Photodynamic effects of a novel pterin derivative on a pancreatic cancer cell line

6-Formylpterin (6FP) has the potential to produce singlet oxygen (1O2) under UV-A radiation. In order to apply this potential to anti-cancer photodynamic therapy (PDT), we prepared a novel variant of 6FP, 2-(N,N-dimethylaminomethyleneamino)-6-formyl-3-pivaloylpteridine-4-one (6FP-tBu-DMF), and examined its photodynamic effects on a pancreatic cancer cell line, Panc-1 cells. The study using laser scanning confocal microscopy showed that the drug uptake, the 1O2 generation, and cell death were observed in the 6FP-tBu-DMF-treated cells, while these phenomena were not observed in the 6FP-treated cells. The MTT assay also showed the decrease in cell viability only in the 6FP-tBu-DMF-treated cells. Since 6FP and 6FP-tBu-DMF generate 1O2 to the same extent under UV-A radiation in aqueous solutions, these results indicated that the differences in the photodynamic effects between 6FP and 6FP-tBu-DMF were entirely attributed to the differences in the cell permeability between them. The development of cell permeable pterin derivatives has the potential for application in PDT.     

Testing threshold and hormesis in a random effects dose-response model applied to developmental toxicity data

Here we describe a random effects threshold dose-response model for clustered binary-response data from developmental toxicity studies. For our model we assume that a hormetic effect occurs in addition to a threshold effect. Therefore, the dose-response curve is based on two components: relationships below the threshold (hormetic u-shaped model) and those above the threshold (logistic model). In the absence of hormesis and threshold effects, the estimation procedure is straightforward. We introduce score tests that are derived from a random effects hormetic-threshold dose-response model. The model and tests are applied to clustered binary data from developmental toxicity studies of animals to test for hormesis and threshold effects. We also compare the score test and likelihood ratio test to test for hormesis and threshold effects in a simulated study.     

Radiation Hormesis and Cancer

Despite the long history of radiation hormesis and the public health concerns with low-level exposures to ionizing radiation, there has been surprisingly little formal evaluation of whether hormetic effects are displayed with respect to radiation exposure and cancer incidence (i.e., reduced cancer risk at low radiation doses compared to controls, enhanced cancer risk at higher doses) until relatively recently. This paper reviews data relevant to the question of radiation hormesis and cancer with particular emphasis on experimental studies in animal models exposed to low levels of ionizing radiation. Data exist that provide evidence both consistent with and/or supportive of radiation hormesis. Other biomedical research provides potentially important mechanistic insight: low dose exposures have the capacity to activate immune function to prevent the occurrence of tumor development and metastasis; low doses of radiation have been shown to reduce mutagenic responses and induce endogenous antioxidant responses. These findings are consistent with epidemiological data suggesting an inverse relationship between background radiation and cancer incidence and with occupational epidemiological investigations in which low-dose exposure groups display markedly lower standardized mortality rates than the referent or control group.     

The Concept of Hormesis in Developmental Toxicology

Animal bioassay experiments are frequently conducted to assess the toxicity of chemicals on the developing fetus. Experiments are normally conducted at dosage levels that are much higher than human exposure levels to elicit the toxic reproductive effect of the chemical in a limited number of litters. Recently there has been much discussion on the fact that some chemicals may have beneficial effects at low doses and become toxic at high doses. This concept, known as chemical hormesis, has been the focus of attention in many investigations. Here, we consider the prevalence of hormesis in developmental toxicology and show that current design of developmental toxicity testing does not accommodate the study of hormesis. If it can be proved that some developmental toxicants may have stimulatory low dose effects, then design and analysis of developmental toxicity experiments need to be revised by the scientific community and the regulatory agencies. Using a thorough analysis of an experimental data set, we further demonstrate that in order to establish the possible hormetic effects of a chemical in reproduction, often a multiple replication of the experiment may be necessary to examine such effects. Using a trend test, we illustrate that while it is possible that one replicate of a developmental toxicity experiment with a known teratogen shows strong evidence of hormesis, other replicates may show no sign of beneficial effects at low doses.     

Bystander effects, adaptive response and genomic instability induced by prenatal irradiation

The developing human embryo and fetus undergo very radiosensitive stages during the prenatal development. It is likely that the induction of low dose related effects such as bystander effects, the adaptive response, and genomic instability would have profound effects on embryonic and fetal development. In this paper, I review what has been reported on the induction of these three phenomena in exposed embryos and fetuses. All three phenomena have been shown to occur in murine embryonic or fetal cells and structures, although the induction of an adaptive response (and also likely the induction of bystander effects) are limited in terms of when during development they can be induced and the dose or dose-rate used to treat animals in utero. In contrast, genomic instability can be induced throughout development, and the effects of radiation exposure on genome instability can be observed for long times after irradiation including through pre- and postnatal development and into the next generation of mice. There are clearly strain-specific differences in the induction of these phenomena and all three can lead to long-term detrimental effects. This is true for the adaptive response as well. While induction of an adaptive response can make fetuses more resistant to some gross developmental defects induced by a subsequent high dose challenge with ionizing radiation, the long-term effects of this low dose exposure are detrimental. The negative effects of all three phenomena reflect the complexity of fetal development, a process where even small changes in the timing of gene expression or suppression can have dramatic effects on the pattern of biological events and the subsequent development of the mammalian organism.     

Faibles doses de rayonnements : quand l’hormésis et l’effet abscopal se rencontrent…

Nuclear medicine, on its diagnostic side, provides real medical services allowed by scintigraphic examinations, clinically useful and changing patient management. However, this results in the global and heterogenous irradiation of patients, delivering prolonged and continuous exposure at low (or very low) dose rates which inevitably raises the question of possible stochastic effects… On the other hand, on its therapeutic side, nuclear medicine, using much higher activities, claims to treat or cure some cancers. This apparent contradiction can be solved by recalling some important concepts, poorly known as recent but very likely to usefully illuminate the debate on the effects of low and very low doses, and especially to introduce some consistency. The effect of ionizing radiation, at low doses, is ambivalent and must be interpreted through a dynamic equilibrium between two phenomena acting in opposite directions: on one hand, the contribution of radiations contributes to potentially deleterious lesions; on the other hand the stimulation by these same radiations of mechanisms of defense, especially immunological, against the induction of tumors. This gives them a true systemic action, which corresponds, in the case of proven cancer, by the so called “abscopal effect”, whose understanding is similar to that of the phenomenon of hormesis, in favor of a reduction in the risk of cancer at (very) low doses. These concepts undermine the RLSS (linear relationship without threshold), and open new fields of research to better understand and master the modalities of IRV (Internalized Radiotherapy Vectorized). The effects of low doses can only be considered in a dynamic and integrated way. Radiation-induced carcinogenesis is in fact a threshold effect, which cannot be extrapolated linearly below this threshold.     

Nontargeted effects of ionizing radiation: implications for low-dose exposures

The traditional thinking has been that the biological effects of ionizing radiation occur in irradiated cells as a consequence of the DNA damage they incur. This implies that: 1) biological effects occur only in irratiated cells, 2) radiation traversal through the nucleus of the cell is a prerequisite to produce a biological response, and 3) DNA is the target molecule in the cell. Evidence has been emerging, however, for non-DNA targeted effects of radiation; that is, effects including mutations, chromosomal aberrations, and changes in gene expression which occur in cells that in themselves receive no radiation exposure. Two of these phenomena will be described in this paper. The first is radiation-induced genomic instability whereby biological effects, including elevated frequencies of mutations and chromosomal aberrations, arise in the distant descendants of irradiated cells. The second phenomenon has been termed the "bystander effect", whereby in a mixed population of irradiated and nonirradiated cells, biological effects arise in those cells that receive no radiation exposure. The damage signals are transmitted from cell to cell through gap junction channels, and the genetic effects observed in bystander cells appear to result from an upregulation of oxidative stress. The possible influence of these non-targeted effects of radiation of the respounse to low-dose exposures is discussed.     

Effects of acute low doses of Gamma-radiation on erythrocytes membrane

It is believed that any dose of ionizing radiation may damage cells and that the mutated cells could develop into cancer cells. Additionally, results of research performed over the past century on the effects of low doses of ionizing radiation on biological organisms show beneficial health effects, called hormesis. Much less is known about the cellular response to low doses of ionizing radiation, such as those typical for medical diagnostic procedures, normal occupational exposures or cosmic-ray exposures at flight altitudes. Extrapolating from the effects observed at higher doses to predict changes in cells after low-dose exposure is problematic. We examined the biological effects of low doses (0.01–0.3 Gy) of γ-radiation on the membrane characteristics of erythrocytes of albino rats and carried out osmotic fragility tests and Fourier transform infrared spectroscopy (FTIR). Our results indicate that the lowest three doses in the investigated radiation range, i.e., 0.01, 0.025 and 0.05 Gy, resulted in positive effects on the erythrocyte membranes, while a dose of 0.1 Gy appeared to represent the limiting threshold dose of those positive effects. Doses higher than 0.1 Gy were associated with the denaturation of erythrocyte proteins.     

Photons – Radiobiological issues related to the risk of second malignancies

Photons are widely used in radiotherapy and while they are low LET radiation, can still pose a risk in developing second malignant neoplasms (SMN). Due to the physics of photons that allow distribution of energy outside the target volume, out-of-field irradiation is an important component of SMN risk assessment. The epidemiological evidence supporting this risk should be augmented with radiobiological justifications for a better understanding of the underlying processes.There are several factors that impact second cancer risk which can be analysed from a radiobiological perspective: age at irradiation, type of irradiated tissue, irradiated volume, treatment technique, previous irradiation/radiological investigations. Age-dependence has a radiobiological foundation given by the higher radiosensitivity of children as compared to adult patients. However, in its 2013 report, UNSCEAR advises against generalisation of the effects of childhood radiation exposure, given the fact that these effects are strongly organ dependent. Furthermore, the age-dependent radiation sensitivity has a bimodal distribution, since aging cells present an increase in the oxidative stress, which can promote premalignant cells.Non-targeted effects such as radiation-induced genomic instability, bystander or abscopal effects could also impact on the risk of SMN. Recent studies show that beside the known cellular changes, bystander effects can be manifested through increased cell proliferation, which could be a culprit for SMN development. Furthermore, new evidence on the existence of tumour-specific cancer stem cells that are long-lived and more quiescent and radioresistant than non-stem cancer cells can raise questions about their association with SMN risk.     

Early Life Hormetic Treatments Decrease Irradiation-Induced Oxidative Damage,Increase Longevity,and Enhance Sexual Performance during Old Age in the Caribbean Fruit Fly

Early life events can have dramatic consequences on performance later in life. Exposure to stressors at a young age affects development, the rate of aging, risk of disease, and overall lifespan. In spite of this, mild stress exposure early in life can have beneficial effects on performance later in life. These positive effects of mild stress are referred to as physiological conditioning hormesis. In our current study we used anoxia conditioning hormesis as a pretreatment to reduce oxidative stress and improve organismal performance, lifespan, and healthspan of Caribbean fruit flies. We used gamma irradiation to induce mild oxidative damage in a low-dose experiment, and massive oxidative damage in a separate high-dose experiment, in pharate adult fruit flies just prior to adult emergence. Irradiation-induced oxidative stress leads to reduced adult emergence, flight ability, mating performance, and lifespan. We used a hormetic approach, one hour of exposure to anoxia plus irradiation in anoxia, to lower post-irradiation oxidative damage. We have previously shown that this anoxic-conditioning treatment elevates total antioxidant capacity and lowers post-irradiation oxidative damage to lipids and proteins. In this study, conditioned flies had lower mortality rates and longer lifespan compared to those irradiated without hormetic conditioning. As a metric of healthspan, we tracked mating both at a young age (10 d) and old age (30 d). We found that anoxia-conditioned male flies were more competitive at young ages when compared to unconditioned irradiation stressed male flies, and that the positive effects of anoxic conditioning hormesis on mating success were even more pronounced in older males. Our data shows that physiological conditioning hormesis at a young age, not only improves immediate metrics of organismal performance (emergence, flight, mating), but the beneficial effects also carry into old age by reducing late life oxidative damage and improving lifespan and healthspan.     

Pulsed microwave induced light, sound, and electrical discharge enhanced by a biopolymer

Intense flashes of light were observed in sodium bicarbonate and hydrogen peroxide solutions when they were exposed to pulsed microwave radiation, and the response was greatly enhanced by a microwave-absorbing, biosynthesized polymer, diazoluminomelanin. A FPS-7B radar transmitter, operating at 1.25 GHz provided pulses of 5.73 +/- 0.09 micros in duration at 10.00 +/- 0.03 pulses/s with 2.07 +/- 0.08 MW forward power (mean +/- standard deviation), induced the effect but only when the appropriate chemical interaction was present. This phenomenon involves acoustic wave generation, bubble formation, pulsed luminescence, ionized gas ejection, and electrical discharge. The use of pulsed microwave radiation to generate highly focused energy deposition opens up the possibility of a variety of biomedical applications, including targeting killing of microbes or eukaryotic cells. The full range of microwave intensities and frequencies that induce these effects has yet to be explored and, therefore, the health and safety implications of generating the phenomena in living tissues remain an open question.     

Effects of 50 Hz magnetic field on cell cycle kinetics and the colony forming ability of budding yeast exposed to ultraviolet radiation.

To investigate the effects of extremely low frequency magnetic fields on ultraviolet radiation (UV) exposed budding yeast, haploid yeast (Saccharomyces cerevisiae) cells of the strain SEy2101a were exposed to 50 Hz sine wave magnetic field (MF) of 120 microT with simultaneous exposure to UV radiation. Most of the UV energy was in the UVB range (280-320 nm). The biologically weighted (CIE action spectrum) dose level for the UV radiation was 175 J/m2. We examined whether 50 Hz MF affected the ability of UV irradiated yeast cells to form colonies (Colony Forming Units, CFUs). In addition, the effect of coexposure on cell cycle kinetics was investigated. Although the significant effect of MF on the cell cycle phases of UV exposed yeast cells was seen only at one time point, the overall results showed that MF exposure may influence the cell cycle kinetics at the first cycle after UV irradiation. The effect of our particular MF exposure on the colony forming ability of the UV irradiated yeast cells was statistically significant 420 min after UV irradiation. Moreover, at 240, 360, and 420 min after UV irradiation, there were fewer CFUs in every experiment in (UV+MF) exposed populations than in only UV exposed yeast populations. These results could indicate that MF exposure in conjunction with UV may have some effects on yeast cell survival or growth.     

Refinement of the dichlorofluorescein assay for flow cytometric measurement of reactive oxygen species in irradiated and bystander cell populations

Reactive oxygen species (ROS) have been implicated in many ionizing radiation-related phenomena, including bystander effects. The oxidation of 2'7'-dichlorofluorescin (DCFH) to fluorescent 2'7'-dichlorofluorescein (DCF) is commonly used for the detection of radiation-induced ROS. The DCF assay was adapted for efficient, systematic flow cytometry quantification of low-linear energy transfer (LET) gamma-radiation-induced ROS in vitro in Chinese hamster ovary (CHO) cells. This method is optimized for increased sensitivity to radiation-induced ROS and to discriminate against measurement of extracellular ROS. This method can detect a significant increase in ROS in cells exposed to gamma radiation at doses as low as 10 cGy. The antioxidants N-acetyl-cysteine and ascorbic acid (vitamin C) significantly reduced the amount of ROS measured in cells exposed to 5 Gy ionizing radiation. This method was used to measure the intracellular ROS in unirradiated CHO bystander cells co-cultured with low-LET-irradiated cells. No increase in ROS was measured in bystander cell populations co-cultured with the irradiated cells beginning 9 s after radiation exposure.     

Effect of low-dose irradiation on the lifespan in various strains of Drosophila melanogaster

Drosophila melanogaster strains defective in repair, antioxidant defense, and apoptosis displayed a higher aging rate than the wild-type strains when unexposed to ionizing radiation. Irradiation changed the lifespan depending on the genotype. The lifespan and the functional (neuromuscular) activity, which reflects the "life quality", changed in the same direction. A mechanism was suggested for the remote effect of low-dose irradiation on the lifespan. Since cells with a weakened defense system accumulate lesions and age at a higher rate, their elimination in early ontogeny decelerates age-related changes and decreases the aging rate. In subsequent generations, this somatic stress response (hormesis) is replaced by negative genetic effects at the population level and, consequently, the lifespan decreases.     

THE HORMESIS OF THE GREEN MACROALGA ULVA FASCIATA WITH LOW‐DOSE 60COBALT GAMMA RADIATION1

Homogenous germlings of the marine macroalga Ulva fasciata D. (synonym, Ulva lactuca L.) were used to study hormesis effects in macroalgae grown under a low dose of ⁶⁰Co γ‐ray radiation. The results of this study are the first to confirm the effects of macroalgal hormesis. Here it was demonstrated that growth of U. fasciata germlings was promoted substantially under 15 Gy of ⁶⁰Co γ‐ray radiation, with an average increase of algal biomass of 47.43%. The levels of polysaccharides and lipids varied among the tested material and showed no effects from the ⁶⁰Co γ‐ray radiation. However, the amount of protein was higher in the irradiated algae than in the control; the highest protein content of the irradiated algae was 3.958% (dry weight), in contrast to 2.318% in nonirradiated samples. This technique was applied to a field algal mass culture, which decreased the harvest time from 90 to 60 d. The mass culture approach may facilitate the production of macroalgae under unstable weather conditions such as typhoons in the summer or strong waves in the winter. The mass‐cultured macroalgae could be used as a source of bioenergy through the fermentation of algal simple sugars that derived from polysaccharides to produce ethanol.     

Slowing down aging from within: mechanistic aspects of anti-aging hormetic effects of mild heat stress on human cells

Since aging is primarily the result of a failure of maintenance and repair mechanisms, various approaches are being developed in order to stimulate these pathways and modulate the process of aging. One such approach, termed hormesis, involves challenging cells and organisms by mild stress that often results in anti-aging and life prolonging effects. In a series of experimental studies, we have reported that repeated mild heat stress (RMHS) has anti-aging hormetic effects on growth and various cellular and biochemical characteristics of human skin fibroblasts undergoing aging in vitro. These beneficial effects of repeated challenge include the maintenance of stress protein profile, reduction in the accumulation of oxidatively and glycoxidatively damaged proteins, stimulation of the proteasomal activities for the degradation of abnormal proteins, improved cellular resistance to other stresses, and enhanced levels of cellular antioxidant ability. In order to elucidate the molecular mechanisms of hormetic effects of RMHS, we are now undertaking studies on signal transduction pathways, energy production and utilisation kinetics, and the proteomic analysis of patterns of proteins synthesised and their posttranslational modifications in various types of human cells undergoing cellular aging in vitro. Human applications of hormesis include early intervention and modulation of the aging process to prevent or delay the onset of age-related conditions, such as sarcopenia, Alzheimer's disease, Parkinson's disease, cataracts and osteoporosis.     

The link between physics and chemistry in track modelling

The physical structure of a radiation track provides the initial conditions for the modelling of radiation chemistry. These initial conditions are not perfectly understood, because there are important gaps between what is provided by a typical track structure model and what is required to start the chemical model. This paper addresses the links between the physics and chemistry of tracks, with the intention of identifying those problems that need to be solved in order to obtain an accurate picture of the initial conditions for the purposes of modelling chemistry. These problems include the reasons for the increased yield of ionisation relative to homolytic bond breaking in comparison with the gas phase. A second area of great importance is the physical behaviour of low-energy electrons in condensed matter (including thermolisation and solvation). Many of these processes are not well understood, but they can have profound effects on the transient chemistry in the track. Several phenomena are discussed, including the short distance between adjacent energy loss events, the molecular nature of the underlying medium, dissociative attachment resonances and the ability of low-energy electrons to excite optically forbidden molecular states. Each of these phenomena has the potential to modify the transient chemistry substantially and must therefore be properly characterised before the physical model of the track can be considered to be complete. Received: 15 March 1999 / Accepted in revised form: 29 July 1999     

Altruistic cell suicide in relation to radiation hormesis

The high radiosensitivity to killing of undifferentiated primordial cells (Bergonié and Tribondeau 1906) can be described as a manifestation of the suicide of injured cells for the benefit of an organism as a whole if their suicide stimulates proliferation of healthy cells to replace them, resulting in complete elimination of injury. This process is called cell-replacement repair, to distinguish it from DNA repair which is rarely complete. 'Cell suicide', 'programmed death' and 'apoptosis' are terms used for the same type of active cell death. Cell suicide is not always altruistic. Altruistic suicide in Drosophila, mice, humans, plants, and E. coli is reviewed in this paper to illustrate its widely different facets. The hypothesis that in animals, radiation hormesis results from altruistic cell suicide is proposed. This hypothesis can explain the hormetic effect of low doses of radiation on the immune system in mice. In contrast, in plants, radiation hormesis seems to be mainly due to non-altruistic cell death. HORMESIS--'the stimulating effect of small doses of substances which in larger doses are inhibitory' (British Medical Dictionary, Caxton Publ. Co., 1961).     

Effect of low-dose irradiation on the lifespan in various strains of Drosophila melanogaster

Drosophila melanogaster strains defective in repair, antioxidant defense, and apoptosis displayed a higher aging rate than the wild-type strains when unexposed to ionizing radiation. Irradiation changed the lifespan depending on the genotype. The lifespan and the functional (neuromuscular) activity, which reflects the “life quality,” changed in the same direction. A mechanism was suggested for the remote effect of low-dose irradiation on the lifespan. Since cells with a weakened defense system accumulate lesions and age at a higher rate, their elimination in early ontogeny decelerates age-related changes and decreases the aging rate. In subsequent generations, this somatic stress response (hormesis) is replaced by negative genetic effects at the population level and, consequently, the lifespan decreases.     

Characteristics of DNA-binding proteins determine the biological sensitivity to high-linear energy transfer radiation

Non-homologous end-joining (NHEJ) and homologous recombination repair (HRR), contribute to repair ionizing radiation (IR)-induced DNA double-strand breaks (DSBs). Mre11 binding to DNA is the first step for activating HRR and Ku binding to DNA is the first step for initiating NHEJ. High-linear energy transfer (LET) IR (such as high energy charged particles) killing more cells at the same dose as compared with low-LET IR (such as X or γ rays) is due to inefficient NHEJ. However, these phenomena have not been demonstrated at the animal level and the mechanism by which high-LET IR does not affect the efficiency of HRR remains unclear. In this study, we showed that although wild-type and HRR-deficient mice or DT40 cells are more sensitive to high-LET IR than to low-LET IR, NHEJ deficient mice or DT40 cells are equally sensitive to high- and low-LET IR. We also showed that Mre11 and Ku respond differently to shorter DNA fragments in vitro and to the DNA from high-LET irradiated cells in vivo. These findings provide strong evidence that the different DNA DSB binding properties of Mre11 and Ku determine the different efficiencies of HRR and NHEJ to repair high-LET radiation induced DSBs.