaping.

We demonstrate that DiI and Rhodamine B, which are not easily distinguishable to one-photon measurements, can be differentiated and in fact quantified in mixture via tailored two-photon excitation pulses found by a genetic algorithm (GA). A nearly three-fold difference in the ratio of two-photon fluorescence of the two dyes is achieved, without a drop in signal of the favored fluorophore. Implementing an acousto-optic interferometer, we were able to prove that the mechanism of discrimination is second-harmonic tuning by the phase-shaped pulses to the relative maxima and minima of these cross-sections.

Control of two-photon fluorescence of common dyes and conjugated dyes.

We present a comprehensive study of the selective excitation of two-photon fluorescence from various pairs of dyes and dyes in different conjugation states with tailored pulse shapes found with a genetic algorithm (GA). We investigate a number of biologically important dyes, and include dyes conjugated to trastuzumab (Herceptin(R)) and to a poly(amidoamine) dendrimer. We consider in detail the ability of tailored pulse shaping to discriminate dyes with significant spectral overlap. Our procedure for adaptive pulse shaping includes power-law and chirp-scaling checks to prevent trivial convergences. The GA uses a multiplicative fitness parameter in a graded search method that converges on pulse shapes that not only differentiate two-photon processes, but do so in a high signal regime. We consider the results in terms of not only the absolute maximum ratio of discrimination achieved, but also present the evolutionary course of the GA and compare the improvement to a quantitative measure of the noise level. We also implement a time-domain acousto-optic measurement of two-photon excitation cross-section spectra. The results show that the ability to discriminate dyes is determined almost entirely by their differences in two-photon excitation cross section.

Extended cavity laser enhanced two-photon flow cytometry.

We demonstrate enhanced sensitivity in two-photon flow cytometry with an extended cavity laser excitation source. At low power, the home-built 20-MHz oscillator was able to detect a significantly larger fraction, in either phosphate buffered saline (PBS) or whole blood, of green fluorescent protein (GFP)-expressing MCA-207 cells cross-labeled with the membrane-binding lipophilic dye DiD. A geometrical model is used to explain unique features of the signals resulting from the different spatial distribution of DiD and GFP. These unique features include sub-square law scaling of unsaturated two-photon signal, a sigmoidal sensitivity curve for detection under varying powers for cell detection thresholds as low as a single photon, and uncorrelated signal strengths in two detection channels.

Control of the blue fluorescent protein with advanced evolutionary pulse shaping.

We demonstrate optical coherent control of the two-photon fluorescence of the blue fluorescent protein (BFP), which is of interest in investigations of protein-protein interactions. In addition to biological relevance, BFP represents an interesting target for coherent control from a chemical perspective due to its many components of highly nonexponential fluorescence decay and low quantum yield resulting from excited state isomerization. Using a genetic algorithm with a multiplicative (rather than ratiometric) fitness parameter, we are able to control the ratio of BFP fluorescence to second-harmonic generation without a considerable drop in the maximized signal. The importance of linear chirp and power-scaling on the discrimination process is investigated in detail.

Physicochemical properties of blue fluorescent protein determined via molecular dynamics simulation.

Blue fluorescent protein (BFP) is a mutant of green fluorescent protein (GFP), where the chromophore has been modified to shift the emitted fluorescence into the blue spectral region. In this study, MD calculations were performed with the GROMACS simulation package and AMBER force field to investigate the dependence of BFPs physicochemical properties on temperature and applied pressure. The MD approach enabled us to calculate the compressibility of protein itself, separately from the nontrivial contribution of the hydration shell, which is difficult to achieve experimentally. The computed compressibility of BFP (3.94 x10(-5) MPa(-1)) is in agreement with experimental values of globular proteins. The center-of-mass diffusion coefficient of BFP and its dependence on temperature and pressure, which plays an important role in its application as a probe for intracellular liquid viscosity measurement, was calculated and found to be in good agreement with photobleaching recovery experimental data. We have shown that decreased temperature as well as applied pressure increases the water viscosity, but the concomitant decrease of the BFP diffusion coefficient behaves differently from Stokes-Einstein formula. It is shown that the number of hydrogen bonds around the protein grows with pressure, which explains the aforementioned deviation. Pressure also reduces root mean square (RMS) fluctuations, especially those of the most flexible residues situated in the loops. The analysis of the RMS fluctuations of the backbone Calpha atoms also reveals that the most rigid part of BFP is the center of the beta-barrel, in accord with temperature B factors obtained from the Protein Data Bank.

Electrowetting of ionic liquids.

We have successfully demonstrated that imidazolium- and pyrrolidinium-based commercial room-temperature ionic liquids can electrowet (with a dc voltage) a smooth fluoropolymer (Teflon AF1600) surface. Qualitatively, the process is analogous to the electrowetting of aqueous electrolyte solutions: the contact angle versus voltage curve has a parabolic shape which saturates at larger voltages (positive or negative). On the other hand we observed several peculiarities: (i) the efficiency is significantly lower (by about an order of magnitude); (ii) the influence of the bulky cation is larger and the importance of the smaller anion is lesser, especially with respect to electrowetting saturation; (iii) there is an asymmetry in the saturation contact angles found for positive and negative voltages. The asymmetry may be correlated with the cation-anion asymmetry of the ionic liquids. The low efficiency may be caused by the presence of water and other impurities in these commercial materials.

Vibrations-determined properties of green fluorescent protein.

The physicochemical characteristics of the green fluorescent protein (GFP), including the thermodynamic properties (entropy, enthalpy, Gibbs' free energy, heat capacity), normal mode vibrations, and atomic fluctuations, were investigated. The Gaussian 03 computational chemistry program was employed for normal mode analysis using the AMBER force field. The thermodynamic parameters and atomic fluctuations were then calculated from the vibrational eigenvalues (frequencies) and eigenvectors. The regions of highest rigidity were shown to be the beta-sheet barrel with the central alpha-helix, which bears the chromophore. The most flexible parts of the GFP molecule were the outlying loops that cover the top and bottom of the beta-barrel. This way, the balance between rigidity and flexibility is maintained, which is the optimal relationship for protein stability in terms of Gibbs' free energy. This dual-schemed structure satisfies the requirements for GFP function. In this sense, the structure of GFP resembles a nanoscale drum: a stiff cylinder with flexible vibrating end(s).

Reversible switching of high-speed air-liquid two-phase flows using electrowetting-assisted flow-pattern change.

This work is the first demonstration of electrical modulation of surface energy to reversibly switch dynamic high-speed gas-liquid two-phase microfluidic flow patterns. Manipulation of dynamic two-phase systems with continuous high-speed flows is complex and interesting due to the multiple types of forces that need to be considered. Here, distinct stable flow patterns are formed through a multipronged approach: both surface tension forces generated by surface chemistry modulation as well as viscous and inertial forces produced by fluid flows are employed. The novel fluidic actuation mechanism provides insights into better understanding microscale two-phase flow dynamics and offers new opportunities for the development of two-phase biochemical microsystems that are mechanically simple and operational at high speeds.

Micro- and nanotechnologies for studying cellular function.

The study of complex biological systems requires methods to perturb the system in complex yet controlled ways to elucidate mechanisms and dynamic interactions, and to recreate in vivo conditions in flexible in vitro set-ups. This paper reviews recent advances in the use of micro- and nanotechnologies in the study of complex biological systems and the advantages they provide in these two areas. Particularly useful for controlling the chemical and mechanical microenvironments of cells is a set of techniques called soft lithography, whereby elastomeric materials are used to transfer and generate micro- and nanoscale patterns. Examples of some of the capabilities of soft lithography include the use of elastomeric stamps to generate micropatterns of protein and the use of elastomeric channels to localize chemicals with subcellular spatial resolutions. These types of biological micro- and nanotechnologies combined with mathematical modeling will propel our understandings of cellular and subcellular physiology to new heights.