The development of a hiPSC-based platform to identify tissue-dependencies of IDH1 R132H.

The application of patient-derived (PD) in vitro tumor models represents the classical strategy for clinical translational oncology research. Using these cellular heterogeneous cultures for the isolation of cancer stem cells (CSCs), suggested to be the main driver for disease malignancy, relies on the use of surrogate biomarkers or is based on CSC-enriching culture conditions. However, the ability of those strategies to exclusively and efficiently enrich for CSC pool has been questioned. Here we present an alternative in vitro CSC model based on the oncogenic transformation of single clone-derived human induced pluripotent stem cells (hiPSC). Hotspot mutations in the DNA encoding for the R132 codon of the enzyme isocitrate dehydrogenase 1 (IDH1) and codon R175 of p53 are commonly occurring molecular features of different tumors and were selected for our transformation strategy. By choosing p53 mutant glial tumors as our model disease, we show that in vitro therapy discovery tests on IDH1-engineered synthetic CSCs (sCSCs) can identify kinases-targeting chemotherapeutics that preferentially target tumor cells expressing corresponding genetic alteration. In contrast, neural stem cells (NSCs) derived from the IDH1R132H overexpressing hiPSCs increase their resistance to the tested interventions indicating glial-to-neural tissue-dependent differences of IDH1R132H. Taken together, we provide proof for the potential of our sCSC technology as a potent addition to biomarker-driven drug development projects or studies on tumor therapy resistance. Moreover, follow-up projects such as comparing in vitro drug sensitivity profiles of hiPSC-derived tissue progenitors of different lineages, might help to understand a variety of tissue-related functions of IDH1 mutations.

Contribution of the Synkinesis Assessment Questionnaire and the Sunnybrook Facial Grading System to the evaluation of synkinesis after peripheral facial palsy: A STROBE observational study.

OBJECTIVES: The main aim of the study was to determine whether the perception of synkinesis by patients with peripheral facial palsy (PFP) matched their clinician's severity assessment. Secondary objectives comprised: (1) to determine whether objective measurement of synkinesis matched the patient's perception; and (2) is to identify factors influencing patients' perceptions. METHODS: This retrospective study took place from January to May 2020. Forty patients (8 per PFP grade, I-V/VI; 20 women, 20 men) filled out the Synkinesis Assessment Questionnaire (SAQ) and were assessed on the Sunnybrook Facial Grading System (SFGS). Photographs were analyzed on MEEI-Facegram software. RESULTS: Perceived synkinesis (total SAQ) matched objective grades (SFGS) (Z=2.89; P=0.004), especially for smiling (Z=3.84; P<0.001) and lip protrusion (Z=3.79; P<0.001). Synkinesis on lip protrusion was a more sensitive indicator of perceived synkinesis than synkinesis on smiling (Z=2.96; P=0.003). Duration (ρ=0.5137; P<0.001) and grade of PFP (Chi2=13.82; P=0.008) heightened the perception of synkinesis. CONCLUSION: Patient-reported outcome measures (PROMs) such as the SAQ are relevant for clinical evaluation.

Longitudinal stability of molecular alterations and drug response profiles in tumor spheroid cell lines enables reproducible analyses.

The utility of patient-derived tumor cell lines as experimental models for glioblastoma has been challenged by limited representation of the in vivo tumor biology and low clinical translatability. Here, we report on longitudinal epigenetic and transcriptional profiling of seven glioblastoma spheroid cell line models cultured over an extended period. Molecular profiles were associated with drug response data obtained for 231 clinically used drugs. We show that the glioblastoma spheroid models remained molecularly stable and displayed reproducible drug responses over prolonged culture times of 30 in vitro passages. Integration of gene expression and drug response data identified predictive gene signatures linked to sensitivity to specific drugs, indicating the potential of gene expression-based prediction of glioblastoma therapy response. Our data thus empowers glioblastoma spheroid disease modeling as a useful preclinical assay that may uncover novel therapeutic vulnerabilities and associated molecular alterations.

[Sequelae in bell's palsy: Prognostic factors for recovery]. / Facteurs prédictifs des séquelles de paralysie faciale idiopathique : suivi longitudinal.

OBJECTIVES: Facial palsy can be assessed using objective and subjective tools. The main purpose of this work was to use these tools to determine at 12 months the percentage of patients with sequelae and to specify the type of sequelae. MATERIAL AND METHODS: Twenty-three patients with facial palsy were followed in this prospective and longitudinal study. They have been evaluated every 3 months during a year with the House and Brackmann grading scale and the Sunnybrook Facial Grading System. At 12 months, group A was composed of patients with complete recovery and group B, patients with sequelae. RESULTS: At 3 months, in patients of group B, the House-Brackmann grading scale (P=0.0134), the Sunnybrook Facial Grading System global score (P=0.0283) and dynamic score (P=0.0148) were lower than group A. Moreover, the movement "brow lift" (P=0.0181) seems to be relevant to predict follow-up. Synkinesis on "brow lift" (P=0.0270) and the treatment delay (P=0.0384) increased for group B. Sex, age, paralyzed side and recurrence of facial palsy had no influence. CONCLUSION: Objective and subjective tools determine thresholds and predictive scores of recovery with sequelae at 1 year. Nevertheless, it is relevant to assess clinically specific facial movements, such as "brow lift", to specify a recovery potential and to predict sequelae a year after the onset of facial palsy. As the treatment delay influences recovery, drug treatment should be recommended as early as possible.

Fine-scale genetic structure in a high dispersal capacity raptor, the Montagu's harrier (Circus pygargus), revealed by a set of novel microsatellite loci.

The Montagu's harrier (Circus pygargus) is a semi-colonial raptor species widely but patchily distributed across the Palearctic region with recorded cases of philopatry and presence of extra-pair copulation. In order to assess Montagu's harrier spatial genetic structure and contemporary gene flow, we developed 16 new microsatellite markers using 454 pyrosequencing. Genotypes of 117 chicks sampled in a 200 × 300 km farmland area in Central Western France were analyzed to characterize genetic polymorphism at each locus and regional and fine-scale genetic structure. Fourteen markers were found polymorphic, with a number of alleles ranging from 3 to 11. The expected and observed heterozygosities ranged from 0.36 to 0.856 and from 0.35 to 0.868, respectively. A single genetic unit was found at the regional scale with higher genetic similarity observed at a small spatial scale (up to 10 km). Our results are consistent with overall large-scale juvenile and adult dispersal together with small-scale male philopatry. Cross-species amplification of this set of microsatellites makers has been successful in two closely related harrier species: the marsh harrier (Circus aeruginosus) and the Hen harrier (Circus cyaneus) for which 14 and 12 markers were polymorphic, respectively. These new microsatellite markers could be used to study the population genetic structure, contemporary gene flow and parentage analyses in these three species and to conduct microsatellite-based demographic inferences on the Montagu's harrier.

CBF1 is clinically prognostic and serves as a target to block cellular invasion and chemoresistance of EMT-like glioblastoma cells.

BACKGROUND: Glioblastoma is the most common and most lethal primary brain cancer. CBF1 (also known as Recombination signal Binding Protein for immunoglobulin kappa J, RBPJ) is the cardinal transcriptional regulator of the Notch signalling network and has been shown to promote cancer stem-like cells (CSCs) in glioblastoma. Recent studies suggest that some of the malignant properties of CSCs are mediated through the activation of pro-invasive programme of epithelial-to-mesenchymal transition (EMT). Little is known whether CBF1 is involved in the EMT-like phenotype of glioma cells. METHODS: In a collection of GBM neurosphere lines, we genetically inhibited CBF1 and investigated the consequences on EMT-related properties, including in vitro invasiveness by Boyden chambers assay, chemoresistance using a clinical drug library screen and glycolytic metabolism assessing live-cell extracellular acidification rate. We also compared CBF1 expression in cells exposed to low and high oxygen tension. In silico analysis in large-scale Western and Eastern patient cohorts investigated the clinical prognostic value of CBF1 expression in low- and high-grade glioma as well as medulloblastoma. RESULTS: Mean CBF1 expression is significantly increased in isocitrate dehydrogenase 1 (IDH1) R132H mutant glioblastoma and serves as prognostic marker for prolonged overall survival in brain tumours, particularly after therapy with temozolomide. Hypoxic regions of glioblastoma have higher CBF1 activation and exposure to low oxygen can induce its expression in glioma cells in vitro. CBF1 inhibition blocks EMT activators such as zinc finger E-box-binding homeobox 1 (ZEB1) and significantly reduces cellular invasion and resistance to clinically approved anticancer drugs. Moreover, we indicate that CBF1 inhibition can impede cellular glycolysis. CONCLUSIONS: Mean CBF1 activation in bulk tumour samples serves as a clinical predictive biomarker in brain cancers but its intratumoral and intertumoral expression is highly heterogeneous. Microenvironmental changes such as hypoxia can stimulate the activation of CBF1 in glioblastoma. CBF1 blockade can suppress glioblastoma invasion in vitro in particular in cells undergone EMT such as those found in the hypoxic niche. Targeting CBF1 can be an effective anti-EMT therapy to impede invasive properties and chemosensitivity in those cells.

Advances in methods for estimating stopover duration for migratory species using capture-recapture data.

Many species are migratory, resulting in a life cycle divided into periodic stages occurring in different habitats occupied for a limited amount of time. Estimating the time spent in each habitat is crucial to understanding how individuals modulate their activities and thus to evolutionary ecology and conservation biology. Several methods, including some recent promising advances, can be used to estimate stopover duration as well as arrival and departure probabilities at sites where individuals are monitored using capture-recapture sampling. Our objectives in this study were to (1) describe the available models to estimate stopover duration, (2) illustrate with an original data set what kinds of questions can be addressed using the most recent methods, and (3) to provide in a detailed appendix a practical guide for implementing these methods in E-SURGE software. To illustrate the potential of these models for testing biological hypotheses, we used a capture-recapture data set on marbled newts (Triturus marmoratus). We used time-dependent and time-elapsed-since-arrival effects (using both Markovian and semi-Markov processes for the latter) to model stopover duration and the probability of arriving in and departing from a breeding pond for this species and compared the relative performance of the resulting models. Our findings showed a strong sex effect on stopover duration: females stayed on average 5.63 weeks in a breeding pond whereas males stayed only 3.03 weeks. In both sexes, the retention probability was mainly influenced by the time already spent there. Consequently, individuals of the same sex stayed a similar amount of time in a pond, although they did not arrive simultaneously but successively. The selected data set demonstrated the flexibility of these methods and their potential relevance for applications in evolutionary ecology and conservation.

Recurrence of drug-induced reactions in DRESS patients.

BACKGROUND: Drug reaction with eosinophilia and systemic symptoms (DRESS) may relapse following introduction of drugs structurally unrelated to the initial culprit drug. OBJECTIVE: To assess the frequency and characteristics of recurrent drug eruptions in patients with history of DRESS. METHODS: Patients who had developed adverse cutaneous reaction after DRESS occurrence were recruited from the regional database of Upper Normandy in France. Rate of recurrences were compared with patients with Toxic Epidermal Necrolysis (TEN) and Stevens-Johnson syndrome (SJS) patients during the same time frame. RESULTS: Of the 60 cases of DRESS collected, 15 (25%) with recurrences were retained for analysis. Seven patients had a single recurrence, whereas eight patients had several relapses. In the patients with pre-existing DRESS, recurrences were incomplete, corresponding to cutaneous rash in 13 cases and associated with eosinophilia in seven cases. Internal organ involvement was observed in two cases. In contrast, a single recurrence was found out of 61 patients with TEN/SJS. CONCLUSION: Incomplete recurrences with structurally unrelated culprit drugs are a frequent phenomenon in DRESS patients.

Increased prevalence of psoriasis in patients with coronary artery disease: results from a case-control study.

BACKGROUND: The incidence of myocardial events has been reported to be increased in patients with psoriasis. OBJECTIVES: To investigate whether psoriasis is an independent risk factor for coronary artery disease (CAD). METHODS: We compared the prevalence of psoriasis between case patients with a diagnosis of CAD based on coronary angiography findings and control patients with no CAD referred to the emergency surgery department for an acute noncardiovascular condition. Case and control patients were examined for the presence of psoriasis by two dermatologists. The prevalence of psoriasis was compared among patients with CAD according to CAD severity. Five-hundred cases and 500 age- and sex-matched controls were included. RESULTS: Using matched univariate analysis, the prevalence of psoriasis was about twofold higher in CAD case patients than in control patients [8·0% vs. 3·4%, odds ratio (OR) 2·64; 95% confidence interval (CI) 1·42-4·88]. Using unconditional multivariate analysis, the association of psoriasis with CAD appeared to be borderline significant (OR 1·84; 95% CI 0·99-3·40). Psoriasis in patients with CAD was significantly associated with three-vessel involvement relative to one-or two-vessel involvement (13·1% vs. 6·1%; OR 3·07; 95% CI 1·50-6·25). CONCLUSIONS: The prevalence of psoriasis is twofold higher in patients with CAD than in control patients without CAD. It is associated with a more severe coronary artery involvement.

Optimizing the trade-off between spatial and genetic sampling efforts in patchy populations: towards a better assessment of functional connectivity using an individual-based sampling scheme.

Genetic data are increasingly used in landscape ecology for the indirect assessment of functional connectivity, that is, the permeability of landscape to movements of organisms. Among available tools, matrix correlation analyses (e.g. Mantel tests or mixed models) are commonly used to test for the relationship between pairwise genetic distances and movement costs incurred by dispersing individuals. When organisms are spatially clustered, a population-based sampling scheme (PSS) is usually performed, so that a large number of genotypes can be used to compute pairwise genetic distances on the basis of allelic frequencies. Because of financial constraints, this kind of sampling scheme implies a drastic reduction in the number of sampled aggregates, thereby reducing sampling coverage at the landscape level. We used matrix correlation analyses on simulated and empirical genetic data sets to investigate the efficiency of an individual-based sampling scheme (ISS) in detecting isolation-by-distance and isolation-by-barrier patterns. Provided that pseudo-replication issues are taken into account (e.g. through restricted permutations in Mantel tests), we showed that the use of interindividual measures of genotypic dissimilarity may efficiently replace interpopulation measures of genetic differentiation: the sampling of only three or four individuals per aggregate may be sufficient to efficiently detect specific genetic patterns in most situations. The ISS proved to be a promising methodological alternative to the more conventional PSS, offering much flexibility in the spatial design of sampling schemes and ensuring an optimal representativeness of landscape heterogeneity in data, with few aggregates left unsampled. Each strategy offering specific advantages, a combined use of both sampling schemes is discussed.

Risk factors for abscess formation in patients with superficial cellulitis (erysipelas) of the leg.

BACKGROUND: Superficial cellulitis of the leg (erysipelas) is a frequent skin infection. Abscess formation is the most frequent local complication. Determinants of abscess formation in patients with leg cellulitis have not yet been clearly established. OBJECTIVE: To assess the risk factors for abscess formation in patients with leg cellulitis. METHODS: The clinical, biological and bacteriological records of all patients referred to the dermatology department of a university hospital for superficial cellulitis of the leg during a 3-year period were retrospectively reviewed. Using univariate and multivariate analysis, patients' main characteristics at baseline were compared between the group of patients who developed abscess and the group who did not. RESULTS: A total of 164 patients (93 female, 71 male), mean age 65±18 years, were included. Abscess occurred in 13 cases (8%). The following general factors were positively associated with abscess formation: male sex, smoking, alcohol abuse and delayed introduction of antibiotic treatment. Based on multivariate analysis, only chronic alcohol abuse [odds ratio (OR) 4·3, 95% confidence interval (CI)1·08-20·57] and delayed antibiotic treatment initiation (OR 1·4, 95% CI 1·02-2·04) remained independently associated with abscess formation. CONCLUSIONS: Alcohol abuse and delayed initiation of antibiotic treatment are risk factors for abscess formation in patients with cellulitis of the leg. Patients with these predictors must be monitored carefully for abscess formation.

[Clomipramine hypersensitivity with predominantly pulmonary involvement]. / DRESS syndrome avec atteinte pulmonaire prédominante sous clomipramine.

Drug hypersensitivity (DRESS syndrome) is a rare disorder with diverse systemic and visceral manifestations. Pulmonary involvement is uncommon and is mainly characterized by eosinophilic infiltration. We report a case of DRESS syndrome induced by clomipramine with predominant pulmonary involvement.

Skin swabbing as a new efficient DNA sampling technique in amphibians, and 14 new microsatellite markers in the alpine newt (Ichthyosaura alpestris).

This study introduces a novel DNA sampling method in amphibians using skin swabs. We assessed the relevancy of skin swabs relevancy for genetic studies by amplifying a set of 17 microsatellite markers in the alpine newt Ichthyosaura alpestris, including 14 new polymorphic loci, and a set of 11 microsatellite markers in Hyla arborea, from DNA collected with buccal swabs (the standard swab method), dorsal skin swabs and ventral skin swabs. We tested for quality and quantity of collected DNA with each method by comparing electrophoresis migration patterns. The consistency between genotypes obtained from skin swabs and buccal swabs was assessed. Dorsal swabs performed better than ventral swabs in both species, possibly due to differences in skin structure. Skin swabbing proved to be a useful alternative to buccal swabbing for small or vulnerable animals: by drastically limiting handling, this method may improve the trade-off between the scientific value of collected data, individual welfare and species conservation. In addition, the 14 new polymorphic microsatellites for the alpine newt will increase the power of genetic studies in this species. In four populations from France (n=19-25), the number of alleles per locus varied from 2 to 16 and expected heterozygosities ranged from 0.04 to 0.91. Presence of null alleles was detected in two markers and two pairs displayed gametic disequilibrium. No locus appeared to be sex-linked.

Haptic Recognition of Emotions in Raised-Line Drawings by Congenitally Blind and Sighted Adults.

15 sighted and 15 congenitally blind adults were to classify raised-line pictures of emotional faces through haptics. Whereas accuracy did not vary significantly between the two groups, the blind adults were faster at the task. These results suggest that raised-line pictures of emotional faces are intelligible to blind adults.

Occupational exposure to ambient electromagnetic fields of technical operational personnel working for a mobile telephone operator.

In order to investigate the exposure of operational personnel to radiofrequency electromagnetic fields when working for a mobile telephone operator, exposimeters were used to make individual records on 23 Technical Operations personnel (mobile telephone maintenance staff) and also on 22 Other Workers. The exposure densities, to which each of the 45 subjects was subjected, were quantified using 229 exposure indicators. Cluster analysis techniques were applied to the data, in an attempt to show that they would re-emerge as belonging to one of the two groups, i.e. the Technical Operational Personnel group or the Other Workers group. This exploratory investigation has shown that the cluster analysis does not reveal a sufficiently reliable emergence of the two groups, even though certain exposure indicators were significantly different for the two groups. In addition, the use of a Learning Group method does not lead to the discovery of a predictive law that could identify the Technical Operational Personnel as a sub-group within the overall group.

GATA4 is a regulator of astrocyte cell proliferation and apoptosis in the human and murine central nervous system.

The GATA transcription factors consist of six family members, which bind to the consensus DNA-binding element, W-GATA-R, and are poorly characterized in the central nervous system (CNS). Using retroviral gene trapping on transgenic mouse glioma models, we identified GATA6 to be a novel tumor suppressor gene in glioblastoma multiforme. We now show GATA4, a family member of GATA6, to be expressed in the neurons and glia of normal murine and human embryonic and adult CNS. Silencing GATA4 in normal astrocytes did not alter their growth properties. In contrast, knockdown of Gata4 in p53 null non-transformed murine astrocytes induced transformation, with increased proliferation and resistance to chemotherapy or radiation-induced apoptosis. Furthermore, GATA4 expression was lost in a panel of human malignant astrocytoma cell lines. GATA4 overexpression in normal human and murine astrocytes resulted in a cell cycle block in G1 phase, with increased apoptosis. Mechanistically, GATA4 was a transcriptional inducer of the cyclin-dependent kinase inhibitor, p15INK4B, leading to the attenuation of cyclin D1. GATA4 expression was also induced by transforming growth factor-beta, leading to the inhibition of astrocyte proliferation. Collectively, we show that GATA4 is expressed in the embryonic and adult CNS and acts as a negative regulator of astrocyte proliferation and growth.

A double-blind, placebo-controlled, randomized, multicenter study to investigate CHInese Medicine Neuroaid Efficacy on Stroke recovery (CHIMES Study).

UNLABELLED: Rationale Traditional Chinese Medications(TCM) have been reported to have beneficial effects in stroke patients, but were not rigorously evaluated by GCP standards. Aim This study tests the hypothesis that Neuroaid, a TCM widely used in China post-stroke, is superior to placebo in reducing neurological deficit and improving functional outcome in patients with acute cerebral infarction of an intermediate severity. Design This is a multicenter, randomised, double-blind, placebo-controlled study of Neuroaid in ischemic stroke patients with National Institute of Health Stroke Scale(NIHSS) 6-14 treated within 48 h of stroke onset. Neuroaid or placebo is taken (4 capsules) 3 times daily for 3 months. Treatments are assigned using block randomization, stratified for centers, via a central web-randomization system. With a power of 90% and two-sided test of 5% type I error, a sample size is 874. Allowing for a drop-out rate of up to 20%, 1100 individuals should be enrolled in this study. Study Outcomes The primary efficacy endpoint is the modified Rankin Scale(mRS) grades at 3 months. Secondary efficacy endpoints are the NIHSS score at 3 months; difference of NIHSS scores between baseline and 10 days, and between baseline and 3 months; difference of NIHSS sub-scores between baseline and 10 days, and between baseline and 3 months; mRS at 10 days, 1 month, and 3 months; Barthel index at 3 months; Mini Mental State Examination at 10 days and 3 months. Safety outcomes include complete blood count, renal and liver panels, and electrocardiogram. STUDY REGISTRATION: ClinicalTrials.gov identifier: NCT00554723.

Origin and genetic diversity of Western European populations of the potato cyst nematode (Globodera pallida) inferred from mitochondrial sequences and microsatellite loci.

Native to South America, the potato cyst nematode Globodera pallida is one of the principal pests of Andean potato crops and is also an important global pest following its introduction to Europe, Africa, North America, Asia and Oceania. Building on earlier work showing a clear south to north phylogeographic pattern in Peruvian populations, we have been able to identify the origin of Western European populations with high accuracy. They are all derived from a single restricted area in the extreme south of Peru, located between the north shore of the Lake Titicaca and Cusco. Only four cytochrome b haplotypes are found in Western Europe, one of them being also found in some populations of this area of southern Peru. The allelic richness at seven microsatellite loci observed in the Western European populations, although only one-third of that observed in this part of southern Peru, is comparable to the allelic richness observed in the northern region of Peru. This result could be explained by the fact that most of the genetic variability observed at the scale of a field or even of a region is already observed at the scale of a single plant within a field. Thus, even introduction via a single infected potato plant could result in the relatively high genetic variability observed in Western Europe. This finding has important consequences for the control of this pest and the development of quarantine measures.

Effects of exposing chicken eggs to a cell phone in "call" position over the entire incubation period.

The aim of the present study was to assess the effects of exposing fertile chicken eggs to a cell phone repeatedly calling a ten-digit number at 3-min intervals over the entire period of incubation. A pre-experiment was performed first to adjust incubation conditions in an experimental chamber devoid of metallic content and without automatic turning until the overall performance of hatchability was reproducible in the absence of the cell phone. The experimental period consisted of a series of 4 incubations referred to as "replicates". For each replicate, one batch of 60 eggs was exposed to the immediate environment (