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In order to further develop the potential applications of lignin biomass, the research on lignin nanoparticles (LNPs) and their nanocomposites has attracted increasing attention. In this study, a facile and no chemical modification approach to prepare stable alkali lignin nanospheres is presented. The nanospheres around 85-125 nm were prepared through the π-π interactions between molecules in the self-assembly process. Lignin alkali was dissolved in ethylene glycol at different initial concentrations and subsequently ultrasound and dialysis treatment were conducted to prepare LNPs. The prepared LNPs had zeta potentials between -20 mV and -40 mV, and they were electrostatically stable over the pH range of 3 to 12 in aqueous solution. The chemical structure of LNPs was not significantly modified compared to lignin. Meanwhile the increased content of carboxyl and aliphatic hydroxyl groups in the LNPs structure was observed. Furthermore, the thermal stability and solubility in organic solvents (ethanol, acetone and THF) of LNPs were enhanced compared to those of lignin. In vitro cell viability evaluation indicated that the prepared LNPs had no cytotoxicity and excellent biocompatibility with mouse fibroblast. Therefore, we proposed here the production of high-quality and renewable LNPs, which will provide a novel perspective for multifunctional applications of bio-based nanomaterials.

To study the long-term evolution of the transmitted CXCR4-using viruses. CCR5-using viruses (R5 viruses) predominate during primary HIV-1 infections (PHI) while CXCR4-using viruses are isolated in less than 10% of PHI.Six patients infected with an R5X4 virus, detected by a sensitive phenotypic assay during PHI, were matched with six patients infected with a pure R5 virus for sex, Fiebig stage, time of antiretroviral initiation and duration of follow-up.We used MiSeq ultra-deep sequencing to determine the composition of the virus quasispecies during PHI and at the end of follow-up (median time of follow-up: 12.5 years).X4 viruses were detected by genetic analysis in three of six samples from the R5X4 group, accounting for 1.3-100% of the virus quasispecies, during PHI, and in four of six samples (accounting for 6.7-100%) at the end of follow-up. No X4 virus was detected in the R5 group during PHI and in only one patient (accounting for 1.2%) at the end of follow-up. The complexity of the virus quasispecies at the stage of PHI was higher in the R5X4 group than in the R5 group. Complexity increased from PHI to the end of follow-up in the R5 group but remained stable in the R5X4 group.CXCR4-using viruses persisted in the peripheral blood mononuclear cells of several patients on suppressive antiretroviral therapy for a median duration of 12.5 years after PHI. The genetic complexity of HIV-1 evolved differently post-PHI in patients infected with R5X4 viruses from those infected with R5 viruses.

Motivation The circulating recombinant form of HIV-1 CRF02-AG is the most frequent non-B subtype in Europe. Anti-HIV therapy and pathophysiological studies on the impact of HIV-1 tropism require genotypic determination of HIV-1 tropism for non-B subtypes. But genotypic approaches based on analysis of the V3 envelope region perform poorly when used to determine the tropism of CRF02-AG. We, therefore, designed an algorithm based on information from the gp120 and gp41 ectodomain that better predicts the tropism of HIV-1 subtype CRF02-AG. Results We used a bio-statistical method to identify the genotypic determinants of CRF02-AG coreceptor use. Toulouse HIV Extended Tropism Algorithm (THETA), based on a Least Absolute Shrinkage and Selection Operator method, uses HIV envelope sequence from phenotypically characterized clones. Prediction of R5X4/X4 viruses was 86% sensitive and that of R5 viruses was 89% specific with our model. The overall accuracy of THETA was 88%, making it sufficiently reliable for predicting the tropism of subtype CRF02-AG sequences. Availability and implementation Binaries are freely available for download at https://github.com/viro-tls/THETA. It was implemented in Matlab and supported on MS Windows platform. The sequence data used in this work are available from GenBank under the accession numbers MK618182-MK618417.

Resume L’articulation temporo-mandibulaire supporte mal les charges en pression. Les dysfonctions sont d’origine multifactorielle. La reeducation est le plus souvent symptomatique; avec de frequentes recidives. Mieux identifier les etiologies et les traiter pourrait prevenir les recidives. Cette etude cas temoins (2 x 58 personnes) compare des patients qui ont consulte pour une dysfonction temporo-mandibulaire a des sujets sains. Le recueil des donnees est retrospectif. Les variables (ventilation, deglutition, mastication, para-fonctions) ont ete etudiees au moyen de regressions logistiques multivariees. Les resultats montrent une deglutition atypique, une mastication unilaterales et des parafonctions significativement plus nombreuses chez les patients dysfonctionnels. Cette difference n’est pas significative en ce qui concerne la, ventilation orale ou mixte. Un traitement en trois phases est propose : 1) decomprimer l’articulation et decontracter les muscles, 2) reduire les facteurs predisposants, 3) reprogrammer l’ouverture buccale et la position cervico-cephalique. Les effets de ce programme doivent faire l’objet d’une etude d’efficacite. Niveau de preuve 4.

The present study compares the performances of an in-house sequencing protocol developed on MiSeq, the Sanger method, and the 454 GS-FLX for detecting and quantifying drug-resistant mutations (DRMs) in the human immunodeficiency virus polymerase gene (reverse transcriptase [RT] and protease [PR]). MiSeq sequencing identified all the resistance mutations detected by bulk sequencing (n = 84). Both the MiSeq and 454 GS-FLX platforms identified 67 DRMs in the RT and PR regions, but a further 25 DRMs were identified by only one or other of them. Pearson's analysis showed good concordance between the percentage of drug-resistant variants determined by MiSeq and 454 GS-FLX sequencing (ρ = .77, P < .0001). The MiSeq platform is as accurate as the 454 GS-FLX Roche system for determining RT and PR DRMs and could be used for monitoring human immunodeficiency virus type 1 drug resistance.

Objectives To evaluate the diagnostic performance of the Vela next-generation sequencing (NGS) system in conjunction with the Sentosa SQ HIV Genotyping Assay for genotyping HIV-1. Methods Plasma RNA was extracted and templates prepared with the Sentosa SX instrument before sequencing the HIV-1 polymerase on the Sentosa SQ301 Sequencer (PGM IonTorrent). The Vela NGS System was compared with direct sequencing and the 454 GS-FLX (Roche) and MiSeq (Illumina) systems for genotypic resistance testing on clinical samples. Results The Vela NGS system detected majority resistance mutations in subtype B and CRF02-AG samples at 500 copies/mL and minority variants with a sensitivity of 5% at 100 000 copies/mL. The Vela NGS system and direct sequencing identified resistance mutations with 97% concordance in 46 clinical samples. Vela identified 1/20 of the 1%-5% mutations identified by 454, 5/12 of the 5%-20% mutations and 60/61 of the >20% mutations. Vela identified 3/14 of the 1%-5% mutations identified by MiSeq, 0/2 of the 5%-20% mutations and 47/47 of the >20% mutations. The resistance mutation quantifications by Vela and 454 were concordant (bias: 2.31%), as were those by Vela and MiSeq (bias: 1.06%). Conclusions The Vela NGS system provides automated nucleic acid extraction, PCR reagent distribution, library preparation and bioinformatics analysis. The analytical performance was very good when compared with direct sequencing, but was less sensitive than two other NGS platforms for detecting minority variants.

The ilama tree (Annona diversifolia Saff.) is an underutilized fruit species from Mesoamerica with a noteworthy variability in southern Mexico. The present study describes differences in knowledge and appreciation of the fruit among inhabitants from different communities in Southern Mexico, as well as differences among their ilama trees. The study took place in Tierra Caliente, a region located in the states of Guerrero and the State of Mexico. Interviews were administered in three different communities as an attempt to associate the natural morphologic variation of plants and locals’ knowledge and use of the tree in order to identify their implications for conservation. Data were analyzed by multivariate analysis, and the results suggest that the morphological variation of the plants and the commercial importance of ilama are well known by locals. Indigenous community inhabitants prefer and select only white pulp trees, whereas mestizo communities promote and conserve plants with white, red, purple, and blue pulp. Germplasm flow is dynamic in the region, and ilama fruit represents a different economic value for each community. Selling the fruit is the primary goal, but not specifically its conservation and promotion. Implications for in situ conservation of ilama in Mexico are discussed.

Starting resuscitation before clamping the umbilical cord at birth may progressively increase pulmonary blood flow while umbilical venous blood flow is still contributing to maintenance of oxygenation and left ventricle preload.To evaluate the feasibility, safety, and effects of intact cord resuscitation (ICR) on cardiorespiratory adaptation at birth in newborn infants with CDH.Prospective, observational, single-center pilot study.Physiologic variables and outcomes were collected prospectively in 40 consecutive newborn infants with an antenatal diagnosis of isolated CDH.Infants were managed with immediate cord clamping (ICC group) from 1/2012 to 5/2014 or the cord was clamped after initiation of resuscitation maneuvers (ICR group) from 6/2014 to 4/2016 (20 in each group). Ante- and postnatal markers of CDH severity were similar between groups. Resuscitation before cord clamping was possible for all infants in the ICR group. No increase in maternal or neonatal adverse events was observed during the period of ICR. The pH was higher and the plasma lactate concentration was significantly lower at one hour after birth in the ICR than in the ICC group (pH=7.17±0.1 vs 7.08±0.2; lactate=3.6±2.3 vs 6.6±4.3mmol/l, p0.05). Mean blood pressure was significantly higher in the ICR than in the ICC group at H1 (52±7.7 vs 42±7.5mmHg), H6 (47±3.9 vs 40±5.6mmHg) and H12 (44±2.9 vs 39±3.3mmHg) (p0.05).Commencing resuscitation and initiating ventilation while the infant is still attached to the placenta is feasible in infants with CDH. The procedure may support the cardiorespiratory transition at birth in infants with CDH.

International audience; Background: Patients on antiretroviral therapy could benefit from HIV-1 DNA resistance genotyping for exploring virological failure with low viral load or to guide treatment simplification. Few new generation sequencing data are available.Objective: To check that the automated deep sequencing Sentosa platform (Vela DX) detected minority resistant variants well enough for HIV DNA genotyping.Study design: We evaluated the Sentosa SQ HIV genotyping assay with automated extraction on 40 DNA longitudinal samples from treatment-experienced patients by comparison with Sanger sequencing. HIV drug resistance was interpreted using the ANRS algorithm (v29) at the threshold of 20 % and 3 %.Results: The Sentosa SQ HIV genotyping assay was 100 % successful to amplify and sequence PR and RT and 86 % to amplify and sequence IN when the HIV DNA load was >2.5 log copies/million cells. The Sentosa and Sanger sequencing were concordant for predicting PR-RT resistance at the threshold of 20 % in 14/18 samples successfully sequenced. A higher level of resistance was predicted by Sentosa in three samples and by Sanger in one sample. The prevalence of resistance was 7 % to PI, 59 % to NRTI, 31 % to NNRTI and 20 % to integrase inhibitors using the Sentosa SQ genotyping assay at the threshold of 3 %. Seven additional mutations

The ultrasound (US) assisted extraction of bio-compounds from different fruit skins (apples, bananas and persimmons) was studied. The aqueous suspensions of skins were treated by US with different energy inputs (0.033–0.299 kW·h/kg) and total time of aqueous extraction was up to 2700 s. The ionic, Zi, and total polyphenol, Zp, extraction indexes of the liquid extracts were analyzed. From microscopic images the cell wall disintegration index, Zm, was determined. Increase in US energy input caused the increase of values of Zi, Zp and Zm. The correlations between extraction parameters and the disintegration index, Zm, were discussed.

A polyol method was used to obtain ultrasmall ZnO nanoparticles (NPs) doped with iron ions and coated with a low molecular weight fucoidan in order to perform in vivo MR and ex vivo fluorescence imaging of athrothrombosis. During the synthesis, the early elimination of water by azeotropic distillation with toluene allowed us to produce NPs which size, determined by XRD and TEM, decreased from 7 nm to 4 nm with the increase of iron/zinc ratios from 0.05 to 0.50 respectively. For the highest iron content (NP-0.50) NPs were evidenced as a mixture of nanocrystals made of wurtzite and cubic phase with a molar ratio of 2.57:1, although it was not possible to distinguish one from the other by TEM. NP-0.50 were superparamagnetic and exhibited a large emission spectrum at 470 nm when excited at 370 nm. After surface functionalization of NP-0.50 with fucoidan (fuco-0.50), the hydrodynamic size in the physiological medium was 162.0 &plusmn, 0.4 nm, with a corresponding negative zeta potential of &minus, 48.7 &plusmn, 0.4 mV, respectively. The coating was evidenced by FT-IR spectra and thermogravimetric analysis. Aqueous suspensions of fuco-0.50 revealed high transverse proton relaxivities (T2) with an r2 value of 173.5 mM&minus, 1 s&minus, 1 (300 K, 7.0 T) and remained stable for more than 3 months in water or in phosphate buffer saline without evolution of the hydrodynamic size and size distribution. No cytotoxic effect was observed on human endothelial cells up to 48 h with these NPs at a dose of 0.1 mg/mL. After injection into a rat model of atherothrombosis, MR imaging allowed the localization of diseased areas and the subsequent fluorescence imaging of thrombus on tissue slices.

Hepatitis E virus (HEV) can lead to chronic infection in solid-organ transplant patients. Ribavirin is efficient for treatment of chronically infected patients. Recently, the1634R mutation in the HEV polymerase has been associated with treatment failure. However, it is unclear if this mutation can be used as a prognostic marker of treatment outcome. We studied the prevalence of the 1634R mutation in the HEV polymerase of patients starting ribavirin therapy, the influence of the 1634R variants on the viral response, the frequency of the 1634R mutation in patients whose treatment failed, and its impact on ribavirin retreatment. We analyzed pretreatment samples from 63 solid-organ transplant patients with chronic hepatitis E using deep sequencing; 42 patients had a sustained virologic response (SVR), and 21 were non-SVR patients. We detected the 1634R variant by deep sequencing in 36.5% (23/63) of the patients (proportions, 1.3 to 100%). The 1634R variant was detected in 31.0% (13/42) of baseline plasma samples from patients with SVR and in 47.6% (10/21) in the other patients ( P = 0.2). The presence of this mutation did not influence the initial decrease in viral RNA. Lastly, a second prolonged ribavirin treatment led to SVR in 70% of the patients who initially did not have SVR, despite the presence of the 1634R variant. We conclude that the presence of the 1634R variant at ribavirin initiation does not lead to absolute ribavirin resistance. Although its proportion increased in patients whose treatment failed, the presence of the 1634R variant did not compromise the response to a second ribavirin treatment.

International audience; The framework of this work was to develop an emulsion-based edible film based on a chitosan nanoparticle matrix with cellulose nanocrystals (CNCs) as a stabilizer and reinforcement filler. The chitosan nanoparticles were synthesized based on ionic cross-linking with sodium tripolyphosphate and glycerol as a plasticizer. The emulsified film was prepared through a combination system of Pickering emulsification and water evaporation. The oil-in-water emulsion was prepared by dispersing beeswax into an aqueous colloidal suspension of chitosan nanoparticles using high-speed homogenizer at room temperature. Various properties were characterized, including surface morphology, stability, water vapor barrier, mechanical properties, compatibility, and thermal behaviour. Experimental results established that CNCs and glycerol improve the homogeneity and stability of the beeswax dispersed droplets in the emulsion system which promotes the water-resistant properties but deteriorates the film strength at the same time. When incorporating 2.5% w/w CNCs, the tensile strength of the composite film reached the maximum value, 74.9 MPa, which was 32.5% higher than that of the pure chitosan film, while the optimum one was at 62.5 MPa, and was obtained by the addition of 25% w/w beeswax. All film characterizations demonstrated that the interaction between CNCs and chitosan molecules improved their physical and thermal properties.

Author(s): Dimeglio, Chloe; Raymond, Stephanie; Nicot, Florence; Jeanne, Nicolas; Carcenac, Romain; Lefebvre, Caroline; Izopet, Jacques; French National Agency for Research on AIDS and Viral Hepatitis (ANRS) AC11 Resistance Study Group | Abstract: ObjectivesTo determine how the load of rilpivirine-resistant variants (mutational load) influences the virological response (VR) of HIV-1-infected patients to a rilpivirine-based first-line regimen.Patients and methodsFour hundred and eighty-nine patients infected with HIV-1 whose reverse transcriptase gene had been successfully resistance genotyped using next-generation sequencing were given a first-line regimen containing rilpivirine. Variables associated with the VR at 12 months were identified using a logistic model. The results were used to build a multivariate model for each mutational load threshold and the R2 variations were analysed to identify the mutational load threshold that best predicted the VR.ResultsThe mutational load at baseline was the only variable linked to the VR at 12 months (P l 0.01). The VR at 12 months decreased from 96.9% to 83.4% when the mutational load was g1700 copies/mL and to 50% when the mutational load was g 9000 copies/mL. The threshold of 9000 copies/mL was associated with the VR at 12 months with an OR of 36.7 (95% CI 4.7-285.1). The threshold of 1700 copies/mL was associated with the VR at 12 months with an OR of 7.2 (95% CI 1.4-36.8).ConclusionsThere is quantifiable evidence that determining a mutational load threshold can be used to identify those patients on a first-line regimen containing rilpivirine who are at risk of virological failure. The clinical management of HIV-infected patients can be improved by evaluating the frequency of mutant variants at a threshold of l 20% together with the plasma HIV-1 viral load at the time of resistance genotyping.

International audience; Hepatitis E virus genotype 3 (HEV-3) can lead to chronic infection in immunocompromised patients, and ribavirin is the treatment of choice. Recently, mutations in the polymerase gene have been associated with ribavirin failure but their frequency before treatment according to HEV-3 subtypes has not been studied on a large data set. We used single-molecule real-time sequencing technology to sequence 115 new complete genomes of HEV-3 infecting French patients. We analyzed phylogenetic relationships, the length of the polyproline region, and mutations in the HEV polymerase gene. Eighty-five (74%) were in the clade HEV-3efg, 28 (24%) in HEV-3chi clade, and 2 (2%) in HEV-3ra clade. Using automated partitioning of maximum likelihood phylogenetic trees, complete genomes were classified into subtypes. Polyproline region length differs within HEV-3 clades (from 189 to 315 nt). Investigating mutations in the polymerase gene, distinct polymorphisms between HEV-3 subtypes were found (G1634R in 95% of HEV-3e, G1634K in 56% of HEV-3ra, and V1479I in all HEV-3efg, clade HEV-3ra, and HEV-3k strains). Subtype-specific polymorphisms in the HEV-3 polymerase have been identified. Our study provides new complete genome sequences of HEV-3 that could be useful for comparing strains circulating in humans and the animal reservoir.

The present study compares the performances of an in-house sequencing protocol developed on MiSeq, the Sanger method, and the 454 GS-FLX for detecting and quantifying drug-resistant mutations (DRMs) in the human immunodeficiency virus polymerase gene (reverse transcriptase [RT] and protease [PR]). MiSeq sequencing identified all the resistance mutations detected by bulk sequencing (n = 84). Both the MiSeq and 454 GS-FLX platforms identified 67 DRMs in the RT and PR regions, but a further 25 DRMs were identified by only one or other of them. Pearson's analysis showed good concordance between the percentage of drug-resistant variants determined by MiSeq and 454 GS-FLX sequencing (ρ = .77, P .0001). The MiSeq platform is as accurate as the 454 GS-FLX Roche system for determining RT and PR DRMs and could be used for monitoring human immunodeficiency virus type 1 drug resistance.

La ville de Vaison-la-Romaine offre une étude à la fois riche et diversifiée lorsque l’on s’interroge sur le réemploi en architecture. À la suite de la conquête romaine qui prend fin au cours de la première moitié du Ier siècle avant notre ère, la ville est implantée sur la rive droite d’un cours d’eau appelé Ouvèze. Elle est abandonnée au Moyen-Âge par la population qui migre sur la rive gauche où se développe un bourg médiéval. À l’époque moderne, la ville s’établie de nouveau sur la rive droite, là où avait été fondée la ville romaine, et s’y installe définitivement. Les diverses constructions qui se sont succédées de part et d’autre de l’Ouvèze, au cours des différentes époques, ont conservé des éléments réemployés qui constituent les précieux témoins de cette pratique. Les trois cas d’étude qui sont abordés se différencient par leur nature et leur chronologie mais ils attestent de l’usage d’une même technique pour leur mise en œuvre. Ce phénomène du réemploi invite à s’interroger sur son processus et ses raisons, ainsi que sur les éléments réemployés, leur nouvelle destination et leur fonction, mais également leur provenance. The city of Vaison-la-Romaine offers a study both rich and diversified when talking about the re-employment in architecture. The city is located on the right bank of a river called Ouvèze, following the Roman conquest, which ended in the first half of the first century BC. It was abandoned in the Middle-Ages by the population who migrated to the left of the river bank where a medieval village developed. In modern times, the city once again established itself on the right bank, where the Roman city had been founded, and settled there definitively. The various constructions which succeeded during the different periods on each side of the Ouvèze, have preserved re-employed elements which constitute the precious witnesses of this practice. The three case studies which are talked about are differentiated by their nature and chronology but they attest to the use of the same technique for their implementation. This phenomenon of re-employment invites us to question its process and its reasons as well as the re-employed elements, their new destination and their function, but also their origin.

The coreceptor used by HIV-1 must be determined before a CCR5 antagonist, part of the arsenal of antiretroviral drugs, is prescribed because viruses that enter cells using the CXCR4 coreceptor are responsible for treatment failure. HIV-1 tropism is also correlated with disease progression and so must be determined for virological studies. Tropism can be determined by next-generation sequencing (NGS), but not all of these new technologies have been fully validated for use in clinical practice. The Illumina NGS technology is used in many laboratories but its ability to predict HIV-1 tropism has not been evaluated while the 454 GS-Junior (Roche) is used for routine diagnosis. The genotypic prediction of HIV-1 tropism is based on sequencing the V3 region and interpreting the results with an appropriate algorithm. We compared the performances of the MiSeq (Illumina) and 454 GS-Junior (Roche) systems with a reference phenotypic assay. We used clinical samples for the NGS tropism predictions and assessed their ability to quantify CXCR4-using variants. The data show that the Illumina platform can be used to detect minor CXCR4-using variants in clinical practice but technical optimization are needed to improve quantification.

Less invasive surfactant therapies (LIST) use surfactant instillation through a thin tracheal catheter in spontaneously breathing infants. This review and meta-analysis investigates respiratory outcomes for preterm infants with respiratory distress syndrome treated with LIST rather than administration of surfactant through an endotracheal tube. Randomised controlled trial (RCT) full texts provided outcome data for bronchopulmonary dysplasia (BPD), death or BPD, early CPAP failure, invasive ventilation requirements and usual neonatal morbidities. Relative risks (RR) from pooled data, with subgroup analyses, were obtained from a Mantel-Haenszel analysis using a random effect model. Six RCTs evaluated LIST: 4 vs InSurE and 1 each vs delayed or immediate intubation for surfactant. LIST resulted in decreased risks of BPD (RR = 0.71 [0.52-0.99]; NNT = 21), death or BPD (RR = 0.74 [0.58-0.94]; NNT = 15) and early CPAP failure or invasive ventilation requirements (RR = 0.67 [0.53-0.84]; NNT = 8 and RR = 0.69 [0.53-0.88]; NNT = 6). Compared to InSurE, LIST decreased the risks of BPD or death (RR = 0.63 [0.44-0.92]; NNT = 11) and of early CPAP failure (RR = 0.71 [0.53-0.96]; NNT = 11). Common neonatal morbidities were not different.Respiratory management with LIST decreases the risks of BPD and BPD or death, and the need for invasive ventilation. This strategy appears safe, but long-term follow-up is lacking. What is Known: • Initial management of preterm infants with CPAP decreases the risk of death or BPD, but many still require surfactant or invasive ventilation. • Surfactant can be instilled through a tracheal thin catheter while the infant breathes on CPAP, but improvement in BPD is inconsistent between studies. What is New: • Less invasive surfactant therapy (LIST) strategies decrease the risks of BPD, of death or BPD, and of CPAP failure compared to strategies where surfactant is administered through an endotracheal tube. • LIST strategies decrease the risks of the composite outcome of BPD or death and of early CPAP failure when compared to "intubation-surfactant-extubation" approaches.

This open-label, multicenter, phase 1B/2 trial assessed AUY922 plus trastuzumab in patients with locally advanced or metastatic HER2-positive breast cancer previously treated with chemotherapy and anti-HER2 therapy. This study was composed of a dose-escalation part with AUY922 administered weekly at escalating doses with trastuzumab 2 mg/kg/week (phase 1B), followed by a phase 2 part using the same regimen at recommended phase 2 dose (RP2D). The primary objectives were to determine the maximum tolerated dose (MTD) and/or RP2D (phase 1B), and to evaluate preliminary antitumor activity (phase 2) of AUY922 plus trastuzumab at MTD/RP2D. Forty-five patients were treated with AUY922 plus trastuzumab (4 in phase 1B with AUY922 at 55 mg/m(2) and 41 in phase 1B/2 with AUY922 at 70 mg/m(2) [ 7 in phase 1B and 34 in phase 2]). One patient in phase 1B (70 mg/m2) experienced a dose-limiting toxicity (grade 3 diarrhea); the RP2D was weekly AUY922 70 mg/m(2) plus trastuzumab. Of the 41 patients in the 70 mg/m(2) cohort, the overall response rate (complete or partial responses) was 22.0% and 48.8% patients had stable disease. Study treatment-related adverse events occurred in 97.8% of patients; of these, 31.1% were grade 3 or 4. Forty-one patients (91.1%) reported ocular events (82.3% had grade 1 or 2 events). Two patients (4.4%) had ocular events leading to the permanent discontinuation of study treatment. AUY922 at 70 mg/m(2) plus trastuzumab standard therapy is well tolerated and active in patients with HER2-positive metastatic breast cancer who progressed on trastuzumab-based therapy.

HIV-1 coreceptor usage must be accurately determined before starting CCR5 antagonist-based treatment as the presence of undetected minor CXCR4-using variants can cause subsequent virological failure. Ultra-deep pyrosequencing of HIV-1 V3 env allows to detect low levels of CXCR4-using variants that current genotypic approaches miss. However, the computation of the mass of sequence data and the need to identify true minor variants while excluding artifactual sequences generated during amplification and ultra-deep pyrosequencing is rate-limiting. Arbitrary fixed cut-offs below which minor variants are discarded are currently used but the errors generated during ultra-deep pyrosequencing are sequence-dependant rather than random. We have developed an automated processing of HIV-1 V3 env ultra-deep pyrosequencing data that uses biological filters to discard artifactual or non-functional V3 sequences followed by statistical filters to determine position-specific sensitivity thresholds, rather than arbitrary fixed cut-offs. It allows to retain authentic sequences with point mutations at V3 positions of interest and discard artifactual ones with accurate sensitivity thresholds.

The aims of this study were to assess the characteristics of three-dimensional displacement of the centre of mass of the body (CMb) during walking in healthy children and to compare it with those of young adults. Twenty-one children (11 males, 10 females; age range 1 to 9 years) were recruited from the nursery and school attached to the Université catholique de Louvain, Brussels, Belgium; and three young adults (one male two females; mean age 26 years 4 months) were recruited from the Rehabilitation and Physical Medicine Unit of the same university. Displacement of CMb was assessed at different walking velocities in the children and adults by two successive mathematical integrations of ground reaction forces, measured by a large strain-gauge force platform. Displacement of CMb was controlled for leg length of the participant to eliminate the scaling effect that is dependent on growth. Results showed that vertical and lateral amplitudes of the CMb when controlled for leg length were greater for children before 4 years of age and that the forward amplitude when controlled for leg length was greater for children before 7 years of age. We conclude that the development of mature human CMb displacement during independent walking is a gradual neural process, evolving until the age of 7 years.

Background Non alcoholic fatty liver disease (NAFLD) results from alteration in lipid synthesis and elimination mechanisms such as very-low density lipoprotein (VLDL) production and de novo lipogenesis. Persistent organic pollutants (POPs) are chemicals that were mostly used historically as pesticides, solvents, flame retardant, and other applications. Among POPs, polychlorinated biphenyls (PCB) have been recognized to be of environmental and potential toxicologic concerns. Specifically, PCB126 could act as endocrine disruptors and has recently been associated with hepatic fat accumulation. The purpose of the study was to investigate the effects of PCB126 on the molecular development of NAFLD using hepatocyte and rat models. Methods Hepatocytes were exposed to PCB 126 for 72 h and lipid accumulation in cells was quantified by Oil-Red-O. Rats were injected with a single dose of PCB126 or vehicle. Seven days later, liver triglycerides (TAG) content was measured along with protein quantification of hepatic microsomal triglyceride transfer protein (MTP), sterol regulatory element-binding protein 1c (SREBP1c) and diacylglycerol O-acyltransferase 2 (DGAT-2). Results Exposure to PCB126 resulted in significant increases of lipid accumulation in hepatocytes (38 %, P P P P P Conclusions It seems that exposure to PCB126 has an important emerging role in the pathophysiology of NAFLD by 1) altering elimination mechanisms such as VLDL synthesis and secretion, through MTP; and 2) increasing hepatic TAG synthesis mechanisms through DGAT 2 and SREBP1c.

The presence of low-frequency HIV-1 variants with mutations making them resistant to non-nucleoside reverse-transcriptase inhibitors (NNRTI) could influence the virological response to first-line NNRTI therapy.This study was designed to describe the proportions and quantities of NRTI and NNRTI-resistant variants in patients with successful first-line NNRTI therapy.We evaluated the presence of drug-resistance mutations (DRMs) prior to treatment initiation in 131 naive chronically HIV-1-infected patients initiating NNRTI-based first-line therapy. DRMs were detected by ultradeep pyrosequencing (UDPS) on a GS Junior instrument (Roche).The mean HIV RNA concentration was 4.78 ± 0.74 log copies/mL and the mean CD4 cell count was 368 ± 184 CD4 cells/mm(3). Patients were mainly infected with subtype B (68%) and 96% were treated with efavirenz. The sensitivity threshold for each mutation was 0.13-1.05% for 2000 reads. Major NRTI-resistant or NNRTI-resistant mutations were detected in 40 patients (33.6%). The median frequency of major NRTI-resistant mutations was 1.37% [IQR: 0.39-84.1], i.e.: a median of 556 copies/mL [IQR: 123-37,553]. The median frequency of major NNRTI-resistant DRMs was 0.78% [IQR: 0.67-7.06], i.e.: a median of 715 copies/mL [IQR: 391-3452]. The genotypic susceptibility score (GSS) of 9 (7.3%) patients with mutations to given treatment detected by UDPS was 1.5 or 2.First-line NNRTI-based treatment can produce virological success in naïve HIV-1-infected patients harboring low-frequency DRMs representing1% of the viral quasispecies. Further studies are needed to determine the clinical cut-off of low-frequency resistant variants associated to virological failure.

The concept of activation in the absence of agonists has been demonstrated for many GPCRs and is now solidified as one of the principal aspects of GPCR signaling. In this chapter, we review how dopamine receptors demonstrate this ability. Although difficult to prove in vivo due to the presence of endogenous dopamine and lack of subtype-selective inverse agonists and "pure" antagonists (neutral ligands), in vitro assays such as measuring intracellular cAMP, [(35)S]GTPγS binding, and [(3)H]thymidine incorporation have uncovered the constitutive activation of D1- and D2-class receptors. Nevertheless, because of limited and inconsistent findings, the existence of constitutive activity for D2-class receptors is currently not well established. Mutagenesis studies have shown that basal signaling, notably by D1-class receptors, is governed by the collective contributions of transmembrane domains and extracellular/intracellular loops, such as the third extracellular loop, the third intracellular loop, and C-terminal tail. Furthermore, constitutive activities of D1-class receptors are subjected to regulation by kinases. Among the dopamine receptor family, the D5 receptor subtype exhibits a higher basal signaling and bears resemblance to constitutively active mutant forms of GPCRs. The presence of its constitutive activity in vivo and its pathophysiological relevance, with a brief mention of other subtypes, are also discussed.

8034 Background: Identifying molecular drivers in lung squamous cell carcinoma (SCC) is a major medical need since no targeted therapy is available yet for this subtype. Fibroblast growth factor receptors (FGFRs) are physiologically involved in cell proliferation and survival. Genetic alteration of FGFR genes leads to deregulated activation in various cancers, and FGFR1 amplification has been detected in lung SCC. Here, we report on patients (pts) with FGFR1-amplified lung SCC treated in a phase 1 study of BGJ398, a potent, selective pan-FGFR inhibitor. Methods: While this phase 1 dose-escalation study enrolled pts ≥ 18 years of age with any FGFR genetically altered tumor, we report here on the subgroup of pts with FGFR1-amplified advanced or metastatic lung SCC treated with 100 or 125 mg BGJ398 once daily in 28-day cycles. Pts who had progressed following at least 1 line of therapy, including platinum, were eligible for enrollment. FGFR1-amplified tumors were identified by fluorescent/chromogenic in situ...

The concept of activation in the absence of agonists has been demonstrated for many GPCRs and is now solidified as one of the principal aspects of GPCR signaling. In this chapter, we review how dopamine receptors demonstrate this ability. Although difficult to prove in vivo due to the presence of endogenous dopamine and lack of subtype-selective inverse agonists and "pure" antagonists (neutral ligands), in vitro assays such as measuring intracellular cAMP, [(35)S]GTPγS binding, and [(3)H]thymidine incorporation have uncovered the constitutive activation of D1- and D2-class receptors. Nevertheless, because of limited and inconsistent findings, the existence of constitutive activity for D2-class receptors is currently not well established. Mutagenesis studies have shown that basal signaling, notably by D1-class receptors, is governed by the collective contributions of transmembrane domains and extracellular/intracellular loops, such as the third extracellular loop, the third intracellular loop, and C-terminal tail. Furthermore, constitutive activities of D1-class receptors are subjected to regulation by kinases. Among the dopamine receptor family, the D5 receptor subtype exhibits a higher basal signaling and bears resemblance to constitutively active mutant forms of GPCRs. The presence of its constitutive activity in vivo and its pathophysiological relevance, with a brief mention of other subtypes, are also discussed.

Background: Fibroblast growth factor receptors (FGFRs) play a role in cell proliferation and survival. Genetic alterations of FGFRs can lead to deregulated activation in various cancers, including squamous cell carcinoma (SCC) of the lung and urothelial bladder cancer. Here, we report on a phase I study of BGJ398 a potent, selective pan-FGFR inhibitor. Methods: Eligible patients (pts; ≥ 18 years of age) had tumors with any FGFR genetic alteration identified by central or local prescreening. Pts received BGJ398 once or twice daily (qd or bid) in 28-day cycles in escalating cohorts. Dose-limiting toxicities (DLTs) were predefined and included both severe events and those resulting in significant dosing delays. Upon determination of maximum tolerated dose (MTD), 3 expansion arms were treated: arm 1 included FGFR1-amplified SCC of the lung (continuous qd dosing); arms 2 and 3 included FGFR-altered solid tumors given qd continuous doses (arm 2) or qd doses on a 3-weeks-on/1-week-off schedule (arm 3). Results: As of September 24, 2013, 94 pts were enrolled (median age 57.5 years). Pts were treated in qd dose cohorts (5-150 mg; n = 90) or a bid dose cohort (50 mg; n = 4). Arms 1 and 3 are ongoing. Of the 82 pts who discontinued, the most common reasons were disease progression (n = 56), consent withdrawal (n = 12), and adverse events (AEs; n = 10). DLTs during dose escalation were grade 3 aminotransferase increases (1 each at 100 and 125 mg), hyperphosphatemia (125 mg), and grade 1 corneal toxicity (150 mg). The 125-mg qd dose was identified as the MTD. Common treatment-emergent AEs (any grade, suspected to be related) at 125 mg qd (n = 41) include hyperphosphatemia (78%), stomatitis (37%), alopecia (32%), decreased appetite (32%), and fatigue (22%) and were generally mild, with stomatitis (7%) the only grade 3/4 AE occurring ≥ 5%. Elevated serum phosphate levels, a pharmacodynamic marker of FGFR pathway inhibition, could be managed through diet, phosphate lowering therapy, and drug interruptions. Preliminary analysis of efficacy data indicates tumor regressions in pts with various FGFR genetic alterations, including 4 of 5 pts with urothelial cell carcinomas (4 of which originated in the bladder) with FGFR3-activating mutations (with tumor reductions ranging from 27% to 48%). Additionally, 1 pt with FGFR1-amplified SCC lung cancer achieved confirmed partial response (PR) while another pt achieved a PR confirmed after 27 days. Tumor reductions were also observed in cholangiocarcinoma with an FGFR2 gene fusion, and FGFR1-amplified breast cancer. Conclusions: BGJ398 had a tolerable safety profile and demonstrated single-agent activity in pts with FGFR-genetically altered solid tumors. The 3-weeks-on/1-week-off schedule has been chosen for future studies based on its improved safety profile. Clinical activity was observed in multiple tumor types, and pts with FGFR3-mutated bladder cancer may be especially sensitive to BGJ398. Citation Format: Lecia V. Sequist, Phillippe Cassier, Andrea Varga, Josep Tabernero, Jan HM Schellens, Jean-Pierre Delord, Patricia LoRusso, D. Ross Camidge, Manuel Hidalgo Medina, Martin Schuler, Mario Campone, G. Gary Tian, Steven Wong, Jesus Corral, Randi Isaacs, Suman K. Sen, Diana Graus Porta, Swarupa G. Kulkarni, Caroline Lefebvre, Jürgen Wolf. Phase I study of BGJ398, a selective pan-FGFR inhibitor in genetically preselected advanced solid tumors. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr CT326. doi:10.1158/1538-7445.AM2014-CT326

530 Background: AUY922 is a novel HSP90 inhibitor that causes degradation of oncogenic proteins. A Phase I study in advanced solid tumors gave a recommended Phase II dose (RP2D) of 70 mg/m2. In the Phase II part of the study, single-agent AUY922 showed preliminary activity in HER2+ metastatic breast cancer (mBC) (Schröder et al ASCO 2011). This Phase Ib/II study assessed safety and efficacy of AUY922 + trastuzumab (T) in pts with T-refractory HER2+ advanced/mBC. Methods: Pts with HER2+ BC (ECOG PS ≤1) who progressed on 1–2 prior anti-HER2 regimens with at least 1 T-containing regimen, received weekly AUY922 (55 or 70 mg/m2, IV) + T (2 mg/kg) in a 28-day cycle. The Phase Ib primary objective was to define a MTD/RP2D of AUY922 combined with T using an adaptive Bayesian logistic regression model. The Phase II primary objective is to evaluate preliminary antitumor activity at this dose (ORR). Secondary objectives included evaluation of PK, safety and preliminary efficacy (PFS, OS). Results: In the Phase Ib part (11 pts) there was 1 DLT (Grade [Gr] 3 diarrhea), and the full standard single dose of AUY922 was recommended for use in combination with T. As of 11th January 2012, 31 HER2+ BC pts (median age 51 years [range 29–71]; 84% invasive ductal carcinoma; 52% estrogen receptor-positive; 81% had received ≥2 prior antineoplastic regimens) received AUY922 at 55 mg/m2 (n=5) or 70 mg/m2 (n=26) + T. The most common study drug-related AEs (all Gr) were diarrhea (25 [81%] pts), visual AEs (e.g. vision blurred, visual impairment, photopsia) (25 [81%] pts), and nausea (11 [35%] pts). Gr 3/4 AEs were rare; most frequently diarrhea and visual impairment (both 6%). Treatment was ongoing for 13 (37%) pts at the cutoff date. Preliminary Phase II best overall response data (RECIST, investigator assessed) was available for 22 pts at AUY922 70 mg/m2. Any partial responses were reported in 5/22 (23%) pts. Conclusions: The data suggests AUY922 + T is well tolerated. Common AEs were considered manageable, and eye disorders were reversible with treatment interruption, dose reduction or discontinuation. The combination of AUY922 + T is active in pts with HER2+ mBC who progressed on T-based therapy, and the regimen warrants further investigation.

Cette recherche s’intéresse aux enjeux du « bien-vieillir » de la population, en lien avec la dépendance, le soutien à domicile et la qualité de vie. Les recherches montrent que lorsqu’une personne vieillissante est dépendante, la présence d’un aidant est indispensable. Cette étude explore comment les ergothérapeutes interviennent dans la coopération avec le système familial pour favoriser le processus d’autonomisation de la personne vieillissante dépendante à domicile. Une méthode clinique a permis de mener des entretiens auprès de trois ergothérapeutes intervenant à domicile auprès de cette population. Les résultats ont été traités selon deux méthodes d’analyse de données textuelles. Les résultats des entretiens suggèrent que l’ergothérapeute est centré sur l’information et l’orientation avec des actions en prenant en compte l’aidant. Toutefois, les limites résident autour du rôle du professionnel, dans sa problématique institutionnelle, et la place des acteurs dans une approche systémique. Il serait intéressant de poursuivre cette étude par l’analyse de l’organisation et de la fonction des acteurs dans leurs interactions afin de donner du sens à l’engagement des aidants.

Introduction, dans un contexte de vieillissement croissant de la population, le secteur du Maintien à Domicile (MAD) en France constitue une perspective professionnelle majeure pour les ergothérapeutes. Ils sont amenés à travailler avec les aidants professionnels (AP) du domicile, acteurs-clés de ce secteur, la collaboration interprofessionnelle est désormais attendue par les instances de santé : les ergothérapeutes en sont-ils partie prenante ? Objectif, dans quels types de logique et de posture de collaboration s’inscrivent les ergothérapeutes du MAD auprès des AP ? Quels facteurs spécifiques peuvent les déterminer ? Matériel et Méthode, un questionnaire d’enquête basée sur une méthode différentielle a été envoyé à 40 ergothérapeutes intervenant dans le secteur du MAD, Résultats, 30 réponses ont été reçues et commencées à être traitées statistiquement, conclusion, à la mesure de notre analyse de données, les ergothérapeutes du MAD sembleraient être partie prenante pour une collaboration interprofessionnelle avec les AP.