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International audience; Thanks to the recent development of sensors and Internet of Things (IoT), it is now common to use mobile application to monitor health status. These applications rely on sensors embedded in the smartphones that measure several physical quantities such as acceleration or angular velocity. However, these data are private information that can be used to infer sensitive attributes. This paper presents a new approach to anonymize the motion sensor data, preventing the re-identification of the user based on a selection of handcrafted features extracted from the distribution of zeros of the Shot-Time Fourier Transform (STFT). This work is motivated by recent works which highlight the importance of the zeros of the STFT Flandrin (IEEE Processing Letters 22:2137-2141, 1) and link them in the case of white noise to Gaussian Analytical Functions (GAF) Bardenet et al. (Applied and Computational Harmonic Analysis 48:682-705, 2) where the distribution of their zeros is formally described. The proposed approach is compared with an extension of an earlier work based on filtering in the time-frequency plane and doing the classification task based on convolutional neural networks, for which we improved the evaluation method and investigated the benefits of gyroscopic sensor’s data. An extensive comparison is performed on a first public dataset to assess the accuracy of activity recognition and user re-identification. We showed not only that the proposed method gives better results in term of activity/identity recognition trade-off compared with the state of the art but also that it can be generalized to other datasets.

Coats plus (CP) syndrome is an inherited autosomal recessive condition that results from mutations in the conserved telomere maintenance component 1 gene (CTC1). The CTC1 protein functions as a part of the CST protein complex, a protein heterotrimer consisting of CTC1–STN1–TEN1 which promotes telomere DNA synthesis and inhibits telomerase-mediated telomere elongation. However, it is unclear how CTC1 mutations may have an effect on telomere structure and function. For that purpose, we established the very first induced pluripotent stem cell lines (iPSCs) from a compound heterozygous patient with CP carrying deleterious mutations in both alleles of CTC1. Telomere dysfunction and chromosomal instability were assessed in both circulating lymphocytes and iPSCs from the patient and from healthy controls of similar age. The circulating lymphocytes and iPSCs from the CP patient were characterized by their higher telomere length heterogeneity and telomere aberrations compared to those in control cells from healthy donors. Moreover, in contrast to iPSCs from healthy controls, the high levels of telomerase were associated with activation of the alternative lengthening of telomere (ALT) pathway in CP-iPSCs. This was accompanied by inappropriate activation of the DNA repair proteins γH2AX, 53BP1, and ATM, as well as with accumulation of DNA damage, micronuclei, and anaphase bridges. CP-iPSCs presented features of cellular senescence and increased radiation sensitivity. Clonal dicentric chromosomes were identified only in CP-iPSCs after exposure to radiation, thus mirroring the role of telomere dysfunction in their formation. These data demonstrate that iPSCs derived from CP patients can be used as a model system for molecular studies of the CP syndrome and underscores the complexity of telomere dysfunction associated with the defect of DNA repair machinery in the CP syndrome.

OBJECTIVE: To test the hypothesis that telomere shortening and/or loss are risk factors for infertility.DESIGN: Retrospective analysis of the telomere status in patients with infertility using conventional cytogenetic data collected prospectively.SETTING: Academic centers.PATIENT(S): Cytogenetic slides with cultured peripheral lymphocytes from 50 patients undergoing fertility treatment and 150 healthy donors, including 100 donors matched for age.INTERVENTION(S): Cytogenetic slides were used to detect chromosomal and telomere aberrations.MAIN OUTCOME MEASURE(S): Telomere length and telomere aberrations were analyzed after telomere and centromere staining.RESULT(S): The mean telomere length of patients consulting for infertility was significantly less than that of healthy donors of similar age. Moreover, patients with infertility showed significantly more extreme telomere loss and telomere doublet formation than healthy controls. Telomere shortening and/or telomere aberrations were more pronounced in patients with structural chromosomal aberrations. Dicentric chromosomes were identified in 6/13 patients, with constitutional chromosomal aberrations leading to chromosomal instability that correlated with chromosomal end-to-end fusions.CONCLUSION(S): Our findings demonstrate the feasibility of analyzing telomere aberrations in addition to chromosomal aberrations, using cytogenetic slides. Telomere attrition and/or dysfunction represent the main common cytogenetic characteristic of patients with infertility, leading to potential implications for fertility assessment. Pending further studies, these techniques that correlate the outcome of assisted reproduction and telomere integrity status may represent a novel and useful diagnostic and/or prognostic tool for medical care in this field.

In the event of a radiological or nuclear accident, or when physical dosimetry is not available, the scoring of radiation-induced chromosomal aberrations in lymphocytes constitutes an essential tool for the estimation of the absorbed dose of the exposed individual and for effective triage. Cytogenetic biodosimetry employs different cytogenetic assays including the scoring of dicentrics, micronuclei, and translocations as well as analyses of induced premature chromosome condensation to define the frequency of chromosome aberrations. However, inherent challenges using these techniques include the considerable time span from sampling to result, the sensitivity and specificity of the various techniques, and the requirement of highly skilled personnel. Thus, techniques that obviate these challenges are needed. The introduction of telomere and centromere (TC) staining have successfully met these challenges and, in addition, greatly improved the efficiency of cytogenetic biodosimetry through the development of automated approaches, thus reducing the need for specialized personnel. Here, we review the role of the various cytogenetic dosimeters and their recent improvements in the management of populations exposed to genotoxic agents such as ionizing radiation. Finally, we discuss the emerging potentials to exploit these techniques in a wider spectrum of medical and biological applications, e.g., in cancer biology to identify prognostic biomarkers for the optimal triage and treatment of patients.

International audience; The recent development of Internet of Things (IoT) has democratized activity monitoring. Even if the data collected can be useful for healthcare, sharing this sensitive information exposes users to privacy threats and re-identification. This paper presents two approaches to anonymize the motion sensor data. The first is an extension of an earlier work based on filtering in the time-frequency plane and convolutional neural network; and the second is based on handcrafted features extracted from the zeros distribution of the time-frequency representation. The two approaches are evaluated on a public dataset to assess the accuracy of activity recognition and user re-identification. With the first approach we obtained an accuracy rate in activity recognition of 73% while limiting the identity recognition to an accuracy rate of 30% which corresponds to an activity identity ratio of 2.4. With the second approach we succeeded in improving the activity and identity ratio to 2.67 by attaining an accuracy rate in activity recognition of 80% while maintaining the re-identification rate at 30%.

Pandemics and lockdowns may be associated with unpremeditated consequences, such as bodyweight changes, isolation, as well as sedentarity. Reports have been published on malnutrition among patients suffering from COVID-19. This study aimed to highlight the short-term effects of the lockdown on the nutritional health of elderly people living at home and benefiting from home care services, yet without any COVID-19 pathology. In 50 subjects displaying weight, body mass index, and MNA score stability two months earlier, we observed significant alterations in these parameters following the lockdown. Thus, malnutrition rose from 28–34% to 58%. Furthermore, trigger factors for malnutrition changed, with social isolation accounting for 64% of the confinement’s deleterious effects among the elderly. In conclusion, despite the elderly being not directly affected by SARS-CoV2, the nutritional status of elderly subjects living at home with no or only mild autonomy loss was greatly and rapidly affected by the lockdown. Moreover, the main trigger factors for malnutrition were essentially related to social isolation and depressive syndromes. Knowing the impact of confinement on the elderly’s health, these results may help further modulate ongoing public health interventions in case of future lockdowns.

Light-field and plenoptic cameras are widely available today. Compared with monocular cameras, these cameras capture not only the intensity but also the direction of the light rays. Due to this specificity, light-field cameras allow for image refocusing and depth estimation using a single image. However, most of the existing depth estimation methods using light-field cameras require a prior complex calibration phase and raw data preprocessing before the desired algorithm is applied. We propose a homography-based method with plenoptic camera parameters calibration and optimization, dedicated to our homography-based micro-images matching algorithm. The proposed method works on debayerred raw images with vignetting correction. The proposed approach directly links the disparity estimation in the 2D image plane to the depth estimation in the 3D object plane, allowing for direct extraction of the real depth without any intermediate virtual depth estimation phase. Also, calibration parameters used in the depth estimation algorithm are directly estimated, and hence no prior complex calibration is needed. Results are illustrated by performing depth estimation with a focused light-field camera over a large distance range up to 4 m.

International audience; Purpose: Biological dosimetry, based on the relationship between the absorbed dose after exposure to ionizing radiation and the frequency of scored aberrations, has been and continues to be an important tool for estimating the dose after exposure. Dicentric chromosomes are considered to be the most specific and sensitive aberration related to radiation exposure. Here, we established the dose-response curve following in vitro irradiation of circulating lymphocytes from healthy donors from three African countries after scoring unstable chromosomal aberrations.Materials and methods: Blood samples from 16 African donors were exposed to various doses (0 to 4 Gy) using an X-RAD320 x-ray system with a maximum photon energy of 250 kV at a dose rate of 0.1 Gy min−1. Blood lymphocytes were cultured for 48 h, and chromosomal aberrations were scored during the first mitosis by telomere and centromere staining. The distribution of dicentric chromosomes was determined. Results: No dicentric chromosomes were found after the analysis of 2,669 first-division metaphases before in vitro exposure. We established a linear-quadratic dose-response curve based on the frequency of dicentric and ring chromosomes and calculated double-strand breaks, taking into account all scored aberrations.Conclusion: The generation of a specific dose-response curve for African donors will allow the practice of precise biological dosimetry in these countries. This work is the first step towards realizing an African biodosimetry network and the establishment of a biological dosimetry laboratory, which could play a major role in the application of radioprotection norms.

Telomeres play a major role in maintaining genome stability and integrity. Putative involvement of telomere dysfunction in the formation of various types of chromosomal aberrations is an area of active research. Here, we report a case of a six-month-old boy with a chromosomal gain encompassing the 11q22.3q25 region identified by SNP array analysis. The size of the duplication is 26.7 Mb and contains 170 genes (OMIM). The duplication results in partial trisomy of the region in question with clinical consequences, including bilateral renal dysplasia, delayed development, and a heart defect. Moreover, the karyotype determined by R-banding and chromosome painting as well as by hybridization with specific sub-telomere probes revealed the presence of an unbalanced t(9;11)(p24;q22.3) translocation with a unique breakpoint involving the sub-telomere region of the short arm of chromosome 9. The karyotypes of the parents were normal. Telomere integrity in circulating lymphocytes from the child and from his parents was assessed using an automated high-throughput method based on fluorescence in situ hybridization (FISH) with telomere- and centromere-specific PNA probes followed by M-FISH multicolor karyotyping. Very short telomeres, as well as an increased frequency of telomere loss and formation of telomere doublets, were detected in the child’s cells. Interestingly, similar telomere profiles were found in the circulating lymphocytes of the father. Moreover, an assessment of clonal telomere aberrations identified chromosomes 9 and 11 with particularly high frequencies of such aberrations. These findings strongly suggest that telomere dysfunction plays a central role in the formation of this specific unbalanced chromosome rearrangement via chromosome end-to-end fusion and breakage–fusion–bridge cycles.

Light-Field (LF) cameras allow the extraction not only of the intensity of light but also of the direction of light rays in the scene, hence it records much more information of the scene than a conventional camera. In this paper, we present a novel method to detect key-points in raw LF images by applying key-points detectors on Pseudo-Focused images (PFIs). The main advantage of this method is that we don’t need to use complex key-points detectors dedicated to light-field images. We illustrate the method in two use cases: the extraction of corners in a checkerboard and the key-points matching in two view raw light-field images. These key-points can be used for different applications e.g. calibration, depth estimation or visual odometry. Our experiments showed that our method preserves the accuracy of detection by re-projecting the pixels in the original raw images.

Background: Despite significant improvements in COVID-19 therapy, many patients still present with persistent symptoms and quality-of-life alterations. The aim of this study was to simultaneously investigate the long-term evolution of autonomy, malnutrition and long-lasting symptoms in people infected with COVID-19 and hospitalized in the ICU. Method: Patients’ clinical characteristics; extent of their loss of autonomy based on “Autonomie Gérontologie Groupes Iso-Ressources” (AG-GIR) classification; nutritional status while following the French and Global Leadership Initiative on Malnutrition (GLIM) recommendations; and symptom evolutions before infection, during hospitalization and rehabilitation, and up to 6 months after returning home were determined in thirty-seven patients. Results: Prior to a COVID-19 infection, all patients were autonomous, but upon admission to the rehabilitation center (CRM), 39% of them became highly dependent. After discharge from the center and 6 months after returning home, only 6 and 3%, respectively, still required considerable assistance. Of these thirty-seven patients, 11% were moderately malnourished and 81% presented with severe malnutrition, with a significant correlation being observed between malnutrition and autonomy (p < 0.05). Except for fatigue, which persisted in 70% of the patients 6 months after discharge from rehabilitation, all other symptoms decreased significantly. Conclusions: This study shows a striking decrease in autonomy associated with malnutrition after hospitalization for a COVID-19 infection and a clear beneficial effect from personalized rehabilitation. However, although almost all patients regained autonomy 6 months after returning home, they often still suffer from fatigue. Patient compliance with their nutritional recommendations deserves further improvement, preferably through personalized and persistent follow-up with the patient.

Dicentric chromosomes are a relevant marker of chromosomal instability. Their appearance is associated with telomere dysfunction, leading to cancer progression and a poor clinical outcome. Here, we present Telomere and Centromere staining followed by M-FISH (TC+M-FISH) for improved detection of telomere dysfunction and the identification of dicentric chromosomes in cancer patients and various genetic syndromes. Significant telomere length shortening and significantly higher frequencies of telomere loss and deletion were found in the peripheral lymphocytes of patients with cancer and genetic syndromes relative to similar age-matched healthy donors. We assessed our technique against conventional cytogenetics for the detection of dicentric chromosomes by subjecting metaphase preparations to both approaches. We identified dicentric chromosomes in 28/50 cancer patients and 21/44 genetic syndrome patients using our approach, but only 7/50 and 12/44, respectively, using standard cytogenetics. We ascribe this discrepancy to the identification of the unique configuration of dicentric chromosomes. We observed significantly higher frequencies of telomere loss and deletion in patients with dicentric chromosomes (p &lt, 10&minus, 4). TC+M-FISH analysis is superior to classical cytogenetics for the detection of chromosomal instability. Our approach is a relatively simple but useful tool for documenting telomere dysfunction and chromosomal instability with the potential to become a standard additional diagnostic tool in medical genetics and the clinic.

SARS-CoV-2 is generating a crisis in health, food, and nutrition. This is particularly the case in patients with chronic illnesses, such as diabetic patients, the elderly and patients living in social and financial precariousness. So despite the many remaining uncertainties that are profoundly changing people and habits, there is undeniably good reason not only to detect, correct, educate, and implement corrective measures, but also to carry out research and assessment.

A link between telomere shortening and oxidative stress was found in aging people and patients with cancer or inflammatory diseases. Extracts of Astragalus spp. are known to stimulate telomerase activity, thereby compensating telomere shortening. We characterized a multi-component hydroethanolic root extract (HRE) of Astragalus mongholicus Bunge and assessed its effects on telomeres compared to those of danazol. Astragalosides I to IV, flavonoids, amino acids and sugars were detected in the HRE. Samples of peripheral blood lymphocytes with short telomeres from 18 healthy donors (mean age 63.5 years, range 3286 years) were exposed to a single dose of 1 &micro, g/mL HRE or danazol for three days. Telomere length and telomerase expression were then measured. Significant elongation of telomeres associated to a less toxicity was observed in lymphocytes from 13/18 donors following HRE treatment (0.54 kb (0.15&ndash, 2.06 kb)) and in those from 9/18 donors after danazol treatment (0.95 kb (0.06&ndash, 2.06 kb)). The rate of cells with short telomeres (&lt, 3 kb) decreased in lymphocytes from all donors after exposure to either HRE or danazol, telomere elongation being telomerase-dependent. These findings suggest that the HRE could be used for the management of age-related diseases.

A link between telomere shortening and oxidative stress was found in aging people and patients with cancer or inflammatory diseases. Extracts of Astragalus spp. are known to stimulate telomerase activity, thereby compensating telomere shortening. We characterized a multi-component hydroethanolic root extract (HRE) of Astragalus mongholicus Bunge and assessed its effects on telomeres compared to those of danazol. Astragalosides I to IV, flavonoids, amino acids and sugars were detected in the HRE. Samples of peripheral blood lymphocytes with short telomeres from 18 healthy donors (mean age 63.5 years; range 32–86 years) were exposed to a single dose of 1 µg/mL HRE or danazol for three days. Telomere length and telomerase expression were then measured. Significant elongation of telomeres associated to a less toxicity was observed in lymphocytes from 13/18 donors following HRE treatment (0.54 kb (0.15–2.06 kb)) and in those from 9/18 donors after danazol treatment (0.95 kb (0.06–2.06 kb)). The rate of cells with short telomeres (

Background The cytokinesis-block micronucleus (CBMN) assay is an internationally recognized method for measuring DNA damage after exposure to genotoxic agents, as well as a biomarker for DNA repair and chromosomal instability. The high baseline level of micronuclei (MN) in the healthy population has limited the sensitivity and application of the CBMN assay for the follow-up of exposed populations. We reevaluated the sensitivity of the CBNM assay using semi-automated MN scoring following telomere and centromere (TC) staining after in vitro exposure to genotoxic agents (mitomycin or radiation) or aneugenic agents (vinblastine). Materials and methods Blood samples from 12 healthy donors were exposed to 137Cs at seven doses from 0.1–4 Gy and cultured for 72 h. Cytochalasin B was added at 46 h of culture. The exposure of chemical agents (mitomycin or vinblastine) was performed after 48 h of culture for 3 h. Cytochalasin B was added after treatment and slides were prepared 24 h after. MN was semi-automatically scored following TC staining. Nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs) were assessed in a human cell line after TC staining. Results The introduction TC staining to the scoring of MN not only renders MN scoring more efficient and robust, but also permits discrimination between exposure to clastogenic (MN with only telomere signals) and aneugenic agents (MN with both TC signals). The resulting improvement of MN detection led to an increase in the sensitivity of the CBMN assay following low-dose radiation exposure (0.3 versus 0.1 Gy). Hyperradiosensitivity phenomenon was observed after low dose exposure. A dose-response curve was obtained for up to 4 Gy. In addition, TC staining permits assessment of the nature of NPBs and NBUDs as biomarkers for genotoxicity and chromosomal instability. Conclusion These approaches can be potentially used to follow-up populations exposed to genotoxic agents and assess cancer risk.

The authors wish to make the following corrections to this paper [...]

The ability of parabens to promote the appearance of multiple cancer hallmarks in breast epithelium cells provides grounds for regulatory review of the implication of the presence of parabens in human breast tissue. It is well documented that telomere dysfunction plays a significant role in the initiation of genomic instability during carcinogenesis in human breast cancer. In the present study, we evaluated the genotoxic effect of ethyl 4-hydroxybenzoate (ethyl-paraben), with and without metabolic activation (S9), in studies following OECD guidelines. We observed a significant increase in genotoxic damage using the Mouse Lymphoma Assay and in vitro micronucleus (MN) tests in the L5178Y cell line in the presence of S9 only after a short exposure. A high frequency of MN was observed in the TK6 cells after a short exposure (3 h) in the presence of S9 and a long exposure (26 h) without S9. We found significant increases in the MN frequency and induced chromosomal aberrations in the lymphocytes of only one donor after ethyl-paraben exposure in the presence of S9 after a short exposure. Cytogenetic characterization of the paraben-treated cells demonstrated telomere shortening associated with telomere loss and telomere deletions in L5178Y and TK6 cells and lymphocytes of the paraben sensitive-donor. In a control cohort of 68 human lymphocytes, telomere length and telomere aberrations were age-dependent and showed high inter-individual variation. This study is the first to link telomere shortening and the genotoxic effect of ethyl paraben in the presence of S9 and raises the possibility that telomere shortening may be a proxy for underlying inter-individual sensitivity to ethyl-paraben. Copyright © 2016 John Wiley & Sons, Ltd.

Resume Introduction La telemedecine est susceptible d’apporter une aide a la prise en charge des patients souffrant d’insuffisance cardiaque et d’eviter des urgences vitales. Dans ce cadre, nous avons experimente dans une unite de medecine interne une e-plateforme E-care dediee a la detection automatisee et intelligente des situations a risque de decompensation cardiaque. Methodes La plateforme E-care repose sur des capteurs medicaux (pression arterielle, frequence cardiaque, O2, poids), communicants (Bluetooth), permettant de remonter, en temps reel, a un systeme intelligent des informations physiologiques et sur une analyse de l’ontologie medicale, ce qui aboutit in fine a la generation d’alertes. Apres une phase de mise au point (proof of concept), la plateforme E-care a ete deployee et testee par les professionnels de sante et les patients dans une unite de medecine interne de 20 lits, ouverte sur les urgences aux hopitaux universitaires de Strasbourg. Resultats Cent quatre-vingts patients ont ete inclus et 1500 mesures ont ete realisees. Le profil de patients inclus dans cette experimentation etait un patient âge, poly-pathologique dans plus de 90 % des cas, avec une perte d’autonomie dans 25 %. Les professionnels de sante utilisaient la plateforme E-care au quotidien a leur grande satisfaction. Cette experimentation permettait de valider les choix technologiques, de consolider le systeme et de tester la robustesse de la plateforme E-care. La collecte en continu nous a permis d’avoir le nombre critique de patients pour analyser plus finement la pertinence des alertes en rapport avec des decompensations cardiaques. Une analyse preliminaire montrait la pertinence des alertes generees. Conclusion Les resultats preliminaires suite au deploiement de la plateforme E-care en milieu hospitalier semblent montrer la pertinence des choix technologiques, des outils et des solutions developpees et adoptees. Ce systeme de telemedecine permet de facon automatique, non intrusive, de generer des alertes en rapport avec la detection de situations a risque de decompensation cardiaque, etant in fine susceptible de prevenir le recours a l’hospitalisation. Un deploiement a domicile est actuellement en cours.

International audience; Classifying biofuel feedstock crops on the basis of their inorganic constituents is needed to evaluate the critical role these components play during combustion and, later, when the ashes are used as fertilizer or as secondary raw material. Fluorescent and white-light microscopy, in combination with environmental scanning electron microscopy and energy-dispersive X-ray spectroscopy, was used to characterize phytoliths in Miscanthus sinensis, its dry extracts, and in the cyclone ash produced by the semi-industrial (400 kW) burning of this energy crop. The results show that the phytoliths are made of opal, SiO2•n(H2O). X-ray powder diffraction data document that the SiO2-rich (31.7 wt%) cyclone ash consists of nearly 83 wt% amorphous material, which reflects the abundance of phytoliths in the studied materials. The cyclone ash also contains high amounts of some heavy metals, which suggests that Miscanthus sinensis is suitable for bioremediation of contaminated soils.

Inhalation of particulate matter (PM) from residential biomass combustion is epidemiologically associated with cardiovascular and pulmonary diseases. This study investigates PM0.4-1 emissions from combustion of commercial Miscanthus straw (MS), softwood chips (SWC) and beech wood chips (BWC) in a domestic-scale boiler (40 kW). The PM0.4-1 emitted during combustion of the MS, SWC and BWC were characterized by ICP-MS/OES, XRD, SEM, TEM, and DLS. Cytotoxicity and genotoxicity in human alveolar epithelial A549 and human bronchial epithelial BEAS-2B cells were assessed by the WST-1 assay and the DNA-Alkaline Unwinding Assay (DAUA). PM0.4-1 uptake/translocation in cells was investigated with a new method developed using a confocal reflection microscope. SWC and BWC had a inherently higher residual water content than MS. The PM0.4-1 emitted during combustion of SWC and BWC exhibited higher levels of Polycyclic Aromatic Hydrocarbons (PAHs), a greater variety of mineral species and a higher heavy metal content than PM0.4-1 from MS combustion. Exposure to PM0.4-1 from combustion of SWC and BWC induced cytotoxic and genotoxic effects in human alveolar and bronchial cells, whereby the strongest effect was observed for BWC and was comparable to that caused by diesel PM (SRM 2 975), In contrast, PM0.4-1 from MS combustion did not induce cellular responses in the studied lung cells. A high PAH content in PM emissions seems to be a reliable chemical marker of both combustion efficiency and particle toxicity. Residual biomass water content strongly affects particulate emissions and their toxic potential. Therefore, to minimize the harmful effects of fine PM on health, improvement of combustion efficiency (aiming to reduce the presence of incomplete combustion products bound to PM) and application of fly ash capture technology, as well as use of novel biomass fuels like Miscanthus straw is recommended.

To identify the cells responsible for the initiation and maintenance of Hodgkin lymphoma (HL) cells, we have characterized a subpopulation of HL cells grown in vitro and in vivo with the aim of establishing a reliable and robust animal model for HL. To validate our model, we challenged the tumor cells in vivo by injecting the alkylating histone-deacetylase inhibitor, EDO-S101, a salvage regimen for HL patients, into xenografted mice. Methodology: Blood lymphocytes from 50 HL patients and seven HL cell lines were used. Immunohistochemistry, flow cytometry, and cytogenetics analyses were performed. The in vitro and in vivo effects of EDO-S101 were assessed. Results: We have successfully determined conditions for in vitro amplification and characterization of the HL L428-c subline, containing a higher proportion of CD30&minus, /CD15&minus, cells than the parental L428 cell line. This subline displayed excellent clonogenic potential and reliable reproducibility upon xenografting into immunodeficient NOD-SCID-gamma (&minus, /&minus, )(NSG) mice. Using cell sorting, we demonstrate that CD30&minus, subpopulations can gain the phenotype of the L428-c cell line in vitro. Moreover, the human cells recovered from the seventh week after injection of L428-c cells into NSG mice were small cells characterized by a high frequency of CD30&minus, cells. Cytogenetic analysis demonstrated that they were diploid and showed high telomere instability and telomerase activity. Accordingly, chromosomal instability emerged, as shown by the formation of dicentric chromosomes, ring chromosomes, and breakage/fusion/bridge cycles. Similarly, high telomerase activity and telomere instability were detected in circulating lymphocytes from HL patients. The beneficial effect of the histone-deacetylase inhibitor EDO-S101 as an anti-tumor drug validated our animal model. Conclusion: Our HL animal model requires only 103 cells and is characterized by a high survival/toxicity ratio and high reproducibility. Moreover, the cells that engraft in mice are characterized by a high frequency of small CD30&minus, cells exhibiting high telomerase activity and telomere dysfunction.

Background: We analyzed telomere maintenance mechanisms (TMMs) in lymph node samples from HL patients treated with standard therapy. The TMMs correlated with clinical outcomes of patients. Materials and Methods: Lymph node biopsies obtained from 38 HL patients and 24 patients with lymphadenitis were included in this study. Seven HL cell lines were used as in vitro models. Telomerase activity (TA) was assessed by TRAP assay and verified through hTERT immunofluorescence expression, alternative telomere lengthening (ALT) was also assessed, along with EBV status. Results: Both TA and ALT mechanisms were present in HL lymph nodes. Our findings were reproduced in HL cell lines. The highest levels of TA were expressed in CD30&minus, /CD15&minus, cells. Small cells were identified with ALT and TA. Hodgkin and Reed Sternberg cells contained high levels of PML bodies, but had very low hTERT expression. There was a significant correlation between overall survival (p &lt, 10&minus, 3), event-free survival (p &lt, 4), and freedom from progression (p &lt, 3) and the presence of an ALT profile in lymph nodes of EBV+ patients. Conclusion: The presence of both types of TMMs in HL lymph nodes and in HL cell lines has not previously been reported. TMMs correlate with the treatment outcome of EBV+ HL patients.

International audience; Background: Microsatellite and chromosomal instability have been investigated in Hodgkin lymphoma (HL). Materials and Methods: We studied seven HL cell lines (five Nodular Sclerosis (NS) and two Mixed Cellularity (MC)) and patient peripheral blood lymphocytes (100 NS-HL and 23 MC-HL). Microsatellite instability (MSI) was assessed by PCR. Chromosomal instability and telomere dysfunction were investigated by FISH. DNA repair mechanisms were studied by transcriptomic and molecular approaches. Results: In the cell lines, we observed high MSI in L428 (4/5), KMH2, and HDLM2 (3/5), low MSI in L540, L591, and SUP-HD1, and none in L1236. NS-HL cell lines showed telomere shortening, associated with alterations of nuclear shape. Small cells were characterized by telomere loss and deletion, leading to chromosomal fusion, large nucleoplasmic bridges, and breakage/fusion/bridge (B/F/B) cycles, leading to chromosomal instability. The MC-HL cell lines showed substantial heterogeneity of telomere length. Intrachromosmal double strand breaks induced dicentric chromosome formation, high levels of micronucleus formation, and small nucleoplasmic bridges. B/F/B cycles induced complex chromosomal rearrangements. We observed a similar pattern in circulating lymphocytes of NS-HL and MC-HL patients. Transcriptome analysis confirmed the differences in the DNA repair pathways between the NS and MC cell lines. In addition, the NS-HL cell lines were radiosensitive and the MC-cell lines resistant to apoptosis after radiation exposure. Conclusions: In mononuclear NS-HL cells, loss of telomere integrity may present the first step in the ongoing process of chromosomal instability. Here, we identified, MSI as an additional mechanism for genomic instability in HL.