Two-photon intravital imaging of lungs during anthrax infection reveals long-lasting macrophage-dendritic cell contacts.

International audience: Dynamics of the lung immune system at a microscopic level are largely unknown because of inefficient methods to rid chest motion during image acquisition. In this study, we developed an improved intravital method for two-photon lung imaging uniquely based on a posteriori parenchymal tissue motion correction. We took advantage of the alveolar collagen pattern given by second harmonic generation signal as a reference for frame registration. We describe here for the first time a detailed dynamic account of two major lung immune cell populations, alveolar macrophages and CD11b-positive dendritic cells, during homeostasis and infection by spores of Bacillus anthracis, the agent of anthrax. We show that after alveolar macrophages capture spores, CD11b-positive dendritic cells come in prolonged contact with infected macrophages. Dendritic cells are known to carry spores to the draining lymph nodes and elicit the immune response in pulmonary anthrax. The intimate and long-lasting contacts between these two lines of defense may therefore coordinate immune responses in the lung through an immunological synapse-like process

Intracerebral delivery of Carboplatin in combination with either 6 MV Photons or monoenergetic synchrotron X-rays are equally efficacious for treatment of the F98 rat glioma.

International audienceABSTRACT: BACKGROUND: The purpose of the present study was to compare side-by-side the therapeutic efficacy of a 6-day infusion of carboplatin, followed by X-irradiation with either 6 MV photons or synchrotron X-rays, tuned above the K-edge of Pt, for treatment of F98 glioma bearing rats. METHODS: Carboplatin was administered intracerebrally (i.c.) to F98 glioma bearing rats over 6 days using AlzetTM osmotic pumps starting 7 days after tumor implantation. Radiotherapy was delivered in a single 15 Gy fraction on day 14 using a conventional 6 MV linear accelerator (LINAC) or 78.8 keV synchrotron X-rays. RESULTS: Untreated control animals had a median survival time (MeST) of 33 days. Animals that received either carboplatin alone or irradiation alone with either 78.8 keV or 6 MV had a MeSTs 38 and 33 days, respectively. Animals that received carboplatin in combination with X-irradiation had a MeST of > 180 days with a 55% cure rate, irrespective of whether they were irradiated with either 78.8 KeV synchrotron X-rays or 6MV photons. CONCLUSIONS: These studies have conclusively demonstrated the equivalency of i.c. delivery of carboplatin in combination with X-irradiation with either 6 MV photons or synchrotron X-rays

Intracerebral delivery of 5-iodo-2'-deoxyuridine in combination with synchrotron stereotactic radiation for the therapy of the F98 glioma.

International audienceIodine-enhanced synchrotron stereotactic radiotherapy takes advantage of the radiation dose-enhancement produced by high-Z elements when irradiated with mono-energetic beams of synchrotron X-rays. In this study it has been investigated whether therapeutic efficacy could be improved using a thymidine analogue, 5-iodo-2'-deoxyuridine (IUdR), as a radiosentizing agent. IUdR was administered intracerebrally over six days to F98 glioma-bearing rats using Alzet osmotic pumps, beginning seven days after tumor implantation. On the 14th day, a single 15 Gy dose of 50 keV synchrotron X-rays was delivered to the brain. Animals were followed until the time of death and the primary endpoints of this study were the mean and median survival times. The median survival times for irradiation alone, chemotherapy alone or their combination were 44, 32 and 46 days, respectively, compared with 24 days for untreated controls. Each treatment alone significantly increased the rats' survival in comparison with the untreated group. Their combination did not, however, significantly improve survival compared with that of X-irradiation alone or chemotherapy alone. Further studies are required to understand why the combination of chemoradiotherapy was no more effective than X-irradiation alone

IntrAnat Electrodes: A Free Database and Visualization Software for Intracranial Electroencephalographic Data Processed for Case and Group Studies

In some cases of pharmaco-resistant and focal epilepsies, intracranial recordings performed epidurally (electrocorticography, ECoG) and/or in depth (stereoelectroencephalography, SEEG) can be required to locate the seizure onset zone and the eloquent cortex before surgical resection. In SEEG, each electrode contact records brain’s electrical activity in a spherical volume of 3 mm diameter approximately. The spatial coverage is around 1% of the brain and differs between patients because the implantation of electrodes is tailored for each case. Group studies thus need a large number of patients to reach a large spatial sampling, which can be achieved more easily using a multicentric approach such as implemented in our F-TRACT project (f-tract.eu). To facilitate group studies, we developed a software—IntrAnat Electrodes—that allows to perform virtual electrode implantation in patients’ neuroanatomy and to overlay results of epileptic and functional mapping, as well as resection masks from the surgery. IntrAnat Electrodes is based on a patient database providing multiple search criteria to highlight various group features. For each patient, the anatomical processing is based on a series of software publicly available. Imaging modalities (Positron Emission Tomography (PET), anatomical MRI pre-implantation, post-implantation and post-resection, functional MRI, diffusion MRI, Computed Tomography (CT) with electrodes) are coregistered. The 3D T1 pre-implantation MRI gray/white matter is segmented and spatially normalized to obtain a series of cortical parcels using different neuroanatomical atlases. On post-implantation images, the user can position 3D models of electrodes defined by their geometry. Each electrode contact is then labeled according to its position in the anatomical atlases, to the class of tissue (gray or white matter, cerebro-spinal fluid) and to its presence inside or outside the resection mask. Users can add more functionally informed labels on contact, such as clinical responses after electrical stimulation, cortico-cortical evoked potentials, gamma band activity during cognitive tasks or epileptogenicity. IntrAnat Electrodes software thus provides a means to visualize multimodal data. The contact labels allow to search for patients in the database according to multiple criteria representing almost all available data, which is to our knowledge unique in current SEEG software. IntrAnat Electrodes will be available in the forthcoming release of BrainVisa software and tutorials can be found on the F-TRACT webpage

Probabilistic functional tractography of the human cortex revisited

In patients with pharmaco-resistant focal epilepsies investigated with intracranial electroencephalography (iEEG), direct electrical stimulations of a cortical region induce cortico-cortical evoked potentials (CCEP) in distant cerebral cortex, which properties can be used to infer large scale brain connectivity. In 2013, we proposed a new probabilistic functional tractography methodology to study human brain connectivity. We have now been revisiting this method in the F-TRACT project (f-tract.eu) by developing a large multicenter CCEP database of several thousand stimulation runs performed in several hundred patients, and associated processing tools to create a probabilistic atlas of human cortico-cortical connections. Here, we wish to present a snapshot of the methods and data of F-TRACT using a pool of 213 epilepsy patients, all studied by stereo-encephalography with intracerebral depth electrodes. The CCEPs were processed using an automated pipeline with the following consecutive steps: detection of each stimulation run from stimulation artifacts in raw intracranial EEG (iEEG) files, bad channels detection with a machine learning approach, model-based stimulation artifact correction, robust averaging over stimulation pulses. Effective connectivity between the stimulated and recording areas is then inferred from the properties of the first CCEP component, i.e. onset and peak latency, amplitude, duration and integral of the significant part. Finally, group statistics of CCEP features are implemented for each brain parcel explored by iEEG electrodes. The localization (coordinates, white/gray matter relative positioning) of electrode contacts were obtained from imaging data (anatomical MRI or CT scans before and after electrodes implantation). The iEEG contacts were repositioned in different brain parcellations from the segmentation of patients' anatomical MRI or from templates in the MNI coordinate system. The F-TRACT database using the first pool of 213 patients provided connectivity probability values for 95% of possible intrahemispheric and 56% of interhemispheric connections and CCEP features for 78% of intrahemisheric and 14% of interhemispheric connections. In this report, we show some examples of anatomo-functional connectivity matrices, and associated directional maps. We also indicate how CCEP features, especially latencies, are related to spatial distances, and allow estimating the velocity distribution of neuronal signals at a large scale. Finally, we describe the impact on the estimated connectivity of the stimulation charge and of the contact localization according to the white or gray matter. The most relevant maps for the scientific community are available for download on f-tract. eu (David et al., 2017) and will be regularly updated during the following months with the addition of more data in the F-TRACT database. This will provide an unprecedented knowledge on the dynamical properties of large fiber tracts in human.Peer reviewe

Brain tumors and synchrotron radiation : new methods for minibeams radiation therapy and treatment follow-up by functional imaging.

En 2006 Dilmanian et al. ont proposé une méthode d'irradiation par rayonnement synchrotron innovante appelée minifaisceaux. L'irradiation de tumeur par minifaisceaux monochromatiques consiste en un motif fractionné spatialement de faisceaux de rayons X submillimétriques produits par un synchrotron. Afin d'obtenir une dose homogène dans le volume cible, deux incidences orthogonales sont entrecroisées. Le tissu sain environnant ne subit que l'irradiation fractionnée, entre les faisceaux la dose n'est dûe qu'au diffusé et l'énergie déposée y est donc 10 à 15 fois inférieure à celle déposée sur les axes des faisceaux. Cela permet un effet protecteur des tissus sains tout en distribuant de fortes doses à la tumeur. Cette thèse porte sur le développement de la méthode expérimentale des minifaisceaux monochromatiques, ce qui comprend le contrôle de la géométrie d'irradiation, la dosimétrie expérimentale et l'étude Monte Carlo correspondante. Afin d'évaluer son efficacité, des études précliniques ont été réalisées sur un modèle de tumeur cérébrale implantée chez le rat (F98). Un suivi de traitement est réalisé par de l'imagerie anatomique et fonctionnelle afin d'évaluer son efficacité. L'imagerie de perfusion cérébrale (menant aux volumes et débits sanguin cérébrales, au temps de transit moyen) est d'après la littérature un moyen efficace de pronostique du résultat du traitement. Les paramètres clés de la vascularisation cérébrale sont principalement étudiés par imagerie IRM, du fait de l'innocuité de ce type d'imagerie. La Synchrotron Radiation Computed Tomography (SRCT) est une modalité d'imagerie dont les performances sont proches des limites théoriques dans l'obtention de mesures absolues des concentrations d'agent de contraste et peut être utilisé en tant que gold-standard. Les modèles pharmacocinétiques utilisés nécessitent comme paramètre d'entrée des concentrations d'agent de contraste en fonction du temps. La relation entre le signal obtenu par IRM et la concentration d'agent de contraste est très complexe et difficilement quantitative. Une comparaison des mesures de perfusion effectuées par IRM et par SRCT a été effectuée afin de calibrer les mesures IRM.An innovative method of synchrotron radiation therapy, called minibeams, was proposed by A. Dilmanian et al. in 2006. Minibeams consists in tumor irradiation with monochromatic submillimetric x-ray beams spatially fractionated produced by a synchrotron source. To obtain a homogeneous dose in the target volume, an interleaving is realized using two orthogonal incidences. Adjacent healthy tissue is only partially irradiated by minibeams, the areas between the beams only receive scattered radiation and therefore the energy deposited is 10 to 15 times lower than on one minibeam axis, leading to a sparing effect of healthy tissue even when a high dose is deposited in the target volume. The thesis project is the development of this experimental method of monochromatic minibeams, which involves the control of the irradiation geometry, the control of dosimetry and its modeling by Monte Carlo simulations. To evaluate the method, preclinical experiments on models of brain tumors implanted in rats (F98) are performed. Follow-up by anatomical and functional imaging is carried out to evaluate the effectiveness of the treatment. Functional imaging of cerebral perfusion (volume and cerebral blood flow, mean transit time of heavy elements) appears to be associated in the literature as a relevant method for monitoring prognostic. The key parameters of the cerebral vasculature are mainly studied in magnetic resonance imaging (MRI), because of the harmlessness of this imaging modality. The relation between MRI signal and contrast agent concentration is very complex and no quantitative relationship is well known. Synchrotron Radiation Computed Tomography (SRCT) is an imaging modality with performances to measure absolute contrast agent concentration very close to the theoretical limits and can be used as gold-standard. The used pharmacokinetic models need as input parameters a contrast agent concentration versus time. A comparison of perfusion measurements between MRI and SRCT has been done in order to calibrate MRI measurements

Tumeurs cérébrales et rayonnement Synchrotron. Développement méthodologique pour la radiothérapie par minifaisceaux et suivi du traitement par imagerie fonctionnelle

An innovative method of synchrotron radiation therapy, called minibeams, was proposed by A. Dilmanian et al. in 2006. Minibeams consists in tumor irradiation with monochromatic submillimetric x-ray beams spatially fractionated produced by a synchrotron source. To obtain a homogeneous dose in the target volume, an interleaving is realized using two orthogonal incidences. Adjacent healthy tissue is only partially irradiated by minibeams, the areas between the beams only receive scattered radiation and therefore the energy deposited is 10 to 15 times lower than on one minibeam axis, leading to a sparing effect of healthy tissue even when a high dose is deposited in the target volume. The thesis project is the development of this experimental method of monochromatic minibeams, which involves the control of the irradiation geometry, the control of dosimetry and its modeling by Monte Carlo simulations. To evaluate the method, preclinical experiments on models of brain tumors implanted in rats (F98) are performed. Follow-up by anatomical and functional imaging is carried out to evaluate the effectiveness of the treatment. Functional imaging of cerebral perfusion (volume and cerebral blood flow, mean transit time of heavy elements) appears to be associated in the literature as a relevant method for monitoring prognostic. The key parameters of the cerebral vasculature are mainly studied in magnetic resonance imaging (MRI), because of the harmlessness of this imaging modality. The relation between MRI signal and contrast agent concentration is very complex and no quantitative relationship is well known. Synchrotron Radiation Computed Tomography (SRCT) is an imaging modality with performances to measure absolute contrast agent concentration very close to the theoretical limits and can be used as gold-standard. The used pharmacokinetic models need as input parameters a contrast agent concentration versus time. A comparison of perfusion measurements between MRI and SRCT has been done in order to calibrate MRI measurements.En 2006 Dilmanian et al. ont proposé une méthode d'irradiation par rayonnement synchrotron innovante appelée minifaisceaux. L'irradiation de tumeur par minifaisceaux monochromatiques consiste en un motif fractionné spatialement de faisceaux de rayons X submillimétriques produits par un synchrotron. Afin d'obtenir une dose homogène dans le volume cible, deux incidences orthogonales sont entrecroisées. Le tissu sain environnant ne subit que l'irradiation fractionnée, entre les faisceaux la dose n'est dûe qu'au diffusé et l'énergie déposée y est donc 10 à 15 fois inférieure à celle déposée sur les axes des faisceaux. Cela permet un effet protecteur des tissus sains tout en distribuant de fortes doses à la tumeur. Cette thèse porte sur le développement de la méthode expérimentale des minifaisceaux monochromatiques, ce qui comprend le contrôle de la géométrie d'irradiation, la dosimétrie expérimentale et l'étude Monte Carlo correspondante. Afin d'évaluer son efficacité, des études précliniques ont été réalisées sur un modèle de tumeur cérébrale implantée chez le rat (F98). Un suivi de traitement est réalisé par de l'imagerie anatomique et fonctionnelle afin d'évaluer son efficacité. L'imagerie de perfusion cérébrale (menant aux volumes et débits sanguin cérébrales, au temps de transit moyen) est d'après la littérature un moyen efficace de pronostique du résultat du traitement. Les paramètres clés de la vascularisation cérébrale sont principalement étudiés par imagerie IRM, du fait de l'innocuité de ce type d'imagerie. La Synchrotron Radiation Computed Tomography (SRCT) est une modalité d'imagerie dont les performances sont proches des limites théoriques dans l'obtention de mesures absolues des concentrations d'agent de contraste et peut être utilisé en tant que gold-standard. Les modèles pharmacocinétiques utilisés nécessitent comme paramètre d'entrée des concentrations d'agent de contraste en fonction du temps. La relation entre le signal obtenu par IRM et la concentration d'agent de contraste est très complexe et difficilement quantitative. Une comparaison des mesures de perfusion effectuées par IRM et par SRCT a été effectuée afin de calibrer les mesures IRM

Tumeurs cérébrales et rayonnement Synchrotron. Développement méthodologique pour la radiothérapie par minifaisceaux et suivi du traitement par imagerie fonctionnelle

An innovative method of synchrotron radiation therapy, called minibeams, was proposed by A. Dilmanian et al. in 2006. Minibeams consists in tumor irradiation with monochromatic submillimetric x-ray beams spatially fractionated produced by a synchrotron source. To obtain a homogeneous dose in the target volume, an interleaving is realized using two orthogonal incidences. Adjacent healthy tissue is only partially irradiated by minibeams, the areas between the beams only receive scattered radiation and therefore the energy deposited is 10 to 15 times lower than on one minibeam axis, leading to a sparing effect of healthy tissue even when a high dose is deposited in the target volume. The thesis project is the development of this experimental method of monochromatic minibeams, which involves the control of the irradiation geometry, the control of dosimetry and its modeling by Monte Carlo simulations. To evaluate the method, preclinical experiments on models of brain tumors implanted in rats (F98) are performed. Follow-up by anatomical and functional imaging is carried out to evaluate the effectiveness of the treatment. Functional imaging of cerebral perfusion (volume and cerebral blood flow, mean transit time of heavy elements) appears to be associated in the literature as a relevant method for monitoring prognostic. The key parameters of the cerebral vasculature are mainly studied in magnetic resonance imaging (MRI), because of the harmlessness of this imaging modality. The relation between MRI signal and contrast agent concentration is very complex and no quantitative relationship is well known. Synchrotron Radiation Computed Tomography (SRCT) is an imaging modality with performances to measure absolute contrast agent concentration very close to the theoretical limits and can be used as gold-standard. The used pharmacokinetic models need as input parameters a contrast agent concentration versus time. A comparison of perfusion measurements between MRI and SRCT has been done in order to calibrate MRI measurements.En 2006 Dilmanian et al. ont proposé une méthode d'irradiation par rayonnement synchrotron innovante appelée minifaisceaux. L'irradiation de tumeur par minifaisceaux monochromatiques consiste en un motif fractionné spatialement de faisceaux de rayons X submillimétriques produits par un synchrotron. Afin d'obtenir une dose homogène dans le volume cible, deux incidences orthogonales sont entrecroisées. Le tissu sain environnant ne subit que l'irradiation fractionnée, entre les faisceaux la dose n'est dûe qu'au diffusé et l'énergie déposée y est donc 10 à 15 fois inférieure à celle déposée sur les axes des faisceaux. Cela permet un effet protecteur des tissus sains tout en distribuant de fortes doses à la tumeur. Cette thèse porte sur le développement de la méthode expérimentale des minifaisceaux monochromatiques, ce qui comprend le contrôle de la géométrie d'irradiation, la dosimétrie expérimentale et l'étude Monte Carlo correspondante. Afin d'évaluer son efficacité, des études précliniques ont été réalisées sur un modèle de tumeur cérébrale implantée chez le rat (F98). Un suivi de traitement est réalisé par de l'imagerie anatomique et fonctionnelle afin d'évaluer son efficacité. L'imagerie de perfusion cérébrale (menant aux volumes et débits sanguin cérébrales, au temps de transit moyen) est d'après la littérature un moyen efficace de pronostique du résultat du traitement. Les paramètres clés de la vascularisation cérébrale sont principalement étudiés par imagerie IRM, du fait de l'innocuité de ce type d'imagerie. La Synchrotron Radiation Computed Tomography (SRCT) est une modalité d'imagerie dont les performances sont proches des limites théoriques dans l'obtention de mesures absolues des concentrations d'agent de contraste et peut être utilisé en tant que gold-standard. Les modèles pharmacocinétiques utilisés nécessitent comme paramètre d'entrée des concentrations d'agent de contraste en fonction du temps. La relation entre le signal obtenu par IRM et la concentration d'agent de contraste est très complexe et difficilement quantitative. Une comparaison des mesures de perfusion effectuées par IRM et par SRCT a été effectuée afin de calibrer les mesures IRM