Towards an ecological modelling approach for assessing ionizing radiation impact on wildlife populations.

The emphasis of the international system of radiological protection of the environment is to protect populations of flora and fauna. Throughout the MODARIA programmes, the United Nations' International Atomic Energy Agency (IAEA) has facilitated knowledge sharing, data gathering and model development on the effect of radiation on wildlife. We present a summary of the achievements of MODARIA I and II on wildlife dose effect modelling, extending to a new sensitivity analysis and model development to incorporate other stressors. We reviewed evidence on historical doses and transgenerational effects on wildlife from radioactively contaminated areas. We also evaluated chemical population modelling approaches, discussing similarities and differences between chemical and radiological impact assessment in wildlife. We developed population modelling methodologies by sourcing life history and radiosensitivity data and evaluating the available models, leading to the formulation of an ecosystem-based mathematical approach. This resulted in an ecologically relevant conceptual population model, which we used to produce advice on the evaluation of risk criteria used in the radiological protection of the environment and a proposed modelling extension for chemicals. This work seeks to inform stakeholder dialogue on factors influencing wildlife population responses to radiation, including discussions on the ecological relevance of current environmental protection criteria. The area of assessment of radiation effects in wildlife is still developing with underlying data and models continuing to be improved. IAEA's ongoing support to facilitate the sharing of new knowledge, models and approaches to Member States is highlighted, and we give suggestions for future developments in this regard.

New

Precise antineutrino measurements are very sensitive to proper background characterization. We present an improved measurement of the ^{13}C(α,n)^{16}O reaction cross section which constitutes significant background for large ν[over ¯] detectors. We greatly improve the precision and accuracy by utilizing a setup that is sensitive to the neutron energies while making measurements of the excited state transitions via secondary γ-ray detection. Our results shows a 54% reduction in the background contributions from the ^{16}O(3^{-},6.13 MeV) state used in the KamLAND analysis.

New Measurement of

Carbon and oxygen burning reactions, in particular, ^{12}C+^{12}C fusion, are important for the understanding and interpretation of the late phases of stellar evolution as well as the ignition and nucleosynthesis in cataclysmic binary systems such as type Ia supernovae and x-ray superbursts. A new measurement of this reaction has been performed at the University of Notre Dame using particle-γ coincidence techniques with SAND (a silicon detector array) at the high-intensity 5U Pelletron accelerator. New results for ^{12}C+^{12}C fusion at low energies relevant to nuclear astrophysics are reported. They show strong disagreement with a recent measurement using the indirect Trojan Horse method. The impact on the carbon burning process under astrophysical scenarios will be discussed.

The influence of smoking on radiation-induced bystander signal production in esophageal cancer patients.

The relevance of radiation-induced bystander effects in humans is unclear. Much of the existing data relate to cell lines but the effect of bystander signals in complex human tissues is unclear. A phase II clinical study was untaken, where blood sera from 60 patients along with 15 cancer-free volunteers were used to detect whether measurable bystander factor(s) could be found in the blood following high dose rate (HDR) brachytherapy. Overall, there was no significant change in bystander signal production (measured in a human keratinocyte reporter system) before and after one treatment fraction of HDR brachytherapy (p>0.05). Further assessment of patient characteristics and environmental modifiable factors including smoking were also analyzed. Similar to previously published data, samples taken from smokers produced weaker signals compared to non-smokers (p<0.05). Although the number of non-smoking subjects was low, there was a clear decrease in cloning efficiency observed in keratinocyte cultures for these patients that requires further study. This study found that samples taken from smokers do not produce bystander signals, whereas samples taken from non-smokers can produce such signals following HDR brachytherapy. These findings highlight the importance of studying the interactions of multiple stressors including environmental modifiers with radiation, since some factors such as smoking may elicit protection in tumor cells which could counteract the effectiveness of radiation therapy.

Main radiation pathways in the landscape of Armenia.

PURPOSE: To investigate sources, accumulation, and vertical migration of radionuclides in Armenia, and their impact on biota. CONCLUSIONS: This review describes the radiation status in the landscape of Armenia and features of the impact of natural and human-generated radiation on human and non-human biotas, according to studies of Armenian scientists carried out since the middle of the last century. The mountain landscape demonstrates the diversity, speciation, and radioresistance of the biota, which arise under radiation exposure in a variable environment. Although the effects of radiation have been described for a long time, some of them require further study. It is important to present the data collected in order to produce a base line for future studies of radiation effects and interactions with other stressors caused by climate change.

Use of nail and oral pigmentation to determine ART eligibility among HIV-infected Ugandan adults.

OBJECTIVES: To evaluate the use of grey/distal banded nails as an indicator of advanced immunosuppression, and thus eligibility for ART, in resource poor settings. METHODS: We tested whether grey/distal banded nails and/or oral pigmentation could be used to identify patients with low CD4 cell counts at two cut-offs: <200 and <350 cells/microl in ART naive adults. RESULTS: Four hundred and three nail and oral cavities were photographed and assessed. Grey/distal banded nails and/or oral pigmentation were significantly associated with a CD4 cell count <200 cells/microl (P < 0.001), with a sensitivity of 66%, a specificity of 50% and a negative predictive value of 77%. However, there was no association when a CD4 cell count cut-off of <350 cells/microl was used. Inter-observer agreement (k 0.46) was fair/moderate. CONCLUSIONS: While grey/distal banded nails and/or oral pigmentation are associated with low CD4 counts, the sensitivity and kappa score are too low for this method to be recommended as a tool to guide ART initiation; large number of individuals eligible for ART would be missed.

GSK3beta, a centre-staged kinase in neuropsychiatric disorders, modulates long term memory by inhibitory phosphorylation at serine-9.

Accumulating evidence implicates deregulation of GSK3ss as a converging pathological event in Alzheimer's disease and in neuropsychiatric disorders, including bipolar disorder and schizophrenia. Although these neurological disorders share cognitive dysfunction as a hallmark, the role of GSK3ss in learning and memory remains to be explored in depth. We here report increased phosphorylation of GSK3ss at Serine-9 following cognitive training in two different hippocampus dependent cognitive tasks, i.e. inhibitory avoidance and novel object recognition task. Conversely, transgenic mice expressing the phosphorylation defective mutant GSK3ss[S9A] show impaired memory in these tasks. Furthermore, GSK3ss[S9A] mice displayed impaired hippocampal L-LTP and facilitated LTD. Application of actinomycin, but not anisomycin, mimicked GSK3ss[S9A] induced defects in L-LTP, suggesting that transcriptional activation is affected. This was further supported by decreased expression of the immediate early gene c-Fos, a target gene of CREB. The combined data demonstrate a role for GSK3ss in long term memory formation, by inhibitory phosphorylation at Serine-9. The findings are fundamentally important and relevant in the search for therapeutic strategies in neurological disorders associated with cognitive impairment and deregulated GSK3ss signaling, including AD, bipolar disorder and schizophrenia.

Bystander effect induced changes in apoptosis related proteins and terminal differentiation in in vitro murine bladder cultures.

PURPOSE: Radiation-induced bystander effects are now an established phenomenon seen in numerous cell and tissue culture models. The aim of this investigation was to examine the bystander signal and response in a multicellular primary tissue culture system in vitro. METHODS AND MATERIALS: Murine bladder samples were explanted and directly exposed to gamma radiation, or treated with irradiated tissue conditioned medium (ITCM) generated from the directly irradiated cultures. RESULTS: Results indicated that there was a strong bystander signal produced by the tissue that caused both dose-dependent and -independent changes in the ITCM treated tissue. Significantly increased B-cell lymphoma 2 (Bcl2) expression was noted after treatment with 0.5Gy and 5Gy ITCM (approximately 80%), while dose-dependent changes were observed in c-myelocytomatosis (cMyc) (39.48% at 0.5 Gy ITCM, 81.28% at 5 Gy ITCM) and the terminal differentiation marker uroplakin III (17.88% at 0.5 Gy). Nuclear fragmentation was also significantly increased at both doses of ITCM. CONCLUSION: These data suggest that the bystander signal produced in a multicellular environment induces complex changes in the ITCM-treated culture, and that these changes are reflective of a coordinated response to maintain integrity throughout the tissue.

Intraventricular infusions of anti-NCAM PSA impair the process of consolidation of both avoidance conditioning and spatial learning paradigms in Wistar rats.

Processing of information for long-term storage requires specific patterns of activity that lead to modification of synapse structure and eventual change in neural connectivity pattern. Morphological change associated with memory consolidation is reliant on neural cell adhesion molecule (NCAM) function and that of its polysialylated variant (NCAM PSA). Across species and paradigms, a transient frequency increase of polysialylated neurons in the hippocampal dentate has been found necessary for memory consolidation, however, recent studies suggest that NCAM PSA may serve to suppress memory formation in certain paradigms. As intraventricular infusions of NCAM blocking antibodies have been used successfully to demonstrate its time-dependent role at the 6 h post-training period of memory consolidation, we employed the same procedure to demonstrate a functional requirement for NCAM PSA in the consolidation of two commonly used behavioral paradigms: avoidance conditioning and spatial learning in Wistar rats. Anti-PSA was found to significantly induce amnesia of the passive avoidance response when infused at the 10 h post-training time, a period coincident with the learning-associated increase in dentate polysialylated cell frequency. Moreover, the amnesia became apparent at the 48 h recall time and was not apparent at the 24 h post-training time, suggesting a possible role in memory reconsolidation. A similar anti-PSA action was observed following water maze training in aged animals but was not apparent in young animals, an effect suggested to be due to inadequate antibody saturation of the polysialylated cell population. These studies confirm the requirement for NCAM PSA in memory consolidation and separate it from that of NCAM.

Effect of dose rate on the radiation-induced bystander response.

Radiation-induced biological bystander effects have become a well-established phenomenon associated with the interaction of radiation with cells. These so-called bystander effects have been seen across a variety of end points for both high and low linear energy transfer (LET) radiations, utilizing a variety of dose rates and radiation sources. In this study, the effect of dose rate and different low LET sources on the bystander cell survival fraction (SF) was examined. The cell line investigated was the human keratinocyte HPV-G. The bystander response was measured via clonogenic assay after medium transfer protocol. Cells were irradiated using (60)Co gamma-rays and 20 MeV electrons at doses of 0.5, 5 and 10 Gy with varying dose rates. Both gamma and electron irradiation decreased recipient SF at 0.5 Gy and 5 Gy, respectively. Subsequent recovery of the SF to control levels for 10 Gy was observed. There was no dose rate dependence for (60)Co irradiation. A significant difference in the survival fraction was observed for electron irradiation at 10 Gy and a high dose rate. Furthermore, survival fractions were compared between (60)Co and 20 MeV electron irradiations. This showed a significant increase in the survival fraction 'recovery' at 10 Gy for a (60)Co dose rate of 1.1 Gy min(-1) compared to 20 MeV electrons at 1.0 Gy min(-1). No such difference was observed when comparing at higher dose rates. Lastly, increases in survival fraction at 10 Gy were abolished and the SF decreased by the plating of increased numbers of recipient cells. Such evidence may help gain insight into the nature and mechanism(s) surrounding bystander signal production, how these phenomena are tested and their eventual application in a clinical setting.

Point-of-care sensors for the management of sepsis.

Point-of-care sensors that enable the fast collection of information relevant to a patient's health state can facilitate improved health access, reduce healthcare costs and improve the quality of healthcare delivery. In the diagnosis of sepsis - defined as a life-threatening organ dysfunction caused by a dysregulated host response to infection, and the leading cause of in-patient death and of hospital readmission in the United States - predicting which infections will lead to life-threatening organ dysfunction and developing specific anti-sepsis treatments remain challenging because of the significant heterogeneity of the host response. Yet the use of point-of-care devices could reduce the time from the onset of a patient's infection to the administration of appropriate therapeutics. In this Perspective, we describe the current state of point-of-care sensors for the diagnosis and monitoring of sepsis, and outline opportunities in the use of these devices to dramatically improve patient care.

Multiple stressor effects of radiation and metals in salmon (Salmo salar).

These experiments were designed to look at the cellular effects in key organs in Atlantic salmon (Salmo salar) after exposure in vivo to radiation and subtoxic levels of aluminum (Al) and cadmium (Cd), alone or in combination. Salmon (25g) were exposed to a single 0.5Gy dose of gamma-irradiation in water containing Cd, Al or Cd+Al. Three fish per group were sacrificed after 1h and the liver, pronephros, fin and gill of each was dissected. Small explants of each tissue were set up. After 2 days, the culture medium was harvested and filtered then placed on a reporter cell line for determination of stress signal activity (bystander effects). Radiation in combination with Cd and/or Al, caused bystander effects in tissues harvested from in vivo exposed salmon. The effects vary between different organs and are not consistently additive or synergistic for a given treatment. Tissue type appears to be critical. Liver cultures produce a toxic factor which is lethal to reporter cells, and therefore no liver data could be obtained. It is hoped that this stress signal response will prove to be a useful indicator of environmental stress in species inhabiting aquatic ecosystems.

An Observed Effect of p53 Status on the Bystander Response to Radiation-Induced Cellular Photon Emission.

In this study, we investigated the potential influence of p53 on ultraviolet (UV) signal generation and response of bystander cells to the UV signals generated by beta-irradiated cells. Five cell lines of various p53 status (HaCaT, mutated; SW48, wild-type; HT29, mutated; HCT116+/+, wild-type; HCT116-/-, null) were irradiated with beta particles from tritium. Signal generation (photon emission at 340 ± 5 nm) was quantified from irradiated cells using a photomultiplier tube. Bystander response (clonogenic survival) was assessed by placing reporter cell flasks directly superior to irradiated signal-emitting cells. All cell lines emitted significant quantities of UV after tritium exposure. The magnitudes of HaCaT and HT29 photon emission at 340 nm were similar to each other while they were significantly different from the stronger signals emitted from SW48, HCT116+/+ and HCT116-/- cells. In regard to the bystander responses, HaCaT, HCT116+/+ and SW48 cells demonstrated significant reductions in survival as a result of exposure to emission signals. HCT116-/- and HT29 cells did not exhibit any changes in survival and thus were considered to be lacking the mechanisms or functions required to elicit a response. The survival response was found not to correlate with the observed signal strength for all experimental permutations; this may be attributed to varying emission spectra from cell line to cell line or differences in response sensitivity. Overall, these results suggest that the UV-mediated bystander response is influenced by the p53 status of the cell line. Wild-type p53 cells (HCT116+/+ and SW48) demonstrated significant responses to UV signals whereas the p53-null cell line (HCT116-/-) lacked any response. The two mutated p53 cell lines exhibited contrasting responses, which may be explained by unique modulation of functions by different point mutations. The reduced response (cell death) exhibited by p53-mutated cells compared to p53 wild-type cells suggests a possible role of the assessed p53 mutations in radiation-induced cancer susceptibility and reduced efficacy of radiation-directed therapy.

Actions of radiation on living cells in the "post-bystander" era.

Over the past 20 years there has been increasing evidence that cells and the progeny of cells surviving a dose of ionizing radiation can exhibit a wide range of effects inconsistent with the level of dose received. Recently, the cause of these delayed effects has been ascribed to so-called bystander effects, occurring in cells not directly hit by an ionizing track, but which are influenced by signals from irradiated cells. These effects are not necessarily deleterious, although most of the literature deals with adverse delayed effects. What is important to consider is what, if anything, these effects mean for what is still the central dogma of radiobiology and radiation protection, i.e., that DNA double-strand breaks are the primary radiation-induced lesion that can be quantifiably related to received dose, and which determine the probability that a cancer will result from a radiation exposure. In this chapter we review the history of radiation biology which led to the DNA paradigm. We explore the issues and the evidence which are now challenging the view that dose deposition in DNA is all important. We conclude that in the low-dose region, the primary determinant of radiation exposure outcome is the genetic and epigenetic background of the individual and not the dose. This effectively dissociates dose from effect as a quantitative relationship, but it does not necessarily mean that the effect is unrelated to DNA damage somewhere in the system.

Increased radiosensitivity in cells of two human cell lines treated with bystander medium from irradiated repair-deficient cells.

Radiation-induced bystander factors have been shown to be more toxic if they are from medium harvested from irradiated repair-deficient cells. The aim of this study was to test the hypothesis that the radiosensitivity of repair-proficient cells can be increased by exposing them to medium-borne factors harvested from sensitive cells and vice versa. Cells from a mismatch repair (MMR)-deficient cell line (Raji 10) with a sensitive response to radiation or the wild-type parent cell line were irradiated to 0.5 Gy gamma rays and then monitored for growth rate in their own medium or in the alternative conditioned medium. In other experiments, cells or conditioned medium were added to reporter cells (HPV-G, which are relatively sensitive keratinocytes, or highly radioresistant HT29 cells). The subsequent responses of the two cell lines to a 0.5-Gy dose of (60)Co gamma rays were measured. The results show that prior exposure of resistant cells to medium from irradiated sensitive cells reduced the clonogenic survival of the subsequently irradiated resistant cells. The reverse is also true. Measurement of the apoptosis index and BCL2 expression confirmed that the harvested medium was capable of modulating apoptosis after irradiation. This may have important applications in tumor therapy and also in the understanding of mechanisms involved in induction of adaptive responses.

The involvement of calcium and MAP kinase signaling pathways in the production of radiation-induced bystander effects.

Much evidence now exists regarding radiation-induced bystander effects, but the mechanisms involved in the transduction of the signal are still unclear. The mitogen-activated protein kinase (MAPK) pathways have been linked to growth factor-mediated regulation of cellular events such as proliferation, senescence, differentiation and apoptosis. Activation of multiple MAPK pathways such as the ERK, JNK and p38 pathways have been shown to occur after exposure of cells to radiation and a variety of other toxic stresses. Previous studies have shown oxidative stress and calcium signaling to be important in radiation-induced bystander effects. The aim of the present study was to investigate MAPK signaling pathways in bystander cells exposed to irradiated cell conditioned medium (ICCM) and the role of oxidative metabolism and calcium signaling in the induction of bystander responses. Human keratinocytes (HPV-G cell line) were irradiated (0.005-5 Gy) using a cobalt-60 teletherapy unit. The medium was harvested 1 h postirradiation and transferred to recipient HPV-G cells. Phosphorylated forms of p38, JNK and ERK were studied by immunofluorescence 30 min-24 h after exposure to ICCM. Inhibitors of the ERK pathway (PD98059 and U0126), the JNK pathway (SP600125), and the p38 pathway (SB203580) were used to investigate whether bystander-induced cell death could be blocked. Cells were also incubated with ICCM in the presence of superoxide dismutase, catalase, EGTA, verapamil, nifedipine and thapsigargin to investigate whether bystander effects could be inhibited because of the known effects on calcium homeostasis. Activated forms of JNK and ERK proteins were observed after exposure to ICCM. Inhibition of the ERK pathway appeared to increase bystander-induced apoptosis, while inhibition of the JNK pathway appeared to decrease apoptosis. In addition, reactive oxygen species, such as superoxide and hydrogen peroxide, and calcium signaling were found to be important modulators of bystander responses. Further investigations of these signaling pathways may aid in the identification of novel therapeutic targets.

Radiation-induced bystander effects and the DNA paradigm: an "out of field" perspective.

Over the past 20 years there has been increasing evidence that cells and the progeny of cells surviving a very low dose of ionizing radiation [micro-mGy] can exhibit a wide range of non-monotonic effects such as adaptive responses, low dose hypersensitivity and other delayed effects. These effects are inconsistent with the expected dose-response, when based on extrapolation of high dose data and cast doubt on the reliability of extrapolating from high dose data to predict low dose effects. Recently the cause of many of these effects has been tentatively ascribed to so-called "bystander effects". These are effects that occur in cells not directly hit by an ionizing track but which are influenced by signals from irradiated cells and are thus highly relevant in situations where the dose is very low. Not all bystander effects may be deleterious although most endpoints measured involve cell damage or death. In this commentary, we consider how these effects impact the historical central dogma of radiobiology and radiation protection, which is that DNA double strand breaks are the primary radiation-induced lesion which can be quantifiably related to received dose and which determine the probability that a cancer will result from a radiation exposure. We explore the low dose issues and the evidence and conclude that in the very low dose region, the primary determinant of radiation exposure outcome is the genetic and epigenetic background of the individual and not solely the dose. What this does is to dissociate dose from effect as a quantitative relationship, but it does not necessarily mean that the effect is ultimately unrelated to DNA damage. The fundamental thesis we present is that at low doses fundamentally different mechanisms underlie radiation action and that at these doses, effect is not quantitatively related to dose.

The effect of melanin on the bystander effect in human keratinocytes.

The influence of melanin on radiation-induced bystander effects has been studied. Melanin is known to be a natural substance with proved radioprotective properties in different organisms and cell lines. It is non-toxic and is effective against acute and chronic irradiation. The lower the radiation dose, the higher the relative impact of melanin protection. In this study influence of melanin on human keratinocytes (HPV-G cells) has been studied using the colony-forming assay. We have shown that bystander donor medium from 0.5 Gy irradiated cells when transferred to unirradiated cells, caused almost the same effect as direct irradiation. Melanin increased the colony-forming ability of bystander recipient cells when it was added into culture medium before irradiation. The effect of melanin added after irradiation was to produce less protection in both the directly irradiated and bystander medium treated groups. The absorption spectrum of the filtered medium is identical to one of the intact culture medium showing that melanin was not present in filtered medium. Thus, it cannot protect recipient cells but reduces the amount of the bystander effect. It is concluded that melanin added before irradiation effectively decreased the radiation dose. The reduction of the impact of the bystander signal on recipient cells when melanin was added to the donor medium after harvest but before filtration, may mean that the bystander signal has a physical component as melanin can absorb all types of physical energy.

Effect of low inspired oxygen fraction on respiratory indices in mechanically ventilated horses anaesthetised with isoflurane and medetomidine constant rate infusion.

Horses may become hypoxaemic during anaesthesia despite a high inspired oxygen fraction (FiO2). A lower FiO2 is used commonly in human beings to minimise atelectasis and to improve lung function, and previously has been shown to be of potential benefit in horses in experimental conditions. Other studies suggest no benefit to using a FiO2 of 0.5 during clinically relevant conditions; however, low FiO2 (0.65) is commonly used in practice and in a large number of studies. The present study was performed to compare the effect of a commonly used FiO2 of 0.65 versus 0.90 on calculated respiratory indices in anaesthetised mechanically ventilated horses in a clinical setting. Eighteen healthy Thoroughbred horses anaesthetised for experimental laryngeal surgery were recruited into a prospective, non-blinded, randomised clinical study. Before anaesthesia, the horses were randomly allocated into either low (0.65) or high (0.90) FiO2 groups and arterial blood gas (ABG) analysis was performed every 30 min during anaesthesia to allow for statistical analysis of respiratory indices. As expected, PaO2 was significantly lower in horses anaesthetised with a low FiO2, but was sufficient to fully saturate haemoglobin. There were no significant improvements in any of the other respiratory indices. There is no obvious benefit to be gained from the use of a FiO2 of 0.65 compared to 0.90 for mechanically ventilated Thoroughbred horses anaesthetised in lateral recumbency with isoflurane and a medetomidine constant rate infusion.

The nature of the Ca2+-pump defect in the red blood cells of patients with cystic fibrosis.

The reduction in (Ca2+ + Mg2+)-ATPase activity in the cystic fibrosis red blood cells can be attributed to a reduction in the number of active Ca2+ pumps per red blood cell and an altered interaction of calcium ions with the pump. Despite this, the normal free intracellular [Ca2+] is preserved due to a lower rate of passive calcium entry.