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As well as being a significant source of environmental radiation exposure, α-particles are increasingly considered for use in targeted radiation therapy. A better understanding of α-particle induced damage at the DNA scale can be achieved by following their tracks in real-time in targeted living cells. Focused α-particle microbeams can facilitate this but, due to their low energy (up to a few MeV) and limited range, α-particles detection, delivery, and follow-up observations of radiation-induced damage remain difficult. In this study, we developed a thin Boron-doped Nano-Crystalline Diamond membrane that allows reliable single α-particles detection and single cell irradiation with negligible beam scattering. The radiation-induced responses of single 3 MeV α-particles delivered with focused microbeam are visualized in situ over thirty minutes after irradiation by the accumulation of the GFP-tagged RNF8 protein at DNA damaged sites., Competing Interests: The authors declare no competing financial interests.

The energy and specific energy absorbed in the main cell compartments (nucleus and cytoplasm) in typical radiobiology experiments are usually estimated by calculations as they are not accessible for a direct measurement. In most of the work, the cell geometry is modelled using the combination of simple mathematical volumes. We propose a method based on high resolution confocal imaging and ion beam analysis (IBA) in order to import realistic cell nuclei geometries in Monte-Carlo simulations and thus take into account the variety of different geometries encountered in a typical cell population. Seventy-six cell nuclei have been imaged using confocal microscopy and their chemical composition has been measured using IBA. A cellular phantom was created from these data using the ImageJ image analysis software and imported in the Geant4 Monte-Carlo simulation toolkit. Total energy and specific energy distributions in the 76 cell nuclei have been calculated for two types of irradiation protocols: a 3 MeV alpha particle microbeam used for targeted irradiation and a ²³⁹Pu alpha source used for large angle random irradiation. Qualitative images of the energy deposited along the particle tracks have been produced and show good agreement with images of DNA double strand break signalling proteins obtained experimentally. The methodology presented in this paper provides microdosimetric quantities calculated from realistic cellular volumes. It is based on open-source oriented software that is publicly available.

Deciphering the molecular basis of toxicology mechanism induced by nanoparticles (NPs) remains an essential challenge. Ion Beam Analysis (IBA) was applied in combination with Transmission Electron Microscopy and Confocal Microscopy to analyze human keratinocytes exposed to TiO(2)-NPs. Investigating chemical elemental distributions using IBA gives rise to a fine quantification of the TiO(2)-NPs uptake within a cell and to the determination of the intracellular chemical modifications after TiO(2)-NPs internalization. In addition, fluorescent dye-modified TiO(2)-NPs have been synthesized to allow their detection, precise quantification and tracking in vitro. The internalization of these TiO(2)-NPs altered the calcium homeostasis and induced a decrease in cell proliferation associated with an early keratinocyte differentiation, without any indication of cell death. Additionally, the relation between the surface chemistry of the TiO(2)-NPs and their in vitro toxicity is clearly established and emphasizes the importance of the calcium homeostasis alteration in response to the presence of TiO(2)-NPs.

Microbeam facilities provide a unique opportunity to investigate the effects of ionising radiation on living biological cells with a precise control of the delivered dose. This paper describes dosimetry calculations performed at the single-cell level in the microbeam irradiation facility available at the Centre d'Etudes Nucléaires de Bordeaux-Gradignan in France, using the object-oriented Geant4 Monte Carlo simulation toolkit. The cell geometry model is based on high-resolution three-dimensional voxelised phantoms of a human keratinocyte (HaCaT) cell line. Such phantoms are built from confocal microscopy imaging and from ion beam chemical elemental analysis. Results are presented for single-cell irradiation with 3 MeV incident alpha particles.

The ionic channels (particularly, K+ and Ca2+ channels) regulate, via the membrane potential, the ionic distribution into the vascular cells. Micro-particule induced X-ray emission (PIXE) analysis was applied to determine the ionic composition of vascular smooth muscle cells (VSMCs) and of vascular endothelial cells (VECs) in the placental human allantochorial vessels in a physiological medium (Hanks' solution) modified by the addition of a NO donor (sodium nitroprusside, SNP) and of a beta-adrenergic stimulator (isoproterenol, ISO). The addition of SNP or ISO induced no modification of the Na, K, Cl, P, S, Mg and Ca concentrations in VSMCs. In VECs, a same effect was observed except an increase of the Mg concentration with ISO. Theses results indicated a retroactive control (active feedback) of the internal ionic distribution by endothelial factors, ionic channels and exchangers.

Extracellular magnesium salts are known to interfere with ionic channels in the cellular membranes. The membrane potential, a regulator of vascular tone, is a function of the physiological activities of ionic channels (particularly, K+ and Ca2+ channels in these cells). These channels regulate the ionic distribution into these cells. Micro-Particule Induced X-ray Emission (PIXE) analysis was applied to determine the ionic composition of vascular smooth muscle cells (VSMC) and of vascular endothelial cells (VEC) in the placental human allantochorial vessels in a physiological medium (Hanks' solution) modified by the addition of 2 mM MgCl2 or 2 mM MgSO4 which block the calcium-sensitive K+ channels (K(Ca)), the ATP-sensitive K+ channels (K(ATP)) and the voltage-sensitive K+ (K(df)) and Ca2+ channels. In VSMC (media layer), the addition of MgCl2 induced no modification of the K, Cl, P, S and Ca concentrations but increased the Na and Mg concentrations and the addition of MgSO4 only significantly increased the Mg concentration, the other ion concentrations remaining constant. In endothelium (VEC), MgCl2 or MgSO4 addition implicated the same observations as in VSMC. These results confirmed the blockage of K(df), K(Ca), K(ATP) and Ca channels in VSMC and VEC by magnesium salts, the relationship between Mg2+ ions and internal Na and demonstrated the possible intervention of a Na+/Mg2+ exchanger.

The membrane potential, a regulator of vascular tone, is a function of the physiological activities of ionic channels (particularly, K+ and Ca2+ channels in these cells). These channels regulate the ionic distribution into these cells. Micro-particule induced X-ray emission (PIXE) analysis was applied to determine the ionic composition of vascular smooth muscle cells (VSMCs) and of vascular endothelial cells (VECs) in the placental human allantochorial vessels in a physiological medium (Hanks'solution) modified by the addition of a chemical stimulus: 5-hydroxytryptamine (5-HT), an activator of the voltage-sensitive Ca2+ channels. In VSMCs (media layer), the addition of 5-HT induced no modification of the Na, K, Cl, P, S and Ca concentrations but increased Mg concentration. In endothelium (VECs) 5-HT addition implicated an increase of the K, S, Ca concentrations, the concentration of the other ions remained constant. In VECs, Ca and K increase is due to open of L-type voltage-dependent Ca2+ channels and of K(Ca) channels. 5-HT induces also a secretion of endothelium hyperpolarizing factors which implicate decrease of [Ca2+]i in VSMCs opposite to a direct increase by 5-HT. Increase in [Mg2+]i may be due to activation of the Ca/Mg exchanger.

A 68-year-old cattle farmer from northern Senegal sought medical attention for tumefaction that had been progressing on the right foot and leg for 20 years. Physical examination of the right extremity revealed very firm tumefaction involving the foot and whole leg associated with numerous nodules. Bone radiographs and CT-scan of the foot and leg disclosed extensive osteolytic involvement. A specimen of squamous tissue from the top of nodules showed the presence of fumagoid cells characteristic of chromomycosis. Histologic examination after skin biopsy demonstrated fungal myocetoma. Due to the extent of involvement surgical and antifungal treatment was proposed but the patient refused to undergo surgery. Only one previous case of concurrent chromomycosis and mycetoma has been described. However the previous case involved actinomycetoma. The rarity of this combination of diseases despite their common contamination mode is due to different geographical distribution with mycetoma being found in the Sahelian region and chromomycosis in the humid equatorial region.