Dosimetry of the PIM1 Pion Beam at the Paul Scherrer Institute for Radiobiological Studies of Mice.

Significant past work has identified unexpected risks of central nervous system (CNS) exposure to the space radiation environment, where long-lasting functional decrements have been associated with multiple ion species delivered at low doses and dose rates. As shielding is the only established intervention capable of limiting exposure to the dangerous radiation fields in space, the recent discovery that pions, emanating from regions of enhanced shielding, can contribute significantly to the total absorbed dose on a deep space mission poses additional concerns. As a prerequisite to biological studies evaluating pion dose equivalents for various CNS exposure scenarios of mice, a careful dosimetric validation study is required. Within our ultimate goal of evaluating the functional consequences of defined pion exposures to CNS functionality, we report in this article the detailed dosimetry of the PiMI pion beam line at the Paul Scherrer Institute, which was developed in support of radiobiological experiments. Beam profiles and contamination of the beam by protons, electrons, positrons and muons were characterized prior to the mice irradiations. The dose to the back and top of the mice was measured using thermoluminescent dosimeters (TLD) and optically simulated luminescence (OSL) to cross-validate the dosimetry results. Geant4 Monte Carlo simulations of radiation exposure of a mouse phantom in water by charged pions were also performed to quantify the difference between the absorbed dose from the OSL and TLD and the absorbed dose to water, using a simple model of the mouse brain. The absorbed dose measured by the OSL dosimeters and TLDs agreed within 5-10%. A 30% difference between the measured absorbed dose and the dose calculated by Geant4 in the dosimeters was obtained, probably due to the approximated Monte Carlo configuration compared to the experiment. A difference of 15-20% between the calculated absorbed dose to water at a 5 mm depth and in the passive dosimeters was obtained, suggesting the need for a correction factor of the measured dose to obtain the absorbed dose in the mouse brain. Finally, based on the comparison of the experimental data and the Monte Carlo calculations, we consider the dose measurement to be accurate to within 15-20%.

COMPARISON OF PADC NEUTRON DETECTORS FROM DIFFERENT SUPPLIERS.

The objective of this study was to compare poly-allyl diglycol carbonate (PADC) track detectors from different suppliers for linear energy transfer (LET) spectrometry and neutron dosimetry. PADCs are commonly used for passive personal neutron dosimetry, where a common approach is to couple the PADC with a plastic radiator to generate secondary charged particles. The neutron dose can be estimated using the track density or the average LET of the secondary particles. The characterisation of PADC in terms of LET spectrometric capability and neutron sensitivity was previously performed using PADC from Intercast S.r.l., Parma, Italy, but this company stopped the production. Since it is common experience that material from different suppliers can have different properties, it became necessary to perform a comparison of PADCs from different suppliers with the Intercast material. The study permits to compare the reading procedures used at Politecnico di Milano and at the Paul Scherrer Institute.

COMPARISON BETWEEN PADC AND FNTD NEUTRON DETECTOR SYSTEMS IN BLIND TESTS.

The objective of this study was to compare a neutron dosimetry system based on polyallyl diglycol carbonate (PADC) detectors with a new system based on Al2O3:C,Mg fluorescence nuclear track detectors (FNTD). The irradiations, performed as part of an intercomparison organized by the Physikalisch-Technische Bundesanstalt (PTB), Germany, were on a PMMA phantom with 252Cf or 241Am-Be source, usually with the phantom surface perpendicular to the radiation beam (0° angle), and with Hp(10) values between 0.3 and 7 mSv. One 252Cf irradiation was performed at 30° angle, and one with an additional 1 mSv gamma irradiation. The results showed an agreement between the two techniques with an average and maximum difference between PADCs and FNTDs of 1.5 and 22%, respectively, if one compares only cases of doses >1 mSv. For one of the irradiation conditions with dose of 0.9 mSv, use of the incorrect calibration factor for the FNTD (252Cf instead of 241Am-Be) led to reported values ~×2 larger than the given doses, due to low statistics in the determination of the ratio between 6Li-doped glass and polyethylene neutron converters. Although the FNTD track analysis algorithm may need further development, the results presented here demonstrate the feasibility of the FNTD technology and indicate areas requiring improvements.

HIGH DOSE FAST NEUTRON DOSIMETRY USING PADC PLASTIC NUCLEAR TRACK DETECTORS AND GREY LEVEL ANALYSIS.

The objective of this work is to demonstrate the possibility of performing fast neutron dosimetry up to 5 Sv using optical absorbance of polyallyl diglycol carbonate (PADC) detectors, obtained through grey level analysis of PADC images acquired with a commercial track-counting dosimetry system, and estimate the uncertainties involved. PADCs were irradiated with doses from 100 mSv to 5 Sv (252Cf source) and etched. PADC images were acquired using the TASLIMAGE™ Neutron Dosimetry System (Track Analysis Systems Ltd.) and analysed to obtain the grey levels and the optical absorbance. The absorbance from different detectors and batches was analysed to determine the uncertainties involved, from which the final uncertainty in the method, ~30% and dominated by the uncertainty in the calibration curve, was estimated. A dose estimation <2 Sv can also be performed using a 'universal curve' by normalising the absorbance to that of a detector irradiated with 1 Sv. The data presented here allows the extension of the dose range of track counting systems using no additional equipment, only the images already acquired by the systems.

Transplant tolerance to pancreatic islets is initiated in the graft and sustained in the spleen.

The immune system is comprised of several CD4(+) T regulatory (Treg) cell types, of which two, the Foxp3(+) Treg and T regulatory type 1 (Tr1) cells, have frequently been associated with transplant tolerance. However, whether and how these two Treg-cell types synergize to promote allograft tolerance remains unknown. We previously developed a mouse model of allogeneic transplantation in which a specific immunomodulatory treatment leads to transplant tolerance through both Foxp3(+) Treg and Tr1 cells. Here, we show that Foxp3(+) Treg cells exert their regulatory function within the allograft and initiate engraftment locally and in a non-antigen (Ag) specific manner. Whereas CD4(+) CD25(-) T cells, which contain Tr1 cells, act from the spleen and are key to the maintenance of long-term tolerance. Importantly, the role of Foxp3(+) Treg and Tr1 cells is not redundant once they are simultaneously expanded/induced in the same host. Moreover, our data show that long-term tolerance induced by Foxp3(+) Treg-cell transfer is sustained by splenic Tr1 cells and functionally moves from the allograft to the spleen.

Biomodulatory role of ceramide in basic fibroblast growth factor-induced proliferation of cerebellar astrocytes in primary culture.

To evaluate the role of ceramide in glial growth, primary cultures of quiescent astrocytes from rat cerebellum were stimulated to proliferate by mitogenic doses of basic fibroblast growth factor (bFGF). Parallel to the bFGF mitogenic effect was a marked, and persistent, decrease in cellular ceramide levels. Both in vitro and in culture metabolic studies have led us to exclude both sphingomyelinase and ceramidase involvement in ceramide level variation. Instead, we found evidence of a functional connection between the decrease in ceramide levels and astrocyte proliferation. In fact, cell growth in bFGF-stimulated astrocytes was inhibited by exogenous ceramide and C2-ceramide, maximal inhibition being obtained at a ceramide concentration of 5-10 microM. Under the same conditions, the dihydroderivatives of ceramides were without effect. Following ceramide treatment, the phosphorylation of the MAP kinase isoforms ERK1/2, key components in bFGF-induced cell proliferation, was examined. The administration of antiproliferative doses of ceramide or C2-ceramide, but not of their dihydroderivatives, resulted in a significant inhibition of ERK1/2 activation. In conclusion, our data indicate that the prompt modulation of ceramide levels by bFGF is an early step associated with the signaling pathways responsible for the mitogenic activity of bFGF in astrocytes.

Processing and release of human proinsulin-cleavage products into culture media by different engineered non-endocrine cells: a specific assessment by capillary electrophoresis.

The aim of this study was to compare the metabolic pathway to mature insulin through the intermediate forms (32-33 split, 65-66 split, des31,32 and des64,65) in human or murine cells engineered for the release of wild-type human proinsulin and in a genetically mutated one, in the search for a new approach for an insulin-dependent diabetes mellitus cure by gene therapy. Primary human fibroblasts, myoblasts and stabilized cell lines (HepG2 and NIH3T3) were transduced either with a retroviral vector coding for wild-type proinsulin or for a genetically mutated one, carrying cleavage sites sensitive to furin. The pattern of all the proinsulin cleavage products released into the cell culture supernatants was analyzed by capillary electrophoresis. All the cells transduced with the wild-type gene released intact proinsulin. HepG2 released a considerable amount of 65-66 split and des64,65, while primary myoblasts released all the intermediate forms and a limited amount of mature insulin. All the cells transduced with a furin-sensitive proinsulin gene released a higher amount of mature insulin (23-59% conversion yield) than the cells expressing wild-type proinsulin, whereas the total insulin was nearly constant. Only primary cells released all the cleavage products. Screening a wide variety of non-endocrine cells has revealed a large difference in the processing and release of immature and mature insulin forms, pointing to human hepatic cells as the most efficacious. Capillary electrophoresis provided on-line and in a single run a complete overview of the proinsulin metabolic pathway in different cells.