High-pressure intrapleural chemotherapy: feasibility in the pig model.

International audienceABSTRACT: BACKGROUND: The usual treatments for pleural malignancies are mostly palliative. In contrast, peritoneal malignancies are often treated with a curative intent by cytoreductive surgery and intraperitoneal chemotherapy. As pressure has been shown to increase antitumor efficacy, we applied the concept of high-pressure intracavitary chemotherapy to the pleural space in a swine model. METHODS: Cisplatin and gemcitabine were selected because of their antineoplasic efficacy in vitro in a wide spectrum of cancer cell lines. The pleural cavity of 21 pigs was filled with saline solution; haemodynamic and respiratory parameters were monitored. The pressure was increased to 15-25 cm H2O. This treatment was associated with pneumonectomy in 6 pigs. Five pigs were treated with chemotherapy under pressure. RESULTS: The combination of gemcitabine (100 mg/l) and cisplatin (30 mg/l) was highly cytotoxic in vitro. The maximum tolerated pressure was 20 cm H20, due to haemodynamic failure. Pneumonectomy was not tolerated, either before or after pleural infusion. Five pigs survived intrapleural chemotherapy associating gemcitabine and cisplatin with 20 cm H2O pressure for 60 min. CONCLUSIONS: High-pressure intrapleural chemotherapy is feasible in pigs. Further experiments will establish the pharmacokinetics and determine whether the benefit already shown in the peritoneum is also obtained in the pleura

Performance of matrices developed to identify patients with early rheumatoid arthritis with rapid radiographic progression despite methotrexate therapy: an external validation study based on the ESPOIR cohort data

International audienceIntroduction Use of prediction matrices of risk or rapid radiographic progression (RRP) for early rheumatoid arthritis (RA) in clinical practice could help to better rationalise the first line of treatment. Before use, they must be validated in populations that have not participated in their construction. The main objective is to use the ESPOIR cohort to validate the performance of 3 matrices (ASPIRE, BEST and SONORA) to predict patients at high risk of RRP at 1 year of disease despite initial treatment with methotrexate (MTX).Methods We selected from the ESPOIR cohort 370 patients receiving MTX or leflunomide (LEF) for ≥3 months within the first year of follow-up. Patients were assessed clinically every 6 months, and structural damage progression seen on radiography was measured by the van der Heijde-modified Sharp score (vSHS) at 1 year. RRP was defined as an increase in the vSHS≥5 points during the first year.Results At 1 year, the mean vSHS score was 1.7±5.0 and 46 patients had RRP. The ASPIRE matrix had only moderate validity in the ESPOIR population, with area under the receiver operating characteristic curve (AUC) <0.7. The AUC for the BEST and SONORA matrices were 0.73 and 0.76. Presence of rheumatoid factor (RF)—or anti-citrullinated protein antibodies (ACPAs) and initial structural damage were always predictive of RRP at 1 year. Disease Activity Score in 28 joints (DAS28) and C reactive protein (ASPIRE threshold) were not associated with RRP.Conclusions Matrices to identify patients at risk of RRP tested in the ESPOIR cohort seem to perform moderately. There is no matrix that shows clearly superior performance

Tumor necrosis factor alpha and adalimumab differentially regulate CD36 expression in human monocytes

In chronic inflammatory diseases, such as rheumatoid arthritis, inflammation acts as an independent cardiovascular risk factor and the use of anti-inflammatory drugs, such as anti-tumor necrosis factor alpha (anti-TNFα), may decrease this risk. The phagocytosis of oxidized low density lipoproteins (LDLs) accumulated in the subendothelium by mononuclear cells influences atherosclerosis and depends on CD36 expression. We investigated the role of TNFα and adalimumab, a human anti-TNFα monoclonal antibody widely used in human pathology, in CD36 expression in human monocytes. Human monocytes were prepared by adherence from whole-blood buffy-coat fractions from healthy donors. CD36 expression was assessed by RT-PCR and flow cytometry, with various TNFα or adalimumab concentrations. Implication of peroxisome proliferator-activated receptor (PPAR)γ in the regulation of CD36 expression was assessed using specific inhibitor or gel shift assays. The impact of redox signaling was investigated using quantification of reactive oxygen species, antioxidant and a NADPH oxidase inhibitor. The F(ab')2 fragment of adalimumab was isolated and its effect was analyzed. TNFα inhibits both CD36 membrane expression and mRNA expression. This inhibition involves a reduction in PPARγ activation. In contrast, adalimumab increases both CD36 membrane expression and mRNA expression. This induction is independent of the Fc portion of adalimumab and involves redox signaling via NADPH oxidase activation. CD36 expression on human monocytes is inhibited by TNFα and independently increased by adalimumab. These data highlight that pro-inflammatory cytokines and their specific neutralization influence the expression of cellular receptors implicated in atherosclerosis. Further studies are needed to investigate the clinical implications of these results in accelerated atherosclerosis observed in rheumatoid arthritis

Fluorescent labeling in semi-solid medium for selection of mammalian cells secreting high-levels of recombinant proteins

<p>Abstract</p> <p>Background</p> <p>Despite the powerful impact in recent years of gene expression markers like the green fluorescent protein (GFP) to link the expression of recombinant protein for selection of high producers, there is a strong incentive to develop rapid and efficient methods for isolating mammalian cell clones secreting high levels of marker-free recombinant proteins. Recently, a method combining cell colony growth in methylcellulose-based medium with detection by a fluorescently labeled secondary antibody or antigen has shown promise for the selection of Chinese Hamster Ovary (CHO) cell lines secreting recombinant antibodies. Here we report an extension of this method referred to as fluorescent labeling in semi-solid medium (FLSSM) to detect recombinant proteins significantly smaller than antibodies, such as IGF-E5, a 25 kDa insulin-like growth factor derivative.</p> <p>Results</p> <p>CHO cell clones, expressing 300 μg/ml IGF-E5 in batch culture, were isolated more easily and quickly compared to the classic limiting dilution method. The intensity of the detected fluorescent signal was found to be proportional to the amount of IGF-E5 secreted, thus allowing the highest producers in the population to be identified and picked. CHO clones producing up to 9.5 μg/ml of Tissue-Plasminogen Activator (tPA, 67 kDa) were also generated using FLSSM. In addition, IGF-E5 high-producers were isolated from 293SF transfectants, showing that cell selection in semi-solid medium is not limited to CHO and lymphoid cells. The best positive clones were collected with a micromanipulator as well as with an automated colony picker, thus demonstrating the method's high throughput potential.</p> <p>Conclusion</p> <p>FLSSM allows rapid visualization of the high secretors from transfected pools prior to picking, thus eliminating the tedious task of screening a high number of cell isolates. Because of its rapidity and its simplicity, FLSSM is a versatile method for the screening of high producers for research and industry.</p

Perception de la parole et IRM : réalisation, évaluation et validation d'un système permettant une stimulation sonore de qualité en cours de séquence IRM

International audienceThis study describes the design and the assessment of a MRI-compatible sound production hardware. This system was developed to permit auditory studies with Magnetic Resonance Imaging (MRI) techniques. An important disadvantage caused by the MR imager is the acoustic noise generated during data acquisition, due to the fast gradient switching interacting with the main magnetic field. Several solutions were explored to reduce noise and to provide audio stimuli with a reasonable quality. The sound production system was first tested by instrumental methods (sound level, spectral analysis). Finally, perceptual tests consisting in intelligibility, semantic decision and prosodic judgement were achieved to validate the installation.Cette étude décrit la réalisation d'un dispositif de stimulation auditive pour un imageur à résonnance magnétique fonctionelle de 3 Tesla. Le plus important probléme de ces systémes réside dans l'émission d'un niveau de bruit considérable au cours de son fonctionnement qui les rend quasi impossible à utiliser pour des études en stimulation auditive.Plusieurs solutions de stimulation de réalisation locale sont proposées pour permettre de gérérer au tympan des sujet une stimulation de qualité raisonnable. Les stimulus ainsi générés sont d'abord testés au moyen de méthodes physiques classiques (analyses acoustiques). Ils sont ensuite testés au moyen de tests d'intelligibilité de décision sémantique et de jugement prosodique afin de les valider pour des études de psycho linguistique et psycho acoustique

Transcripts of ceruloplasmin but not hepcidin, both major iron metabolism genes, exhibit a decreasing pattern along portocentral axis of mouse liver

International audienc

Le plus ancien enfant d'Aquitaine : Combe-Grenal 31 (Domme, France)

Le récolement des collections du Musée national de Préhistoire a permis, lors du travail sur la faune découverte au cours des fouilles de F. Bordes à Combe-Grenal, site de référence, l’identification d’un nouveau fossile humain, Combe-Grenal 31. Il provient de la couche 60. En fonction des vestiges archéologiques et paléontologiques qu'elle livre, et de comparaisons avec des niveaux aussi anciens de différents gisements, cette couche s’est probablement formée lors du dernier tiers du stade isotopique 6. Combe-Grenal 31 correspond à une incisive inférieure droite de la dentition déciduale d’un enfant d’environ 3 ans ± 12 mois. Sa couronne présente de grandes dimensions malgré une forte attrition de la face occlusale. La courbure de la face vestibulaire, ainsi que le tubercule lingual sont bien marqués. Les crêtes marginales sont un peu saillantes. Ses caractères morphologiques et leur comparaison avec d'autres fossiles européens ainsi que l’ellipse d’équiprobabilité réalisée à partir des dimensions de la couronne nous permettent de souligner des similitudes avec les dents équivalentes d’autres enfants néandertaliens des stades isotopiques 5, 4 ou 3 et deux spécimens européens rapportés au stade 6

Le plus ancien enfant d'Aquitaine : Combe-Grenal 31 (Domme, France)

Le récolement des collections du Musée national de Préhistoire a permis, lors du travail sur la faune découverte au cours des fouilles de F. Bordes à Combe-Grenal, site de référence, l’identification d’un nouveau fossile humain, Combe-Grenal 31. Il provient de la couche 60. En fonction des vestiges archéologiques et paléontologiques qu'elle livre, et de comparaisons avec des niveaux aussi anciens de différents gisements, cette couche s’est probablement formée lors du dernier tiers du stade isotopique 6. Combe-Grenal 31 correspond à une incisive inférieure droite de la dentition déciduale d’un enfant d’environ 3 ans ± 12 mois. Sa couronne présente de grandes dimensions malgré une forte attrition de la face occlusale. La courbure de la face vestibulaire, ainsi que le tubercule lingual sont bien marqués. Les crêtes marginales sont un peu saillantes. Ses caractères morphologiques et leur comparaison avec d'autres fossiles européens ainsi que l’ellipse d’équiprobabilité réalisée à partir des dimensions de la couronne nous permettent de souligner des similitudes avec les dents équivalentes d’autres enfants néandertaliens des stades isotopiques 5, 4 ou 3 et deux spécimens européens rapportés au stade 6