Sarcoma cell-specific radiation sensitization by titanate scrolled nanosheets: insights from physicochemical analysis and transcriptomic profiling.

This study aimed to explore the potential of metal oxides such as Titanate Scrolled Nanosheets (TNs) in improving the radiosensitivity of sarcoma cell lines. Enhancing the response of cancer cells to radiation therapy is crucial, and one promising approach involves utilizing metal oxide nanoparticles. We focused on the impact of exposing two human sarcoma cell lines to both TNs and ionizing radiation (IR). Our research was prompted by previous in vitro toxicity assessments, revealing a correlation between TNs' toxicity and alterations in intracellular calcium homeostasis. A hydrothermal process using titanium dioxide powder in an alkaline solution produced the TNs. Our study quantified the intracellular content of TNs and analyzed their impact on radiation-induced responses. This assessment encompassed PIXE analysis, cell proliferation, and transcriptomic analysis. We observed that sarcoma cells internalized TNs, causing alterations in intracellular calcium homeostasis. We also found that irradiation influence intracellular calcium levels. Transcriptomic analysis revealed marked disparities in the gene expression patterns between the two sarcoma cell lines, suggesting a potential cell-line-dependent nano-sensitization to IR. These results significantly advance our comprehension of the interplay between TNs, IR, and cancer cells, promising potential enhancement of radiation therapy efficiency.

Practical and technical key challenges in head and neck adaptive radiotherapy: The GORTEC point of view.

Anatomical variations occur during head and neck (H&N) radiotherapy (RT) treatment. These variations may result in underdosage to the target volume or overdosage to the organ at risk. Replanning during the treatment course can be triggered to overcome this issue. Due to technological, methodological and clinical evolutions, tools for adaptive RT (ART) are becoming increasingly sophisticated. The aim of this paper is to give an overview of the key steps of an H&N ART workflow and tools from the point of view of a group of French-speaking medical physicists and physicians (from GORTEC). Focuses are made on image registration, segmentation, estimation of the delivered dose of the day, workflow and quality assurance for an implementation of H&N offline and online ART. Practical recommendations are given to assist physicians and medical physicists in a clinical workflow.

Changes in intra-nuclear mechanics in response to DNA damaging agents revealed by time-domain Brillouin micro-spectroscopy.

How DNA damage and repair processes affect the biomechanical properties of the nucleus interior remains unknown. Here, an opto-acoustic microscope based on time-domain Brillouin spectroscopy (TDBS) was used to investigate the induced regulation of intra-nuclear mechanics. With this ultrafast pump-probe technique, coherent acoustic phonons were tracked along their propagation in the intra-nucleus nanostructure and the complex stiffness moduli and thicknesses were measured with an optical resolution. Osteosarcoma cells were exposed to methyl methanesulfonate (MMS) and the presence of DNA damage was tested using immunodetection targeted against damage signaling proteins. TDBS revealed that the intra-nuclear storage modulus decreased significantly upon exposure to MMS, as a result of the chromatin decondensation and reorganization that favors molecular diffusion within the organelle. When the damaging agent was removed and cells incubated for 2 h in the buffer solution before fixation the intra-nuclear reorganization led to an inverse evolution of the storage modulus, the nucleus stiffened. The same tendency was measured when DNA double-strand breaks were caused by cell exposure to ionizing radiation. TDBS microscopy also revealed changes in acoustic dissipation, another mechanical probe of the intra-nucleus organization at the nano-scale, and changes in nucleus thickness during exposure to MMS and after recovery.

Aberrations of plane-symmetrical mirror systems with freeform surfaces. Part I: generalized ray-tracing equations.

The aberrations of reflective optical systems with one plane of symmetry are investigated in the most general case, with freeform surfaces and possibly different locations of the tangential and sagittal object and image. In this first paper in a series of two, we establish generalized ray-tracing equations including transverse aberrations up to the third order in ray coordinates. The ray-tracing treatment allows us to overcome difficulties linked to the non-existence of a suitable astigmatic wavefront reference. The obtained expressions can describe multi-mirror systems and include all induced aberration terms. As an illustration, a simple freeform off-axis mirror is analyzed.

Aberrations of plane-symmetrical mirror systems with freeform surfaces. Part II: closed-form aberration formulas.

The aberrations of reflective optical systems with planar symmetry are investigated in the most general case, with freeform surfaces and possibly different locations of the tangential and sagittal object and image. In this second and last paper, closed-form expressions are derived for the aberrations created by an individual mirror. We study two-mirror off-axis telescopes and establish a new family of designs that we show is simultaneously free of constant coma, linear astigmatism, and quadratic distortions (including smile and keystone). Analytical expressions for the intrinsic aberrations of an astigmatic beam are also derived.

Use of metrics to quantify IMRT and VMAT treatment plan complexity: A systematic review and perspectives.

PURPOSE: Fixed-field intensity modulated radiation therapy (FF-IMRT) or volumetric modulated arc therapy (VMAT) beams complexity is due to fluence fluctuation. Pre-treatment Quality Assurance (PTQA) failure could be linked to it. Several plan complexity metrics (PCM) have been published to quantify this complexity but in a heterogeneous formalism. This review proposes to gather different PCM and to discuss their eventual PTQA failure identifier abilities. METHODS AND MATERIALS: A systematic literature search and outcome extraction from MEDLINE/PubMed (National Center for Biotechnology Information, NCBI) was performed. First, a list and a synthesis of available PCM is made in a homogeneous formalism. Second, main results relying on the link between PCM and PTQA results but also on other uses are listed. RESULTS: A total of 163 studies were identified and n = 19 were selected after inclusion and exclusion criteria application. Difference is made between fluence and degree of freedom (DOF)-based PCM. Results about the PCM potential as PTQA failure identifier are described and synthesized. Others uses are also found in quality, big data, machine learning and audit procedure. CONCLUSIONS: A state of the art is made thanks to this homogeneous PCM classification. For now, PCM should be seen as a planning procedure quality indicator although PTQA failure identifier results are mitigated. However limited clinical use seems possible for some cases. Yet, addressing the general PTQA failure prediction case could be possible with the big data or machine learning help.

Low-density lipoprotein receptor-deficient hepatocytes differentiated from induced pluripotent stem cells allow familial hypercholesterolemia modeling, CRISPR/Cas-mediated genetic correction, and productive hepatitis C virus infection.

BACKGROUND: Familial hypercholesterolemia type IIA (FH) is due to mutations in the low-density lipoprotein receptor (LDLR) resulting in elevated levels of low-density lipoprotein cholesterol (LDL-c) in plasma and in premature cardiovascular diseases. As hepatocytes are the only cells capable of metabolizing cholesterol, they are therefore the target cells for cell/gene therapy approaches in the treatment of lipid metabolism disorders. Furthermore, the LDLR has been reported to be involved in hepatitis C virus (HCV) entry into hepatocytes; however, its role in the virus infection cycle is still disputed. METHODS: We generated induced pluripotent stem cells (iPSCs) from a homozygous LDLR-null FH-patient (FH-iPSCs). We constructed a correction cassette bearing LDLR cDNA under the control of human hepatic apolipoprotein A2 promoter that targets the adeno-associated virus integration site AAVS1. We differentiated both FH-iPSCs and corrected FH-iPSCs (corr-FH-iPSCs) into hepatocytes to study statin-mediated regulation of genes involved in cholesterol metabolism. Upon HCV particle inoculation, viral replication and production were quantified in these cells. RESULTS: We showed that FH-iPSCs displayed the disease phenotype. Using homologous recombination mediated by the CRISPR/Cas9 system, FH-iPSCs were genetically corrected by the targeted integration of a correction cassette at the AAVS1 locus. Both FH-iPSCs and corr-FH-iPSCs were then differentiated into functional polarized hepatocytes using a stepwise differentiation approach (FH-iHeps and corr-FH-iHeps). The correct insertion and expression of the correction cassette resulted in restoration of LDLR expression and function (LDL-c uptake) in corr-FH-iHeps. We next demonstrated that pravastatin treatment increased the expression of genes involved in cholesterol metabolism in both cell models. Moreover, LDLR expression and function were also enhanced in corr-FH-iHeps after pravastatin treatment. Finally, we demonstrated that both FH-iHeps and corr-FH-iHeps were as permissive to viral infection as primary human hepatocytes but that virus production in FH-iHeps was significantly decreased compared to corr-FH-iHeps, suggesting a role of the LDLR in HCV morphogenesis. CONCLUSIONS: Our work provides the first LDLR-null FH cell model and its corrected counterpart to study the regulation of cholesterol metabolism and host determinants of HCV life cycle, and a platform to screen drugs for treating dyslipidemia and HCV infection.

Photochemistry on the Space Station-Aptamer Resistance to Space Conditions: Particles Exposure from Irradiation Facilities and Real Exposure Outside the International Space Station.

Some microarray-based instruments that use bioaffinity receptors such as antibodies or aptamers are under development to detect signatures of past or present life on planetary bodies. Studying the resistance of such instruments against space constraints and cosmic rays in particular is a prerequisite. We used several ground-based facilities to study the resistance of aptamers to various types of particles (protons, electrons, neutrons, and carbon ions) at different energies and fluences. We also tested the resistance of aptamers during the EXPOSE-R2 mission outside the International Space Station (ISS). The accumulated dose measured after the 588 days of this mission (220 mGy) corresponds to the accumulated dose that can be expected during a mission to Mars. We found that the recognition ability of fluorescently labeled aptamers was not significantly affected during short-term exposure experiments taking into account only one type of radiation at a time. However, we demonstrated that the same fluorescent dye was significantly affected by temperature variations (-21°C to +58°C) and storage throughout the entirety of the ISS experiment (60% of signal loss). This induced a large variability of aptamer signal in our analysis. However, we found that >50% of aptamers were still functional after the whole EXPOSE-R2 mission. We conclude that aptamer-based instruments are well suited for in situ analysis on planetary bodies, but the detection step requires additional investigations.

High performance modelling of the transport of energetic particles for photon radiotherapy.

This work consists of the validation of a new Grid Based Boltzmann Solver (GBBS) conceived for the description of the transport and energy deposition by energetic particles for radiotherapy purposes. The entropic closure and a compact mathematical formulation allow our code (M1) to calculate the delivered dose with an accuracy comparable to the Monte-Carlo (MC) codes with a computational time that is reduced to the order of few minutes without any special processing power requirement. A validation protocol with heterogeneity inserts has been defined for different photon sources. The comparison with the MC calculated depth-dose curves and transverse profiles of the beam at different depths shows an excellent accuracy of the M1 model.

Transplantation of hESC-derived hepatocytes protects mice from liver injury.

BACKGROUND: Hepatic cell therapy has become a viable alternative to liver transplantation for life-threatening liver diseases. However, the supply of human hepatocytes is limited due to the shortage of suitable donor organs required to isolate high-quality cells. Human pluripotent stem cells reflect a potential renewable source for generating functional hepatocytes. However, most differentiation protocols use undefined matrices or factors of animal origin; as such, the resulting hepatocytes are not Good Manufacturing Practice compliant. Moreover, the preclinical studies employed to assess safety and function of human embryonic stem cell (hESC)-derived hepatocytes are generally limited to immunodeficient mice. In the present study, we evaluate the generation of hepatocytes under defined conditions using a European hESC line (VAL9) which was derived under animal-free conditions. The function capacity of VAL9-derived hepatocytes was assessed by transplantation into mice with acetaminophen-induced acute liver failure, a clinically relevant model. METHODS: We developed a protocol that successfully differentiates hESCs into bipotent hepatic progenitors under defined conditions, without the use of chromatin modifiers such as dimethyl sulphoxide. These progenitors can be cryopreserved and are able to generate both committed precursors of cholangiocytes and neonate-like hepatocytes. RESULTS: Thirty days post-differentiation, hESCs expressed hepatocyte-specific markers such as asialoglycoprotein receptor and hepatic nuclear factors including HNF4α. The cells exhibited properties of mature hepatocytes such as urea secretion and UGT1A1 and cytochrome P450 activities. When transplanted into mice with acetaminophen-induced acute liver failure, a model of liver damage, the VAL9-derived hepatocytes efficiently engrafted and proliferated, repopulating up to 10 % of the liver. In these transplanted livers, we observed a significant decrease of liver transaminases and found no evidence of tumourigenicity. Thus, VAL9-derived hepatocytes were able to rescue hepatic function in acetaminophen-treated animals. CONCLUSIONS: Our study reveals an efficient protocol for differentiating VAL9 hESCs to neonatal hepatocytes which are then able to repopulate livers in vivo without tumour induction. The human hepatocytes are able to rescue liver function in mice with acetaminophen-induced acute toxicity. These results provide proof-of-concept that replacement therapies using hESC-derived hepatocytes are effective for treating liver diseases.

Dose calculation accuracy of different image value to density tables for cone-beam CT planning in head & neck and pelvic localizations.

PURPOSE: We aimed to identify the most accurate combination of phantom and protocol for image value to density table (IVDT) on volume-modulated arc therapy (VMAT) dose calculation based on kV-Cone-beam CT imaging, for head and neck (H&N) and pelvic localizations. METHODS: Three phantoms (Catphan(®)600, CIRS(®)062M (inner phantom for head and outer phantom for body), and TomoTherapy(®) "Cheese" phantom) were used to create IVDT curves of CBCT systems with two different CBCT protocols (Standard-dose Head and Standard Pelvis). Hounsfield Unit (HU) time stability and repeatability for a single On-Board-Imager (OBI) and compatibility of two distinct devices were assessed with Catphan(®)600. Images from the anthropomorphic phantom CIRS ATOM(®) for both CT and CBCT modalities were used for VMAT dose calculation from different IVDT curves. Dosimetric indices from CT and CBCT imaging were compared. RESULTS: IVDT curves from CBCT images were highly different depending on phantom used (up to 1000 HU for high densities) and protocol applied (up to 200 HU for high densities). HU time stability was verified over seven weeks. A maximum difference of 3% on the dose calculation indices studied was found between CT and CBCT VMAT dose calculation across the two localizations using appropriate IVDT curves. One IVDT curve per localization can be established with a bi-monthly verification of IVDT-CBCT. CONCLUSIONS: The IVDT-CBCTCIRS-Head phantom with the Standard-dose Head protocol was the most accurate combination for dose calculation on H&N CBCT images. For pelvic localizations, the IVDT-CBCTCheese established with the Standard Pelvis protocol provided the best accuracy.

Operating wavelengths optimization for a spaceborne lidar measuring atmospheric CO2.

The Advanced Space Carbon and Climate Observation of Planet Earth (A-SCOPE) mission, a candidate for the next generation of European Space Agency Earth Explorer Core Missions, aims at measuring CO(2) concentration from space with an integrated path differential absorption (IPDA) lidar. We report the optimization of the lidar instrument operating wavelengths, building on two performance models developed to assess measurement random errors from the instrument, as well as knowledge errors on geophysical and spectral parameters required for the measurement processing. A promising approach to decrease sensitivity to water vapor errors by 1 order of magnitude is reported and illustrated. The presented methods are applicable for any airborne or spaceborne IPDA lidar.

Infrared specular reflection spectroscopy of rough metallic substrates.

A study of the infrared reflectance of rough metallic surfaces is presented. We show that one reflectance measurement, made in the specular direction under specific conditions, allows the accurate calculation of the shape of the roughness histogram. As a theoretical background, we use modified expressions from Kirchhoff theory for surface scattering. To illustrate our method, we present experimental results obtained with surfaces having a multimode histogram, which means having several different populations with distinct peaks. For these surfaces, we observe oscillations in the regularly decreasing reflectance (with decreasing wavelength) that are created by a partial interference phenomenon between the peaks. To our knowledge, this effect is presented for the first time in the literature. Our study demonstrates that some very useful information can be obtained in the infrared spectrum of metallic substrates, although they do not have any absorption band. We hope that our results help further the understanding of complex spectral data obtained on such scattering substrates covered with organic films.

Voxeldoes: a computer program for 3-D dose calculation in therapeutic nuclear medicine.

A computer program, VoxelDose, was developed to calculate patient specific 3-D-dose maps at the voxel level. The 3-D dose map is derived in three steps: (i) The SPECT acquisitions are reconstructed using a filtered back projection method, with correction for attenuation and scatter; (ii) the 3-D cumulated activity map is generated by integrating the SPECT data; and (iii) a 3-D dose map is computed by convolution (using the Fourier Transform) of the cumulated activity map and corresponding MIRD voxel S values. To validate the VoxelDose software, a Liqui-Phil abdominal phantom with four simulated organ inserts and one spherical tumor (radius 4.2 cm) was filled with known activity concentrations of 111In. Four cylindrical calibration tubes (from 3.7 to 102 kBq/mL) were placed on the phantom. Thermoluminescent mini-dosimeters (mini-TLDs) were positioned on the surface of the organ inserts. Percent differences between the known and measured activity concentrations were determined to be 12.1 (tumor), 1.8 (spleen), 1.4, 8.1 (right and left kidneys), and 38.2% (liver), leading to percent differences between the calculated and TLD measured doses of 41, 16, 3, 5, and 62%. Large differences between the measured and calculated dose in the tumor and the liver may be attributed to several reasons, such as the difficulty in precisely associating the position of the TLD to a voxel and limits of the quantification method (mainly the scatter correction and partial volume effect). Further investigations should be performed to better understand the impact of each effect on the results and to improve absolute quantification. For all other organs, activity concentration measurements and dose calculations agree well with the known activity concentrations.