Non-targeted Effects Research Articles

Linking Microdosimetric Measurements to Biological Effectiveness in Ion Beam Therapy: A Review of Theoretical Aspects of MKM and Other Models

Different qualities of radiation are known to cause different biological effects at the same absorbed dose. Enhancements of the biological effectiveness are a direct consequence of the energy deposition clustering at the scales of DNA molecule and cell nucleus whilst absorbed dose is a macroscopic averaged quantity which does not take into account heterogeneities at the nanometer and micrometer scales. Microdosimetry aims to measure radiation quality at cellular or sub-cellular levels trying to increase the understanding of radiation damage mechanisms and effects. Existing microdosimeters rely on the well-established gas-based detectors or the more recent solid-state devices. They provide specific energy z spectra and other derived quantities as lineal energy (y) spectra assessed at the micrometer level. The interpretation of the radio-biological experimental data in the framework of different models has raised interest and various investigations have been performed to link in vitro and in vivo radiobiological outcomes with the observed microdosimetric data. A review of the major models based on experimental microdosimetry, with a particular focus on ion beam therapy applications and an emphasis on the microdosimetric kinetic model (MKM), will be presented in this work, enlightening the advantages of each one in terms of accuracy, initial assumptions, and agreement with experimental data. The MKM has been used to predict different kinds of radiobiological quantities such as the relative biological effects for cell inactivation or the oxygen enhancement ratio. Recent developments of the MKM will be also presented, including new non-Poissonian correction approaches for high linear energy transfer radiation, the inclusion of partial repair effects for fractionation studies, and the extension of the model to account for non-targeted effects. We will also explore developments for improving the models by including track structure and the spatial damage correlation information, by using the full fluence spectrum and by better accounting for the energy-deposition fluctuations at the intra- and inter-cellular level.

ATP-triggered drug release system based on ZIF-90 loaded porous poly(lactic-co-glycolic acid) microspheres

Poly(lactic-co-glycolic acid) (PLGA) based microparticles have been widely applied in drug delivery system, but pristine PLGA particles suffers from low efficiency in release and non-targeted effect for drug delivery system. Inspired by cancer cells environment, we have fabricated an ATP-triggered delivery system based on porous PLGA microspheres covering drug-loaded ZIF-90. The system contains controlled doxorubicin releasing because ZIF-90 nanocrystals are easily collapsed under exposing to high-concentration ATP environment. In addition, ZIF-90@PLGA microparticles also perform low toxicity and 2.37 % hemolysis, and improved fluorescence intensity of doxorubicin by loading riboflavin sodium phosphate benefited from their partial Förster resonance energy transfer. This novel ATP-triggered drug delivery system based on porous PLGA particles could be widely utilized to design effective drug release biomaterials for gastric and lung cancer.

DNA-Induced Restructuring of Interfacial Water and the Hydrocarbon Chain of Different Forms of Fungicide at the Water Interface: Vibrational Sum Frequency Generation Study

Fungicides adopt different structures at the air/water interface depending on their concentration, which affects their potency against fungi. However, the nontargeted action of fungicides causes a genotoxic effect on other organisms, which is responsible for several serious diseases such as cancer, neurodegenerative disorders, and developmental problems. Hence, understanding the effect of DNA on the different structures of fungicides at the air/water interface is essential. The surface-selective vibrational sum frequency generation technique has been implemented to obtain insights into the effect of DNA on the fungicide, namely, dodine at the air/water interface. The hydrocarbon chain of dodine with an increasing concentration adopts disordered, ordered, and packed forms at the air/water interface. DNA gets closer to the positively charged head group of dodine at the air/water interface in its disordered, ordered, and packed forms by removing the interfacial water and reorganizes the hydrogen bonding of water beneath the head group of dodine. Interestingly, DNA also increases the ordering of the hydrocarbon chain of disordered and ordered dodine at the air/water interface by modulating the gauche defect and orientational ordering of the hydrocarbon chain of dodine. Unlike the disordered and ordered dodine, the addition of DNA at the packed dodine on the air/water interface does not affect the ordering of its hydrocarbon chain. The significant reorganization of the interfacial water and the hydrocarbon chain of dodine by DNA can be useful for understanding the mechanism of the nontargeted genotoxic effects of dodine.

Isolation of the effects of alpha-related components from total effects of radium at low doses

Purpose Radium is the most common source of alpha radiation exposure to humans and non-human species in the environment but the dosimetry is complicated by the decay chain which involves gamma exposure due to radon daughters. This paper seeks to determine the separate contributions of alpha and gamma doses to the total dose and total direct and non-targeted effect in a fish and a human cell line. Materials and methods This study aimed to isolate the effect of alpha particles following exposure to low doses of radium in cells, and their progeny which received no further exposure. This was initially done by comparing the survival values of a human keratinocyte cell line (HaCaT) and an embryonic Chinook salmon cell line (CHSE-214) exposed to gamma radiation, from survival of the same cell lines exposed to mixed alpha and gamma radiation through exposure to Ra-226 and its decay products. A Monte Carlo simulation was later performed to determine the contributions of radium decay products including radon daughters. Results The human cell line showed increased radioresistance when exposed to low doses of alpha particles. In contrast the fish cell line, which demonstrated radioresistance to low dose gamma radiation, showed increased lethality when exposed to low doses of alpha particles. Significant and complex levels of non-targeted effects were induced in progeny of irradiated cells. The simulation showed that gamma and beta decay products did not contribute significant dose and the highest beta dose was below the threshold for inducing non-targeted effects. Conclusions The results confirm the need to consider the dose-response relationship when developing radiation weighting factors for low dose exposures, as well as the need to be aware of possible cell line and species differences.

Transcriptional modulations induced by proton irradiation in mice skin in function of adsorbed dose and distance

ABSTRACT Hadron therapy by proton beams represents an advanced anti-cancer strategy due to its highly localized dose deposition allowing a greater sparing of normal tissue and/or organs at risk compared to photon/electron radiotherapy. However, it is not clear to what extent non-targeted effects such as transcriptional modulations produced along the beamline may diffuse and impact the surrounding tissue. In this work, we analyze the transcriptome of proton-irradiated mouse skin and choose two biomarker genes to trace their modulation at different distances from the beam’s target and at different doses and times from irradiation to understand to what extent and how far it may propagate, using RNA-Seq and quantitative RT-PCR. In parallel, assessment of lipids alteration is performed by FTIR spectroscopy as a measure of tissue damage. Despite the observed high individual variability of expression, we can show evidence of transcriptional modulation of two biomarker genes at considerable distance from the beam’s target where a simulation system predicts a significantly lower adsorbed dose. The results are compatible with a model involving diffusion of transcripts or regulatory molecules from high dose irradiated cells to distant tissue’s portions adsorbing a much lower fraction of radiation.

REVIEW OF QUANTITATIVE MECHANISTIC MODELS OF RADIATION-INDUCED NON-TARGETED EFFECTS (NTE).

Quantitative mechanistic modeling of the biological effects of ionizing radiation has a long rich history. Initially, it was dominated by target theory, which quantifies damage caused by traversal of cellular targets like DNA by ionizing tracks. The discovery that mutagenesis, death and/or altered behavior sometimes occur in cells that were not themselves traversed by any radiation tracks but merely interacted with traversed cells was initially seen as surprising. As more evidence of such 'non-targeted' or 'bystander' effects accumulated, the importance of their contribution to radiation-induced damage became more recognized. Understanding and modeling these processes is important for quantifying and predicting radiation-induced health risks. Here we review the variety of mechanistic mathematical models of nontargeted effects that emerged over the past 2-3 decades. This review is not intended to be exhaustive, but focuses on the main assumptions and approaches shared or distinct between models, and on identifying areas for future research.

The Response of Living Organisms to Low Radiation Environment and Its Implications in Radiation Protection

Life has evolved on Earth for about 4 billion years in the presence of the natural background of ionizing radiation. It is extremely likely that it contributed, and still contributes, to shaping present form of life. Today the natural background radiation is extremely small (few mSv/y), however it may be significant enough for living organisms to respond to it, perhaps keeping memory of this exposure. A better understanding of this response is relevant not only for improving our knowledge on life evolution, but also for assessing the robustness of the present radiation protection system at low doses, such as those typically encountered in everyday life. Given the large uncertainties in epidemiological data below 100 mSv, quantitative evaluation of these health risk is currently obtained with the aid of radiobiological models. These predict a health detriment, caused by radiation-induced genetic mutations, linearly related to the dose. However a number of studies challenged this paradigm by demonstrating the occurrence of non-linear responses at low doses, and of radioinduced epigenetic effects, i.e., heritable changes in genes expression not related to changes in DNA sequence. This review is focused on the role that epigenetic mechanisms, besides the genetic ones, can have in the responses to low dose and protracted exposures, particularly to natural background radiation. Many lines of evidence show that epigenetic modifications are involved in non-linear responses relevant to low doses, such as non-targeted effects and adaptive response, and that genetic and epigenetic effects share, in part, a common origin: the reactive oxygen species generated by ionizing radiation. Cell response to low doses of ionizing radiation appears more complex than that assumed for radiation protection purposes and that it is not always detrimental. Experiments conducted in underground laboratories with very low background radiation have even suggested positive effects of this background. Studying the changes occurring in various living organisms at reduced radiation background, besides giving information on the life evolution, have opened a new avenue to answer whether low doses are detrimental or beneficial, and to understand the relevance of radiobiological results to radiation protection.

Some Aspects Related to Transmission of Radiation-Induced Alterations due to the Bystander Effect

This paper presents the first results of research and observations within the framework of the Integrated Project for the assessment of radiation-induced nontargeted effects. The remote bystander effect, related to nontargeted effects, is one of the least studied phenomena in terms of manifestations and mechanisms. The international laboratory practice with respect to this phenomenon commenced with demonstration of the bystander effect in fish and other aquacultures. Our studies have revealed changes similar to radiation-induced pathologies in nonirradiated mice kept for three months in contact with the irradiated species (exposed to a 3 Gy γ-ray dose). Changes in behavioral responses, a statistically significant trend for decreasing spleen weight (r = –0.416; p = 0.048), an expanding area of alopecia (r = –0.631; p = 0.001), and other anomalies are noted. Discussions on possible mechanisms of the phenomena refer to both A.M. Kuzin’s theory of radiotoxins (1966) and the interpretation of the hypotheses investigated by C. Mothersill (2010–2019). An assumption is made on the important role of derivatives of lipid peroxides in the formation of this effect. The results of these studies indicate the need for further investigation of the radiation-induced bystander effect by using the molecular and cellular methodological approaches.

Dynamics of morphological changes in neural cell culture with a model of neurotrauma under the influence of conditioned media of the rat fetal brain neurogenic cells

A potential strategy for recovery and regeneration of brain damage due to traumatic brain injury is considered to be the transplantation of neurogenic stem and/or progenitor cells (NSCs/NPCs). The key factors of the regenerative non-targeted effects of NSCs/NPCs (so-called bystander effects) include the signal molecules produced by them into the extracellular environment (secretome). The purpose is to study the regenerative bystander effects of rat fetal brain neurogenic cells (FBNCs) in the in vitro model of neurotrauma. Materials and methods. In cell culture of FBNCs from rat fetuses (E14-16), neurotrauma was modeled in vitro by mechanical scratching of monolayer and conditioned medium obtained from 24-h cultures of rat FBNCs was added. Cell phenotype was evaluated by morphological features and by immunocytochemical staining for Nestin and GFAP. The density and length of processes, migration capacity, the cell growth rate and monolayer density in the scratched area were compared. Morphometric study included analysis of the width of the scratched area, the number of migrating cells, the distance of migration and mitotic activity in the intact monolayer. Results. Under the conditions of the nutrient medium of standard composition in the scratched area the signs of endogenous regeneration are shown during 24-48 h of cultivation. The overgrowth of cell processes from monolayer and short distance migration of single undifferentiated or poorly differentiated cells were shown. In the next 72-96 h of observation, the degeneration of migrated cells and processes in the scratched area was detected. Under the influence of conditioned media from 24-h cultures of FBNCs by single addition immediately after scratching at dose of 0.1 mg/ml for protein content the stimulation of regeneration were detected up to 96 hours of cultivation. The migration of cell processes from the monolayer simultaneously with undifferentiated or poorly differentiated cells at 24 hours was shown. The formation of cell clusters and their differentiation (at 48 h), as well as migration of differentiated cells with partial or complete overgrowth of scratched area (72-96 h) were observed. The morphological signs of degeneration of migrated cells in the scratched area appeared only on the 8th day of cultivation. Conditioned media does not affect qualitative and quantitative properties of the culture of rat FBNCs in the intact area where mitotic activity was average. Conclusions. Conditioned medium from 24-h cultures of rat FBNC can stimulate reparation in the in vitro model of neurotrauma in neural cell culture for at least 7 days at a single addition, without affecting the cellular composition and mitotic activity of the intact monolayer.

Phenotypic and Functional Characteristics of Exosomes Derived from Irradiated Mouse Organs and Their Role in the Mechanisms Driving Non-Targeted Effects.

Molecular communication between irradiated and unirradiated neighbouring cells initiates radiation-induced bystander effects (RIBE) and out-of-field (abscopal) effects which are both an example of the non-targeted effects (NTE) of ionising radiation (IR). Exosomes are small membrane vesicles of endosomal origin and newly identified mediators of NTE. Although exosome-mediated changes are well documented in radiation therapy and oncology, there is a lack of knowledge regarding the role of exosomes derived from inside and outside the radiation field in the early and delayed induction of NTE following IR. Therefore, here we investigated the changes in exosome profile and the role of exosomes as possible molecular signalling mediators of radiation damage. Exosomes derived from organs of whole body irradiated (WBI) or partial body irradiated (PBI) mice after 24 h and 15 days post-irradiation were transferred to recipient mouse embryonic fibroblast (MEF) cells and changes in cellular viability, DNA damage and calcium, reactive oxygen species and nitric oxide signalling were evaluated compared to that of MEF cells treated with exosomes derived from unirradiated mice. Taken together, our results show that whole and partial-body irradiation increases the number of exosomes, instigating changes in exosome-treated MEF cells, depending on the source organ and time after exposure.

Characterization of exosome release and extracellular vesicle-associated miRNAs for human bronchial epithelial cells irradiated with high charge and energy ions

Exosomes are extracellular vesicles that mediate transport of nucleic acids, proteins, and other molecules. Prior work has implicated exosomes in the transmission of radiation nontargeted effects. Here we investigate the ability of energetic heavy ions, representative of species found in galactic cosmic rays, to stimulate exosome release from human bronchial epithelial cells in vitro. Immortalized human bronchial epithelial cells (HBEC3-KT F25F) were irradiated with 1.0Gy of high linear energy transfer (LET) 48Ti, 28Si, or 16O ions, or with 10Gy of low-LET reference γ-rays, and extracellular vesicles were collected from conditioned media. Preparations were characterized by single particle tracking analysis, transmission electron microscopy, and immunoblotting for the exosomal marker, TSG101. Based on TSG101 levels, irradiation with high-LET ions, but not γ-rays, stimulated exosome release by about 4-fold, relative to mock-irradiated controls. The exosome-enriched vesicle preparations contained pro-inflammatory damage-associated molecular patterns, including HSP70 and calreticulin. Additionally, miRNA profiling was performed for vesicular RNAs using NanoString technology. The miRNA profile was skewed toward a small number of species that have previously been shown to be involved in cancer initiation and progression, including miR-1246, miR-1290, miR-23a, and miR-205. Additionally, a set of 24 miRNAs was defined as modestly over-represented in preparations from HZE ion-irradiated versus other cells. Gene set enrichment analysis based on the over-represented miRNAs showed highly significant association with nonsmall cell lung and other cancers.

Exosomes and exosomal microRNA in non-targeted radiation bystander and abscopal effects in the central nervous system.

Localized cranial radiotherapy is a dominant treatment for brain cancers. After being subjected to radiation, the central nervous system (CNS) exhibits targeted effects as well as non-targeted radiation bystander effects (RIBE) and abscopal effects (RIAE). Radiation-induced targeted effects in the CNS include autophagy and various changes in tumor cells due to radiation sensitivity, which can be regulated by microRNAs. Non-targeted radiation effects are mainly induced by gap junctional communication between cells, exosomes containing microRNAs can be transduced by intracellular endocytosis to regulate RIBE and RIAE. In this review, we discuss the involvement of microRNAs in radiation-induced targeted effects, as well as exosomes and/or exosomal microRNAs in non-targeted radiation effects in the CNS. As a target pathway, we also discuss the Akt pathway which is regulated by microRNAs, exosomes, and/or exosomal microRNAs in radiation-induced targeted effects and RIBE in CNS tumor cells. As the CNS-derived exosomes can cross the blood-brain-barrier (BBB) into the bloodstream and be isolated from peripheral blood, exosomes and exosomal microRNAs can emerge as promising minimally invasive biomarkers and therapeutic targets for radiation-induced targeted and non-targeted effects in the CNS.

Irradiation of the kidneys causes pathologic remodeling in the nontargeted heart: A role for the immune system.

ABSTRACTCardiac disease is a frequent and significant adverse event associated with radiotherapy for cancer. Identifying the underlying mechanism responsible for radiation injury to the heart will allow interventions to be developed. In the present study, we tested if local kidney irradiation results in remodeling of the shielded, nontargeted heart. One kidney, two kidneys, or the total body of male WAG and Dahl SS rats were irradiated with 10 Gy of X‐rays. Local kidney irradiation resulted in systemic hypertension, increased BUN, infiltration of T lymphocytes, natural killer cells, and macrophages into the renal cortex and medulla, and renal fibrosis. Local irradiation of kidneys in WAG rats resulted in remodeling in the nontargeted heart after 120 days, manifested by perivascular fibrosis and increased interventricular septal thickness, but was not seen in Dahl SS rats due to a high baseline level of fibrosis in the sham‐irradiated animals. Genetic depletion of T cells mitigated the nephropathy after local kidney irradiation, indicating a role for the immune system in mediating this outcome. Local kidney irradiation resulted in a cascade of pro‐inflammatory cytokines and low‐molecular weight metabolites into the circulation associated with transmission of signals resulting in pathologic remodeling in the nontargeted heart. A new model is proposed whereby radiation‐induced cardiac remodeling in susceptible animals is indirect, with lower hemi body organs such as the kidney exporting factors into the circulation that cause remodeling outside of the irradiated field in the shielded, nontargeted heart. This nontargeted effect appears to be mediated, in part, by the immune system.

Inoculation With the Plant-Growth-Promoting Rhizobacterium Pseudomonas fluorescens LBUM677 Impacts the Rhizosphere Microbiome of Three Oilseed Crops.

The bacterial communities inhabiting the rhizosphere play an important role in plant development and health. Here we studied the effect of inoculation with Pseudomonas fluorescens LBUM677, a plant growth promoting rhizobacterium that promotes seed oil accumulation, on the rhizosphere microbiome of three oilseed crops (Brassica napus, Buglossoides arvensis, and Glycine max) over time. Next-Generation high-throughput sequencing targeting the V4 region of 16S rDNA was used to characterize the microbial communities associated with the three different crops, inoculated or not with LBUM677, over a time period of up to 90 days post-inoculation. A total of 1,627,231 amplicon sequence variants were obtained and were taxonomically grouped into 39 different phyla. LBUM677 inoculation and sampling date were found to significantly influence the rhizosphere microbiome of the three oil-producing crops under study. Specifically, inoculation with LBUM677 and sampling date, but not the plant species, were found to significantly alter the alpha- and the beta-diversity of the rhizosphere microbial communities. Differential abundance analyses found that 29 taxonomical bacterial groups were significantly more abundant in the LBUM677 treatments while 30 were significantly more abundant in the control treatments. Predicted functions of the microorganisms were also enriched, including 47 enzymatic pathways in LBUM677 treatments. These non-targeted effects on rhizosphere bacterial communities are discussed in the context of oilseed crops.

Radiation Effects on Brain Extracellular Matrix.

Radiotherapy is an important therapeutic approach to treating malignant tumors of different localization, including brain cancer. Glioblastoma multiforme (GBM) represents the most aggressive brain tumor, which develops relapsed disease during the 1st year after the surgical removal of the primary node, in spite of active adjuvant radiochemotherapy. More and more evidence suggests that the treatment's success might be determined by the balance of expected antitumor effects of the treatment and its non-targeted side effects on the surrounding brain tissue. Radiation-induced damage of the GBM microenvironment might create tumor-susceptible niche facilitating proliferation and invasion of the residual glioma cells and the disease relapse. Understanding of molecular mechanisms of radiation-induced changes in brain ECM might help to reconsider and improve conventional anti-glioblastoma radiotherapy, taking into account the balance between its antitumor and ECM-destructing activities. Although little is currently known about the radiation-induced changes in brain ECM, this review summarizes current knowledge about irradiation effects onto the main components of brain ECM such as proteoglycans, glycosaminoglycans, glycoproteins, and the enzymes responsible for their modification and degradation.

Ceramide-Enriched Membrane Domains Contribute to Targeted and Nontargeted Effects of Radiation through Modulation of PI3K/AKT Signaling in HNSCC Cells.

We investigated the potential involvement of ceramide-enriched membrane domains in radiation-induced targeted and nontargeted effects using head and neck squamous cell carcinoma with opposite radiosensitivities. In radiosensitive SCC61 cells, the proportion of targeted effects was 34% and nontargeted effects killed 32% of cells. In contrast, only targeted effects (30%) are involved in the overall death of radioresistant SQ20B cells. We then demonstrated in SCC61 cells that nontargeted cell response was driven by the formation of the radiation-induced ceramide-enriched domain. By contrast, the existence of these platforms in SQ20B cells confers a permissive region for phosphatidylinositol-3-kinase (PI3K)/AKT activation. The disruption of lipid raft results in strong inhibition of PI3K/AKT signaling, leading to radiosensitization and apparition of nontargeted effects. These results suggest that ceramide-enriched platforms play a significant role in targeted and nontargeted effects during radiotherapy and that drugs modulating cholesterol levels may be a good alternative for improving radiotherapy effectiveness.

Database of Families of Workers Chronically Exposed to Radiation: Data and Biospecimen Resources.

Animal experiment findings suggest that high doses of ionizing radiation exposure (>1.0 Gy) may cause genetic and epigenetic effects in offspring. However, epidemiological studies of offspring of radiation-exposed parents did not find increased risks of any health effects. Findings of cellular/experimental investigations and studies of human health effects are contradicting, and further investigations are needed to help resolve ambiguities using updated and/or improved data. This paper provides a detailed description of a database of families of workers of the first Russian nuclear facility, Mayak Production Association, located in the Southern Urals in the Chelyabinsk region close to Ozyorsk city, which started its operation in 1948 and today consists of reactors, radiochemical and plutonium production plants, and auxiliary facilities. The Mayak worker cohort includes 22,377 individuals (25% females) who were hired at one of the main Mayak PA facilities between 1948 and 1982 and were externally or internally exposed to ionizing radiation over prolonged periods. Advantages of the cohort include its large size, extensive follow-up period (70 y), individually measured doses from external and internal exposure and the wide range of these doses, heterogeneity by gender/age/ethnicity/initial health status, complete data on vital status and causes of death, available medical information on morbidity and reproduction, available data on non-radiation factors, and stored biological specimens donated by more than one-third of the cohort members. Based on medical and dosimetry database "Clinics" containing raw data on workers of the study cohort, the Mayak workers' family and offspring database was created. To date, it comprises 12,195 family couples (a husband and a wife) and 16,585 offspring. Biological specimens are available for more than 1,000 family triads (a husband, a wife, and their child). Stages of assembling the database and its descriptive characteristics are presented in this paper. Examples of potential applications of the database for investigations of non-targeted and transgenerational radiation effects in offspring of exposed parents are discussed.

Benchmarking risk predictions and uncertainties in the NSCR model of GCR cancer risks with revised low let risk coefficients

We report on the contributions of model factors that appear in projection models to the overall uncertainty in cancer risks predictions for exposures to galactic cosmic ray (GCR) in deep space, including comparisons with revised low LET risks coefficients. Annual GCR exposures to astronauts at solar minimum are considered. Uncertainties in low LET risk coefficients, dose and dose-rate modifiers, quality factors (QFs), space radiation organ doses, non-targeted effects (NTE) and increased tumor lethality at high LET compared to low LET radiation are considered. For the low LET reference radiation parameters we use a revised assessment of excess relative risk (ERR) and excess additive risk (EAR) for radiation induced cancers in the Life-Span Study (LSS) of the Atomic bomb survivors that was recently reported, and also consider ERR estimates for males from the International Study of Nuclear Workers (INWORKS). For 45-y old females at mission age the risk of exposure induced death (REID) per year and 95% confidence intervals is predicted as 1.6% [0.71, 1.63] without QF uncertainties and 1.64% [0.69, 4.06] with QF uncertainties. However, fatal risk predictions increase to 5.83% [2.56, 9.7] based on a sensitivity study of the inclusion of non-targeted effects on risk predictions. For males a comparison using LSS or INWORKS lead to predictions of 1.24% [0.58, 3.14] and 2.45% [1.23, 5.9], respectively without NTEs. The major conclusion of our report is that high LET risk prediction uncertainties due to QFs parameters, NTEs, and possible increase lethality at high LET are dominant contributions to GCR uncertainties and should be the focus of space radiation research.

Evidence of rapid non-targeted effects of cycloheximide on soil bacteria using 13C-PLFA analysis

Stable isotope probing of phospholipid fatty acids (PLFA-SIP) is useful when studying bacterial contributions to soil processes, and it is an effective way to separate fungal and bacterial activity by linking 13C enrichment to specific PLFAs. Distinguishing bacterial contributions to soil processes often employs selective inhibitors; however, studies demonstrating their efficacy when using PLFA-SIP are less common. Here, we determined the effect of the fungal inhibitor cycloheximide (4.8 mg g−1 dry soil) and the bacterial inhibitor bronopol (0.48 mg g−1 dry soil) on microbial communities white spruce [Picea glauca (Moench) Voss] forest floor by measuring the uptake of 13C-enriched glucose (2 mg g−1 dry soil) in microbial PLFAs. We targeted [13C]glucose uptake by the bacterial community conditioned to a stable soil environment of 23 °C for over 2 wk rather than new bacteria generated from active colony growth caused by glucose addition. Nearly all bacterial PLFAs exhibited pronounced inhibition of 13C enrichment in the presence of bronopol. Limited inhibition of 13C enrichment in the presence of cycloheximide was observed as bacterial PLFA affected by cycloheximide had roughly one third less 13C enrichment than samples emended with [13C]glucose alone. Inhibitory effects only reduced 13C enrichment and did not affect total PLFA concentrations, implying that the inhibitors in the concentrations applied were impeding bacterial activity without causing cell death. Based on this work, we conclude that bronopol is an effective inhibitor for bacteria. Additionally, non-targeted effects of cycloheximide on soil bacteria must be accounted for when it is used in soil incubations.

Assessing Nonlinearity in Harderian Gland Tumor Induction Using Three Combined HZE-irradiated Mouse Datasets.

Recently reported studies considering nonlinearity in the effects of low-dose space radiation have assumed a nontargeted mechanism. To date, few analyses have been performed to assess whether a nontargeted term is supported by the available data. The Harderian gland data from Alpen et al. (published in 1993 and 1994), and Chang et al. (2016) provide the most diversity of ions and energies in a tumor induction model, including multiple high-energy and charge particles. These data can be used to investigate various nonlinearity assumptions against a linear model, including nontargeted effects in the low-dose region or cell sterilization at high doses. In this work, generalized linear models were used with the log complement link function to analyze the binomial data from the studies independently and combined. While there was some evidence of nonlinearity that was best described by a cell-sterilization model, the linear model was adequate to describe the data. The current data do not support the addition of a nontargeted effects term in any model. While adequate data are available in the low-dose region (<0.5 Gy) to support a nontargeted effects term if valid, additional data in the 1-2 Gy region are necessary to achieve power for cell-sterilization analysis validation. The current analysis demonstrates that the Harderian gland tumor data do not support the use of a nontargeted effects term in human cancer risk models.