Research at the European Synchrotron Radiation Facility medical beamline.

The application of synchrotron radiation in medical research has become a mature field of research at synchrotron facilities worldwide. In the relatively short time that synchrotrons have been available to the scientific community, their characteristic beams of UV and X-ray radiation have been applied to virtually all areas of medical science which use ionizing radiation. The ability to tune intense monochromatic beams over wide energy ranges differentiates these sources from standard clinical and research tools. At the European Synchrotron Radiation Facility (Grenoble, France), a major research facility is operational on an advanced wiggler radiation beamport, ID17. The beamport is designed to carry out a broad range of research ranging from cell radiation biology to in vivo human studies. Medical imaging programs at ID17 include transvenous coronary angiography, computed tomography, mammography and bronchography. In addition, a major research program on microbeam radiation therapy is progressing. This paper will present a very brief overview of the beamline and the imaging and therapy programs.     

A set of fortran subroutines for optimizing radiotherapy plans.

Quadratic Programming techniques have been applied to the optimization of radiation field weighting in Radiotherapy planning. Wedge selection has also been included by means of an exhaustive search. The radiation dose at any point in the patient may be constrained to be less than a stated percentage of the tumour dose. The routines have been successfully interfaced into a small computer interactive planning system, but they could represent an even more powerful tool in batch and time sharing systems. Minimum operator intervention is required in their use.     

ELF Noise Fields: A Review

The debate as to whether low-level electromagnetic fields can affect biological systems and in the long term cause health effects has been going on for a long time. Yet the interaction of weak electromagnetic fields (EMF) with living cells, undoubtedly a most important phenomenon, is still not well understood. The exact mechanisms by which the effects are produced have not been identified. Furthermore, it is not possible to clearly define which aspects of an EMF exposure that constitute the “dose.” One of the groups that contributed to solving this problem is the Bioelectromagnetics group at Catholic University of America (CUA), Washington, D.C. Their work has been devoted to investigating the physical parameters that are needed to obtain an effect of EMF exposure on biological systems, and also how to inhibit the effect. This is a review of their work on bioeffects caused by low-level EMF, their dependence on coherence time, constancy, spatial averaging, and also how the effects can be modified by an applied ELF noise magnetic field. The group has been using early chick embryos, and L929 and Daudi cells as their main experimental systems. The review also covers the work of other groups on low-level effects and the inhibition of the effects with an applied noise field. The group at CUA has shown that biological effects can be found after exposure to low-level ELF and RF electromagnetic fields, and when effects are observed, applying an ELF magnetic noise field inhibits the effects. Also, other research groups have tried to replicate the studies from the CUA group, or to apply EMF noise to suppress EMF-induced effects. Replications of the CUA effects have not always been successful. However, in all cases where the noise field has been applied to prevent an observed effect, it has been successful in eliminating the effect.     

Physicochemical optimisation of plasmid delivery by cationic lipids

Non-viral gene therapy is based on the use of plasmid expression vectors and chemical or physical plasmid DNA delivery systems. This review discusses the roles of cationic lipids as vectors for gene transfection, reviews different strategies employed to improve cationic lipids for in vivo use, and provides original results on the physicochemistry of lipoplexes. Cationic lipid/DNA delivery vehicles have evolved considerably since their initial gene transfection experiments. Much work has been carried out to investigate their structure/activity relationships, methods of formulation and physicochemical properties. Further work has also focused on enhancing and prolonging their stability in a physiological environment as well as increasing their site-specific and tissue-specific interactions. Original data presented in this report confirm that cationic lipids associated to DNA form supramolecular lamellar structures, which protect DNA from serum DNAse degradation. The effect of formulation (and hence the size of the particles) on lipoplex in vivo circulation half-life and biodistribution is also discussed. A list of abbreviations can be found at the end of the review.     

Secondary structure of cell-penetrating peptides controls membrane interaction and insertion

The clinical use of efficient therapeutic agents is often limited by the poor permeability of the biological membranes. In order to enhance their cell delivery, short amphipathic peptides called cell-penetrating peptides (CPPs) have been intensively developed for the last two decades. CPPs are based either on protein transduction domains, model peptide or chimeric constructs and have been used to deliver cargoes into cells through either covalent or non-covalent strategies. Although several parameters are simultaneously involved in their internalization mechanism, recent focuses on CPPs suggested that structural properties and interactions with membrane phospholipids could play a major role in the cellular uptake mechanism. In the present work, we report a comparative analysis of the structural plasticity of 10 well-known CPPs as well as their ability to interact with phospholipid membranes. We propose a new classification of CPPs based on their structural properties, affinity for phospholipids and internalization pathways already reported in the literature.     

Recent Progress in the Development of Anode Materials for Solid Oxide Fuel Cells

The field of research into solid oxide fuel cell (SOFC) anode materials has been rapidly moving forward. In the four years since the last in‐depth review significant advancements have been made in the reduction of the operating temperature and improvement of the performance of SOFCs. This progress report examines the developments in the field and looks to draw conclusions and inspiration from this research. A brief introduction is given to the field, followed by an overview of the principal previous materials. A detailed analysis of the developments of the last 4 years is given using a selection of the available literature, concentrating on metal‐fluorite cermets and perovskite‐based materials. This is followed by a consideration of alternate fuels for use in SOFCs and their associated problems and a short discussion on the effect of synthesis method on anode performance. The concluding remarks compile the significant developments in the field along with a consideration of the promise of future research. The recent progress in the development of anode materials for SOFCs based on oxygen ion conducting electrolytes is reviewed.     

Current knowledge of Chytridiomycota diversity in Northern Europe and future research needs

Chytridiomycota is the most species-rich phylum of basal lineage fungi involved in vital processes in both terrestrial and aquatic ecosystems. Still, the diversity and richness of this group remains cryptic. In Northern Europe, few species have been recorded despite the numerous intact lake systems covering this region. Recent classifications of early diverging fungal lineages differ considerably on the diversity of chytrid species and their taxonomic placement. Here, we present the current knowledge of the Chytridiomycota diversity within Northern Europe by using the Global Biodiversity Information Facility (GBIF) data and compare how this diversity distributes across two recently proposed classification systems. Furthermore, we illustrate how various sampling types are influencing the recorded classification levels. Lastly, we discuss how metabarcoding has contributed to the overall understanding of the Chytridiomycota diversity in revealing the “dark matter fungi”, and we point out future research needs in the field of aquatic mycology in the Nordic region.     

FLUKA simulation of target fragmentation in proton therapy

In proton therapy, secondary fragments are created in nuclear interactions of the beam with the target nuclei. The secondary fragments have low kinetic energies and high atomic numbers as compared to primary protons. Fragments have a high LET and deposit all their energy close to the generation point. For their characteristics, secondary fragments can alter the dose distribution and lead to an increase of RBE for the same delivered physical dose. Moreover, the radiobiological impact of target fragmentation is significant mostly in the region before the Bragg peak, where generally healthy tissues are present, and immediately after Bragg peak. Considering the high biological impact of those particles, especially in the case of healthy tissues or organs at risk, the inclusion of target fragmentation processes in the dose calculation of a treatment planning system can be relevant to improve the treatment accuracy and for this reason it is one of the major tasks of the MoVe IT project.In this study, Monte Carlo simulations were employed to fully characterize the mixed radiation field generated by target fragmentation in proton therapy. The dose averaged LET has been evaluated in case of a Spread Out Bragg Peak (SOBP). Starting from LET distribution, RBE has been evaluated with two different phenomenological models. In order to characterize the mixed radiation field, the production cross section has been evaluated by means of the FLUKA code. The future development of present work is to generate a MC database of fragments fluence to be included in TPS.     

Effects of UV-B Radiation on Anti-predator Behavior in Three Species of Amphibians

Ultraviolet radiation has been suggested as a possible contributing cause of amphibian declines around the world. Both laboratory and field studies have demonstrated that exposure to ultraviolet radiation can lead to increased mortality of developing amphibians. Virtually no studies have examined the sub‐lethal effects of ultraviolet on amphibian behavior. In this study, we examine the anti‐predator behavior of three species of amphibians after short‐term exposure to ultraviolet‐B radiation. Toad (Bufo boreas) juveniles that had been exposed to ultraviolet radiation did not respond to chemical extracts from conspecifics and heterospecifics as much as juveniles that had not been exposed. Both newt larvae (Taricha granulosa) that had been exposed to ultraviolet radiation and those that had not been exposed responded to chemical cues from conspecific predators by increasing the amount of time spent in shelter. Frog tadpoles (Rana cascadae) that had been exposed to ultraviolet radiation did not reduce their movement in response to chemical cues from predators as much as tadpoles that had not been exposed. These results indicate that ultraviolet exposure may have important sub‐lethal effects in amphibians that could adversely effect their fitness.     

Theoretical and Experimental Study of Surface Plasmon Radiation Force on Micrometer-Sized Spheres

We report theoretical and experimental study of surface plasmon (SP) radiation force on micrometer-sized polystyrene latex spheres with different radii. Two theoretical methods are introduced to numerically investigate the SP sphere system: the arbitrary beam theory for electromagnetic field distribution and the generalized Lorenz–Mie scattering theory for radiation force calculation. Transport velocity of the spheres in water is tested experimentally and compared with numerical results. Our work also shows effective coupling from SP field to micrometer-sized spheres, with high consistency between experiment and calculation, which has significant potential in future applications.     

Failla Memorial Lecture. From beans to genes--back to the future.

Developments in radiation biology have inevitably paralleled the evolution of systems and end points in the wider field of biology. I review the development of three areas that have interested me during the 35 years that I have been involved with radiation research. (1) The dose-rate effect. My first graduate student, Joel Bedford, and I demonstrated the dependence of mammalian cell killing on the dose rate at which gamma rays are delivered. These studies led to the recent development of pulsed low-dose-rate brachytherapy and the design of a new machine for clinical use. (2) Hypoxic radiosensitizers and bioreductive drugs. The hypothesis that the presence of foci of hypoxic cells in human tumors could limit their curability by gamma rays led to the development of nitromidazales that preferentially sensitize hypoxic cells. Our contributions included the observation that misonidazole was also cytotoxic to hypoxic cells, that sensitization was increased by prolonged preincubation, and that the mechanism involved depletion of thiols. More recently we have collaborated with Ged Adams and Bob Sutherland in the development of a new generation of hypoxic cell cytotoxins. In particular, we have been concerned with the assessment of oncogenicity in relation to drug structure. (3) Oncogenic transformation in vitro. The use of in vitro assays for oncogenic transformation has represented a major interest for this laboratory largely due to collaborations first with Carmia Borek, and more recently with Richard Miller and Tom Hei. We published the first dose-response curves for gamma rays, and more recently for a range of neutron energies and for charged particles of defined LET. Radiation-induced oncogenic transformation in C3H 10T1/2 cells has been shown to be due to a dominant acting transforming gene, which has been isolated and is in the process of being characterized and sequenced by Greg Freyer.     

Perspectives on the use of an endogenous gene target in studies of mutational specificity

Mutational spectra have become increasingly important tools in generating a molecular level understanding of mutagenesis in mammalian cells. The work in this field has primarily involved the use of shuttle vector systems although some work has also been reported using endogenous cellular genes as mutational targets. In this communication we discuss the relative utility of these two approaches. We specifically focus on UV light-induced mutagenesis since this agent has been studied in both types of system. We conclude that shuttle vector and endogenous gene studies of mutagenesis are highly complementary, each possessing unique advantages.     

Automated selective dissociation of cells from different regions of multicellular spheroids

Summary In this report we describe a new apparatus which has been developed for the automated selective dissociation of multicellular spheroids into fractions of viable cells from different locations in the spheroid. This device is based on the exposure of spheroids to a 0.25% solution of trypsin under carefully controlled conditions, such that the cells are released from the outer spheroid surface in successive layers. Study of the spheroid size, number of cells per spheroid, and sections through the spheroid with increasing exposure to trypsin demonstrate the effectiveness of this technique. The technique has been successfully used on spheroids from five different cell lines over a wide range of spheroid diameters. We also present data detailing the effect of varying the dissociation temperature, the mixing speed, the trypsin concentration, and the number of spheroids being dissociated. The new apparatus has several advantages over previous selective dissociation methods and other techniques for isolating cells from different regions in spheroids, including: a) precise control over dissociation conditions, improving reproducibility; b) short time to recover cell fractions; c) ability to isolate large numbers of cells from many different spheroid locations; d) use of common, inexpensive laboratory equipment; and e) easy adaptability to new cell lines or various spheroid sizes. Applications of this method are demonstrated, including the measurement of nutrient consumption rates, regrowth kinetics, and radiation survivals of cells from different spheroid regions. This work was supported by grants CA-36535, CA-22585, and RR-02845 from the National Institutes of Health, Bethesda, MD, the National Flow Cytometry Resource (NIH grant RR-01315), and by the Department of Energy, Washington, DC.     

Radiation biology of mosquitoes

There is currently renewed interest in assessing the feasibility of the sterile insect technique (SIT) to control African malaria vectors in designated areas. The SIT relies on the sterilization of males before mass release, with sterilization currently being achieved through the use of ionizing radiation. This paper reviews previous work on radiation sterilization of Anopheles mosquitoes. In general, the pupal stage was irradiated due to ease of handling compared to the adult stage. The dose-response curve between the induced sterility and log (dose) was shown to be sigmoid, and there was a marked species difference in radiation sensitivity. Mating competitiveness studies have generally been performed under laboratory conditions. The competitiveness of males irradiated at high doses was relatively poor, but with increasing ratios of sterile males, egg hatch could be lowered effectively. Males irradiated as pupae had a lower competitiveness compared to males irradiated as adults, but the use of partially-sterilizing doses has not been studied extensively. Methods to reduce somatic damage during the irradiation process as well as the use of other agents or techniques to induce sterility are discussed. It is concluded that the optimal radiation dose chosen for insects that are to be released during an SIT programme should ensure a balance between induced sterility of males and their field competitiveness, with competitiveness being determined under (semi-) field conditions. Self-contained ⁶⁰Co research irradiators remain the most practical irradiators but these are likely to be replaced in the future by a new generation of high output X ray irradiators.     

Cancer predictive value of cytogenetic markers used in occupational health surveillance programs: a report from an ongoing study by the European Study Group on Cytogenetic Biomarkers and Health

The cytogenetic endpoints in peripheral blood lymphocytes: chromosomal aberrations (CA), sister chromatid exchange (SCE) and micronuclei (MN) are established biomarkers of exposure for mutagens or carcinogens in the work environment. However, it is not clear whether these biomarkers also may serve as biomarkers for genotoxic effects which will result in an enhanced cancer risk. In order to assess this problem, Nordic and Italian cohorts were established, and preliminary results from these two studies indicated a predictive value of CA frequency for cancer risk, whereas no such associations were observed for SCE or MN. A collaborative study between the Nordic and Italian research groups, will enable a more thorough evaluation of the cancer predictivity of the cytogenetic endpoints. We here report on the establishment of a joint data base comprising 5271 subjects, examined 1965–1988 for at least one cytogenetic biomarker. Totally, 3540 subjects had been examined for CA, 2702 for SCE and 1496 for MN. These cohorts have been followed-up with respect to subsequent cancer mortality or cancer incidence, and the expected values have been calculated from rates derived from the general populations in each country. Stratified cohort analyses will be performed with respect to the levels of the cytogenetic biomarkers. The importance of potential effect modifiers such as gender, age at test, and time since test, will be evaluated using Poisson regression models. The remaining two potential effect modifiers, occupational exposures and smoking, will be assessed in a case-referent study within the study base.     

Light‐Driven BiVO4–C Fuel Cell with Simultaneous Production of H2O2

Photoelectrochemical (PEC) systems have been researched for decades due to their great promise to convert sunlight to fuels. The majority of the research on PEC has been using light to split water to hydrogen and oxygen, and its performance is limited by the need of additional bias. Another research direction on PEC using light, is to decompose organic materials while producing electricity. In this work, the authors report a new type of unassisted PEC system that uses light, water and oxygen to simultaneously produce electricity and hydrogen peroxide (H₂O₂) on both the photoanode and cathode, which is essentially a light‐driven fuel cell with H₂O₂ as the main product at the two electrodes, meanwhile achieving a maximum power density of 0.194 mW cm^‐2, an open circuit voltage of 0.61 V, and a short circuit current density of 1.09 mA cm^‐2. The electricity output can be further used as a sign for cell function when accompanied by a detector such as a light‐emitting diode (LED) light or a multimeter. This is the first work that shows H₂O₂ two‐side generation with a strict key factors study of the system, with a clear demonstration of electricity output ability using low‐cost earth abundant materials on both sides, which represents an exciting new direction for PEC systems.     

Molecular dynamics simulations for the prediction of the dielectric spectra of alcohols,glycols and monoethanolamine

The response of molecular systems to electromagnetic radiation in the microwave region (0.3–300 GHz) has been principally studied experimentally, using broadband dielectric spectroscopy. However, relaxation times corresponding to reorganisation of molecular dipoles due to their interaction with electromagnetic radiation at microwave frequencies are within the scope of modern molecular simulations. In this work, fluctuations of the total dipole moment of a molecular system, obtained through molecular dynamics simulations, are used to determine the dielectric spectra of water, a series of alcohols and glycols, and monoethanolamine. Although the force fields employed in this study have principally been developed to describe thermodynamic properties, most them give fairly good predictions of this dynamical property for these systems. However, the inaccuracy of some models and the long simulation times required for the accurate estimation of the static dielectric constant can sometimes be problematic. We show that the use of the experimental value for the static dielectric constant in the calculations, instead of the one predicted by the different models, yields satisfactory results for the dielectric spectra, and hence the heat absorbed from microwaves, avoiding the need for extraordinarily long simulations or re-calibration of molecular models.     

Penetration and effects of solar UV-B on phytoplankton and macroalgae

The effects of short wavelength solar radiation on aquatic ecosystems were studied in several marine and freshwater systems. The spectral distribution and the penetration of solar radiation into different water types (coastal and oceanic waters of the Baltic Sea, North Sea, Atlantic and Mediterranean) were investigated. Penetration of short wavelength solar radiation strongly depends on the content of dissolved and particulate substances as well as the concentration of phytoplankton. The primary producers often show a typical vertical distribution within the euphotic zone and are reached as well as affected by the penetrating UV-B radiation. The effect of this radiation was both determined in phytoplankton and macroalgae. Measuring pulse amplitude modulated (PAM) fluorescence indicated that major biomass producers were severely inhibited by surface radiation and even impaired at their natural growth site. Likewise, photosynthetic oxygen production was affected by extended exposure to solar radiation.