Self-assembly of polyisoprenoyl gemcitabine conjugates: influence of supramolecular organization on their biological activity.

An amphiphilic prodrug of gemcitabine, a cytidine analogue used clinically against various tumors, had been previously synthesized by covalent coupling to squalene, a natural isoprenoid chain. The resulting bioconjugate self-assembled spontaneously in water as nanoparticles, displaying an impressive activity both in vitro and in vivo. The aim of the present study was to determine the influence of the length of the isoprene moiety on the structure of the nanoparticles, in an attempt to establish a relationship between the chemical structure of the prodrug, its supramolecular organization, and its pharmacological activity. Remarkably, gemcitabine-squalene and gemcitabine-5-isoprenes, which differ only in the position of two methyl groups on the hydrophobic chain, displayed different supramolecular organizations and different anticancer activities on some cell lines. This difference in activity was related to the ability of nanoparticles to be internalized by cells.

Activity of immunoproteasome inhibitor ONX-0914 in acute lymphoblastic leukemia expressing MLL-AF4 fusion protein.

Proteasome inhibitors bortezomib and carfilzomib are approved for the treatment of multiple myeloma and mantle cell lymphoma and have demonstrated clinical efficacy for the treatment of acute lymphoblastic leukemia (ALL). The t(4;11)(q21;q23) chromosomal translocation that leads to the expression of MLL-AF4 fusion protein and confers a poor prognosis, is the major cause of infant ALL. This translocation sensitizes tumor cells to proteasome inhibitors, but toxicities of bortezomib and carfilzomib may limit their use in pediatric patients. Many of these toxicities are caused by on-target inhibition of proteasomes in non-lymphoid tissues (e.g., heart muscle, gut, testicles). We found that MLL-AF4 cells express high levels of lymphoid tissue-specific immunoproteasomes and are sensitive to pharmacologically relevant concentrations of specific immunoproteasome inhibitor ONX-0914, even in the presence of stromal cells. Inhibition of multiple active sites of the immunoproteasomes was required to achieve cytotoxicity against ALL. ONX-0914, an inhibitor of LMP7 (ß5i) and LMP2 (ß1i) sites of the immunoproteasome, and LU-102, inhibitor of proteasome ß2 sites, exhibited synergistic cytotoxicity. Treatment with ONX-0914 significantly delayed the growth of orthotopic ALL xenograft tumors in mice. T-cell ALL lines were also sensitive to pharmacologically relevant concentrations of ONX-0914. This study provides a strong rationale for testing clinical stage immunoproteasome inhibitors KZ-616 and M3258 in ALL.

Strontium-90 and caesium-137 activity concentrations in bats in the Chernobyl exclusion zone.

Bats are a protected species and as such may be an object of protection in radiological assessments of the environment. However, there have previously been only few radioecological studies of species of bats. In this paper, results for >140 measurements of (90)Sr and (137)Cs in 10 species of bats collected within the Chernobyl zone are presented. There was some indication of a decreasing transfer of (90)Sr with increasing deposition, although this was inconsistent across species and explained little of the observed variability. There was no difference between male and female bats in the transfer (expressed as the ratio of whole-body activity concentrations to those in soil) of either radionuclide. There was considerable variability in transfer across all species groups. At two sites where there were sufficient data, Eptesicus serotinus was found to have higher transfer than other species.

X-ray microfluorescence for biodistribution studies of nanomedicines.

Currently, the in vivo distribution of drugs is investigated by non-spatial quantitative techniques. With the emergence of personal therapies using nanomedicines, deeper investigations are required to precisely know the in vivo fate of entrapped drugs, especially to predict possible toxicity. Here, we assess the capabilities of SR-µXRF for i) detecting drugs into nanomedicines without adding any marker, ii) mapping their distribution into tissues and iii) locally quantifying the drugs loaded into nanomedicines. To prepare the nanomedicine model, we used the bioconjugate diamine(dichloro)platinum (SQ-CDD) developed in the TERNANOMED Grant Project. Nanomedicines were intravenously injected into a nude mice model bearing a pancreatic tumour (PANC-1). The X-ray microfluorescence experiments were performed on embeds tissue sections of kidney and tumor at 2h and 24h after nanoparticles injection. Data collection was performed on the micro-imaging beamline ID13 of the European Synchrotron Radiation Facility (ESRF). A quantitative study was performed by atomic absorption spectroscopy (AAS), allowing to compare the platinum concentrations with those measured by X-ray. This study shows that the synchrotron radiation-based µXRF analysis is sensitive enough to detect and map the distribution of a drug entrapped into nanomedicine. A quantitative local analysis is possible with a tissue element as reference, or semi-quantitatively if the tissue reference is not homogenous.

Positively charged cyclodextrins as effective molecular transporters of active phosphorylated forms of gemcitabine into cancer cells.

Positively charged cyclodextrins (PCCDs) are molecular carriers of particular interest for their ability to readily enter into cancer cells. Of main interest, guanidino- and aminoalkyl- PCCDs can be conveniently synthesized and form stable and strong inclusion complexes with various active molecules bearing phosphate groups. We have addressed here the challenge to deliver into cancer cells phosphorylated gemcitabine drugs well known for their instability and inability to permeate cell membranes. NMR data corroborated by semiempirical theoretical calculations have shown that aminoalkyl-CDs form sufficiently stable complexes with both mono- and tri-phosphate forms of gemcitabine by simple mixing of the compounds in aqueous solution at physiological pH. Confocal microscopy and radioactivity counting experiments revealed that the developed systems enabled phosphorylated gemcitabine to penetrate efficiently into aggressive human breast cancer cells (MCF7), eventually leading to a substantial reduction of IC50 values. Moreover, compared to free drugs, phosphorylated metabolites of gemcitabine encapsulated in PCCDs displayed improved in vitro activities also on the aggressive human cancer cells CCRF-CEM Ara-C/8 C, a nucleoside transport-deficient T leukemia cell line. The current study offers the proof-of-principle that phosphorylated nucleoside drugs could be efficiently transported by PCCDs into cancer cells.

The transfer of (137)Cs, Pu isotopes and (90)Sr to bird, bat and ground-dwelling small mammal species within the Chernobyl exclusion zone.

Protected species are the focus of many radiological environmental assessments. However, the lack of radioecological data for many protected species presents a significant international challenge. Furthermore, there are legislative restrictions on destructive sampling of protected species to obtain such data. Where data are not available, extrapolations are often made from 'similar' species but there has been little attempt to validate this approach. In this paper we present what, to our knowledge, is the first study purposefully designed to test the hypothesis that radioecological data for unprotected species can be used to estimate conservative radioecolgical parameters for protected species; conservatism being necessary to ensure that there is no significant impact. The study was conducted in the Chernobyl Exclusion Zone. Consequently, we are able to present data for Pu isotopes in terrestrial wildlife. There has been limited research on Pu transfer to terrestrial wildlife which contrasts with the need to assess radiation exposure of wildlife to Pu isotopes around many nuclear facilities internationally. Our results provide overall support for the hypothesis that data for unprotected species can be used to adequately assess the impacts for ionising radiation on protected species. This is demonstrated for a range of mammalian and avian species. However, we identify one case, the shrew, for which data from other ground-dwelling small mammals would not lead to an appropriately conservative assessment of radiation impact. This indicates the need to further test our hypothesis across a range of species and ecosystems, and/or ensure adequate conservatism within assessments. The data presented are of value to those trying to more accurately estimate the radiation dose to wildlife in the Chernobyl Exclusion Zone, helping to reduce the considerable uncertainty in studies reporting dose-effect relationships for wildlife. A video abstract for this paper is available from: http://bit.ly/1JesKPc.

Effects of radionuclide contamination on leaf litter decomposition in the Chernobyl exclusion zone.

The effects of radioactive contamination on ecosystem processes such as litter decomposition remain largely unknown. Because radionuclides accumulated in soil and plant biomass can be harmful for organisms, the functioning of ecosystems may be altered by radioactive contamination. Here, we tested the hypothesis that decomposition is impaired by increasing levels of radioactivity in the environment by exposing uncontaminated leaf litter from silver birch and black alder at (i) eleven distant forest sites differing in ambient radiation levels (0.22-15µGyh(-1)) and (ii) along a short distance gradient of radioactive contamination (1.2-29µGyh(-1)) within a single forest in the Chernobyl exclusion zone. In addition to measuring ambient external dose rates, we estimated the average total dose rates (ATDRs) absorbed by decomposers for an accurate estimate of dose-induced ecological consequences of radioactive pollution. Taking into account potential confounding factors (soil pH, moisture, texture, and organic carbon content), the results from the eleven distant forest sites, and from the single forest, showed increased litter mass loss with increasing ATDRs from 0.3 to 150µGyh(-1). This unexpected result may be due to (i) overcompensation of decomposer organisms exposed to radionuclides leading to a higher decomposer abundance (hormetic effect), and/or (ii) from preferred feeding by decomposers on the uncontaminated leaf litter used for our experiment compared to locally produced, contaminated leaf litter. Our data indicate that radio-contamination of forest ecosystems over more than two decades does not necessarily have detrimental effects on organic matter decay. However, further studies are needed to unravel the underlying mechanisms of the results reported here, in order to draw firmer conclusions on how radio-contamination affects decomposition and associated ecosystem processes.

Overview of the cooperation between the Chernobyl Center's International Radioecology Laboratory in Slavutych, Ukraine, and U.S. research centers between 2000 and 2010.

The International Radioecology Laboratory (IRL) located in Slavutych, Ukraine, was created in 1999 under the initiative of the United States Government and the Government of Ukraine in the framework of international cooperation on evaluation and minimization of consequences of the Chernobyl nuclear power plant (ChNPP) accident. Since the time the IRL was founded, it has participated in a large number of projects, including the following: 1) study of radionuclide accumulation, distribution, and migration in components of various ecological systems of the Chernobyl Exclusion Zone (ChEZ); 2) radiation dose assessments; 3) study of the effects of radiation influence on biological systems; 4) expert analysis of isotopic and quantitative composition of radioactive contaminants; 5) development of new methods and technologies intended for radioecological research; 6) evaluation of future developments and pathways for potential remediation of the ChEZ areas; 7) assistance in provision of physical protection systems for ionizing irradiation sources at Ukrainian enterprises; 8) reviews of open Russian language publications on issues associated with consequences of the ChNPP accident, radioactive waste management, radioecological monitoring, and ChNPP decommissioning; 9) conduct of training courses on problems of radioecology, radiation safety, radioecological characterization of test sites and environmental media, and research methods; 10) conduct of on-site scientific conferences and workshops on the ChEZ and radioecology problems; participation in off-site scientific conferences and meetings; and 11) preparation of scientific and popular science publications and interactions with mass media representatives. This article provides a brief overview of the major achievements resulting from this cooperation between the IRL and U.S. research centers.

Method for simultaneous 90Sr and 137Cs in-vivo measurements of small animals and other environmental media developed for the conditions of the Chernobyl exclusion zone.

To perform in vivo simultaneous measurements of the 90Sr and 137Cs content in the bodies of animals living in the Chernobyl Exclusion Zone (ChEZ), an appropriate method and equipment were developed and installed in a mobile gamma beta spectrometry laboratory. This technique was designed for animals of relatively small sizes (up to 50 g). The 90Sr content is measured by a beta spectrometer with a 0.1-mm-thick scintillation plastic detector. The spectrum processing takes into account the fact that the measured object is "thick-layered" and contains a comparable quantity of 137Cs, which is a characteristic condition of the ChEZ. The 137Cs content is measured by a NaI scintillation detector that is part of the combined gamma beta spectrometry system. For environmental research performed in the ChEZ, the advantages of this method and equipment (rapid measurements, capability to measure live animals directly in their habitat, and the capability of simultaneous 90Sr and 137Cs measurements) far outweigh the existing limitations (considerations must be made for background radiation and the animal size, skeletal shape, and body mass). The accuracy of these in vivo measurements is shown to be consistent with standard spectrometric and radiochemical methods. Apart from the in vivo measurements, the proposed methodology, after a very simple upgrade that is also described in this paper, works even more accurately with samples of other media, such as soil and plants.

Radiation ecology issues associated with murine rodents and shrews in the Chernobyl exclusion zone.

This article describes major studies performed by the Chernobyl Center's International Radioecology Laboratory (Slavutich, Ukraine) on radioecology of murine rodents and shrews inhabiting the Chernobyl Exclusion Zone. The article addresses the long-term (1986-2005) and seasonal dynamics of radioactive contamination of animals and reviews interspecies differences in radionuclide accumulations and factors affecting the radionuclide accumulations. It is shown that bioavailability of radionuclides in the "soil-to-plant" chain and a trophic specialization of animals play key roles in determining their actual contamination levels. The total absorbed dose rates in small mammals significantly reduced during the years following the Chernobyl Nuclear Power Plant accident. In 1986, the absorbed dose rate reached 1.3-6.0 Gy h(-1) in the central areas of the Chernobyl Exclusion Zone (the "Red Forest"). In 1988 and 1990, the total absorbed dose rates were 1.3 and 0.42 Gy h(-1), respectively. In 1995, 2000, and 2005, according to the present study, the total absorbed dose rates rarely exceeded 0.00023, 0.00018, and 0.00015 Gy h(-1), respectively. Contributions of individual radiation sources into the total absorbed dose are described.

Radiation dose assessment for the biota of terrestrial ecosystems in the shoreline zone of the Chernobyl nuclear power plant cooling pond.

Radiation exposure of the biota in the shoreline area of the Chernobyl Nuclear Power Plant Cooling Pond was assessed to evaluate radiological consequences from the decommissioning of the Cooling Pond. This paper addresses studies of radioactive contamination of the terrestrial faunal complex and radionuclide concentration ratios in bodies of small birds, small mammals, amphibians, and reptiles living in the area. The data were used to calculate doses to biota using the ERICA Tool software. Doses from 90Sr and 137Cs were calculated using the default parameters of the ERICA Tool and were shown to be consistent with biota doses calculated from the field data. However, the ERICA dose calculations for plutonium isotopes were much higher (2-5 times for small mammals and 10-14 times for birds) than the doses calculated using the experimental data. Currently, the total doses for the terrestrial biota do not exceed maximum recommended levels. However, if the Cooling Pond is allowed to draw down naturally and the contaminants of the bottom sediments are exposed and enter the biological cycle, the calculated doses to biota may exceed the maximum recommended values. The study is important in establishing the current exposure conditions such that a baseline exists from which changes can be documented following the lowering of the reservoir water. Additionally, the study provided useful radioecological data on biota concentration ratios for some species that are poorly represented in the literature.

Vertical migration of radionuclides in the vicinity of the chernobyl confinement shelter.

Studies of vertical migration of Chernobyl-origin radionuclides in the 5-km zone of the Chernobyl Nuclear Power Plant (ChNPP) in the area of the Red Forest experimental site were completed. Measurements were made by gamma spectrometric methods using high purity germanium (HPGe) detectors with beryllium windows. Alpha-emitting isotopes of plutonium were determined by the measurement of the x-rays from their uranium progeny. The presence of 60Co, 134,137Cs, 154,155Eu, and 241Am in all soil layers down to a depth of 30 cm was observed. The presence of 137Cs and 241Am was noted in the area containing automorphous soils to a depth of 60 cm. In addition, the upper soil layers at the test site were found to contain 243Am and 243Ñm. Over the past 10 years, the 241Am/137Cs ratio in soil at the experimental site has increased by a factor of 3.4, nearly twice as much as would be predicted based solely on radioactive decay. This may be due to "fresh" fallout emanating from the ChNPP Confinement Shelter.

Assessment of the radionuclide composition of "hot particles" sampled in the Chernobyl nuclear power plant fourth reactor unit.

Fuel-containing materials sampled from within the Chernobyl Nuclear Power Plant (ChNPP) Unit 4 Confinement Shelter were spectroscopically studied for gamma and alpha content. Isotopic ratios for cesium, europium, plutonium, americium, and curium were identified, and the fuel burn-up in these samples was determined. A systematic deviation in the burn-up values based on the cesium isotopes in comparison with other radionuclides was observed. The studies conducted were the first ever performed to demonstrate the presence of significant quantities of 242Cm and 243Cm. It was determined that there was a systematic underestimation of activities of transuranic radionuclides in fuel samples from inside of the ChNPP Confinement Shelter, starting from 241Am (and going higher) in comparison with the theoretical calculations.

Frequency distributions of 90Sr and 137Cs concentrations in an ecosystem of the "Red Forest" area in the Chernobyl exclusion zone.

In the most highly contaminated region of the Chernobyl Exclusion Zone, the "Red Forest" site, the accumulation of the major dose-affecting radionuclides (90Sr and 137Cs) within the components of an ecological system encompassing 3,000 m(2) was characterized. The sampled components included soils (top 0-10 cm depth), Molina caerulea (blue moor grass), Camponotus vagus (carpenter ants), and Pelobates fuscus (spade-footed toad). In a comparison among the components of this ecosystem, the 90Sr and 137Cs concentrations measured in 40 separate grids exhibited significant differences, while the frequency distribution of the values was close to a logarithmically-normal leptokurtic distribution with a significant right-side skew. While it is important to identify localized areas of high contamination or "hot spots," including these values in the arithmetic mean may overestimate the exposure risk. In component sample sets that exhibited logarithmically normal distribution, the geometric mean more accurately characterizes a site. Ideally, risk assessment is most confidently achieved when the arithmetic and geometric means are most similar, meaning the distribution approaches normal. Through bioaccumulation, the highest concentrations of 90Sr and 137Cs were measured in the blue moor grass and spade-footed toad. These components also possessed distribution parameters that shifted toward a normal distribution.

Environmental radiation monitoring in the Chernobyl exclusion zone--history and results 25 years after.

This paper describes results of the radiation environmental monitoring performed in the Chernobyl Exclusion Zone (ChEZ) during the period following the 1986 Chernobyl Nuclear Power Plant accident. This article presents a brief overview of five comprehensive reports generated under Contract No. DE-AC09-96SR18500 (Washington Savannah River Company LLC, Subcontract No. AC55559N, SOW No. ON8778) and summarizes characteristics of the ChEZ and its post-accident status. The history of development of the radiation monitoring research in the ChEZ is described also. This paper addresses the characteristics of radiation monitoring in the ChEZ, its major goals and objectives, and changes in these goals and objectives in the course of time, depending on the tasks associated with the phase of mitigation of the ChNPP accident consequences. The results of the radiation monitoring in the ChEZ during the last 25 years are also provided.