Search

Introduction: Management of testicular cancer requires a complete evaluation to confirm the localized stage and effective treatment according to guidelines to ensure the best outcome. The primary objective of this study was to evaluate practices at each stage of care in patients with a localized testicular tumor. The secondary objective was to evaluate the oncological prognosis of these patients according to the modalities of care. Methods: We conducted a multicenter practice evaluation study with retrospective collection and evaluation of patient records. The study was conducted in two French departments (population pool of 2 million inhabitants) between January 1, 2010, and January 31, 2015, enabling a five-year followup of patients. Patients presenting with stage I testicular tumor according to the American Joint Committee on Cancer classification were included in the analysis. Results: A total of 226 records were analyzed; 93% of patients underwent bilateral scrotal ultrasound and 93.25% had a chest-abdomen-pelvis computed tomography scan. A total of 29.65% of patients had a preoperative tumor marker assay in accordance with guidelines; 94% of patients had a total orchiectomy, with a median time of 15 days. At the end of the followup period, 17 patients had suffered a recurrence of their disease. Providing adjuvant care in accordance with guidelines reduced the risk of recurrence in patients with a seminomatous tumor. Conclusions: Our study showed heterogeneity in compliance with guidelines for evaluation and effective treatment of patients with a localized testicular tumor. Some essential practices, such as assays of tumor markers and fertility preservation for patients over 40 years, were not well carried out. Adjuvant management of localized tumors appears to be an important predictor of recurrence.

International audience; The Diels–Alder reaction kinetics between diphenylfulvene and maleimide using mechanochemistry and in liquid state is described. This reaction was carried out in solid state using a modified vibratory ball-mill with temperature control and in solution with toluene as solvent. The effect of temperature, ball mass, material, additives, and aging reaction were studied. We reported for the first time the kinetics of a mechanochemical reaction depending on the ratio ball mass/mass of the reagent. A new kinetic model has been established that corresponds well to our experimental data and allows an estimation of the global activation energy. In solution, the retro Diels–Alder reaction was observed. The reaction was second order for the formation of endo and exo and first order for the retro Diels–Alder reaction. The grinding method shows many advantages compared to solution: shorter reaction time, total reaction, and better selectivity.

International audience; Dibenzophenazine was recently synthesized mechanochemically at a high yield. On the basis of experimental kinetics and theoretical vibronic coupling density analysis, this efficient synthesis has been expected to involve two types of reaction mechanisms. In this study, the reaction pathways, including the catalytic effect of water, are determined using density functional theory. The lowest-energy path involves stepwise formation of two C–N bonds, and a concerted path with a higher barrier is also found. The highest-energy point along the latter path is not a transition state but a higher-order saddle point. Therefore, the concerted path is not generally accepted as a reaction path. However, even a small mechanical force dissolves imaginary frequencies so that the higher-order saddle point is changed to a transition state. This mechanical dissolution of imaginary frequencies is a new feature of theoretical mechanochemistry.

Summary In order to ascertain the significance of transmembrane tumor necrosis factor (tmTNF) reverse signaling in vivo, we generated a triple transgenic mouse model (3TG, TNFR1−/−, TNFR2−/−, and tmTNFKI/KI) in which all canonical tumor necrosis factor (TNF) signaling was abolished. In bone-marrow-derived macrophages harvested from these mice, various anti-TNF biologics induced the expression of genes characteristic of alternative macrophages and also inhibited the expression of pro-inflammatory cytokines mainly through the upregulation of arginase-1. Injections of TNF inhibitors during arthritis increased pro-resolutive markers in bone marrow precursors and joint cells leading to a decrease in arthritis score. These results demonstrate that the binding of anti-TNF biologics to tmTNF results in decreased arthritis severity. Collectively, our data provide evidence for the significance of tmTNF reverse signaling in the modulation of arthritis. They suggest a complementary interpretation of anti-TNF biologics effects in the treatment of inflammatory diseases and pave the way to studies focused on new arginase-1-dependent therapeutic targets.

Summary In order to ascertain the significance of transmembrane tumor necrosis factor (tmTNF) reverse signaling in vivo, we generated a triple transgenic mouse model (3TG, TNFR1−/−, TNFR2−/−, and tmTNFKI/KI) in which all canonical tumor necrosis factor (TNF) signaling was abolished. In bone-marrow-derived macrophages harvested from these mice, various anti-TNF biologics induced the expression of genes characteristic of alternative macrophages and also inhibited the expression of pro-inflammatory cytokines mainly through the upregulation of arginase-1. Injections of TNF inhibitors during arthritis increased pro-resolutive markers in bone marrow precursors and joint cells leading to a decrease in arthritis score. These results demonstrate that the binding of anti-TNF biologics to tmTNF results in decreased arthritis severity. Collectively, our data provide evidence for the significance of tmTNF reverse signaling in the modulation of arthritis. They suggest a complementary interpretation of anti-TNF biologics effects in the treatment of inflammatory diseases and pave the way to studies focused on new arginase-1-dependent therapeutic targets., Graphical abstract, Highlights • In vivo demonstration of tmTNF reverses signaling existence • tmTNF reverse signaling induces anti-oxidative stress response • tmTNF reverse signaling induces an arginase-1-mediated anti-inflammatory response • Reverse signaling is a complementary mechanism to TNF neutralization by anti-TNF, Immunology; Cell Biology

International audience; Increased osteoclastogenesis is a common feature of bone erosion, notably in osteoporosis but also in inflammatory diseases such as rheumatoid arthritis (RA) and osteoarticular infections. Human cytomegalovirus (HCMV) infection has been described to impair monocyte differentiation into macrophages and dendritic cells. However, its effect on monocyte-derived osteoclasts is yet to be determined. We showed here that in vitro HCMV infection is associated with an inhibition of osteoclastogenesis through decreased expression of colony stimulating factor 1 receptor (CSF-1R) and RANK in monocytes, which was mediated by an upregulation of quaking I-5 protein (QKI-5), a cellular RNA-interacting protein. We found that deliberate QKI5 overexpression in the absence of HCMV infection is able to decrease CSF-1R and RANK expression, leading to osteoclastogenesis inhibition. Finally, by using lentiviral vectors in a calvarial bone erosion mouse model, we showed that QKI5 inhibits bone degradation. This work identifies QKI5 as a strong inhibitor of bone resorption. Future research will point out whether QKI5 could be a target for bone pathologies. © 2019 American Society for Bone and Mineral Research

International audience; The mechanochemical synthesis of 1,2,4-triazoles by using a planetary ball-mill is studied starting from hydralazine hydrochloride. Excellent conditions were found for achieving total conversion to intermediate hydrazones in a few minutes in the presence of pyrogenic S13 silica as grinding auxiliary. The conversion to annulated 1,2,4-triazoles was optimal with iodobenzene diacetate (IBD) when using nonphenolic hydrazones, while SeO2 was found to be efficient for phenolic compounds. The one-pot two-step synthesis was also successfully conducted for the first time leading to annulated 1,2,4-triazoles. Finally, green metrics showed the efficiency of the method in comparison to the conventional syntheses of hydrazone 10 and triazole 13.

Background Macrophages are major effector cells in inflammatory rheumatisms such as Rheumatoid arthritis (RA) and Psoriatic arthritis (PsA). Depending on their microenvironment, especially cytokines content, they can display various phenotypes or “polarizations” described as inflammatory (in the presence of IFNγ or GM-CSF) or as alternative (in the presence of IL-4 or IL-10). These cytokines involve JAK/STAT signaling pathway, and their action is expected to be inhibited by JAK inhibitors (JAKi) developed in RA and PsA. How JAKi modulate polarization of macrophage, and whether this phenomenon explains the clinical benefit in RA and PsA is not fully understood. Objectives To evaluate whether JAKi modulate in vitro polarization of monocytes derived macrophages. Methods [Cell culture] CD14+ monocytes were isolated from healthy donors and differentiated as macrophages in the presence of M-CSF. Macrophages were then polarized as inflammatory macrophages (by LPS + IFNγ, or GM-CSF), or as alternative macrophages (by IL-4, or IL-10), in the presence or not of JAKi (500 nM): Baricitinib (JAK1/JAK2 specific), Tofacitinib (JAK1/JAK2/JAK3/Tyk2 specific). [Membrane markers] Surface polarization markers were evaluated by flow cytometry. [Functional assays] Cytokines production in supernatants of macrophages were quantitated by bead-based immunoassay and by ELISA. Reactive oxygen species (ROS) production triggered by LPS and/or IFNγ was assessed by flow cytometry. Phagocytosis of E.coli bioparticles was assessed by flow cytometry. Results We analyzed 12 donors. Membrane polarization markers were: CD40/CD80/CD206 for inflammatory macrophages, CD163/CD16/CD206/CD200R for alternative macrophages. Both JAKi modulated these markers. Regarding the inflammatory polarizations, Baricitinib and Tofacitinib respectively reduced CD40 and CD206 expression. Regarding the alternative polarizations, both drugs inhibited the expression of CD163, CD206 and CD200R. At thecytokines level, both drugs did not significantly modulate the pro-inflammatory/anti-inflammatory balance in supernatants. We observed a slight increase in TNF and IL-6 resulting from LPS/NFkB pathway [1]. However, Baricitinib decreased the inflammatory IP-10, and increased IL-10 production. Both JAKi did not affect ROS production in the presence of LPS. Consistently with the absence of modulation of CD16 surface expression, Baricitinib and Tofacitinib did not affect CD16-dependent phagocytosis. Conclusion Our study shows that despite distinct JAK specificity, Tofacitinib and Baricitinib maintained surface phenotypes close to those of non-activated macrophages This finding was observed whatever was the polarizing condition, thus supporting the potential benefit of JAK inhibitors in different immune diseases. References [1] Pattison MJ, J Immunol, September15, 2012, 189 (6) 2784-2792 Disclosure of Interests Marion Magnol: None declared, Benjamin Rauwel: None declared, Souraya Sayegh: None declared, Katy Diallo: None declared, Michel Baron: None declared, Emmanuelle Lacassagne: None declared, Jean-Frederic Boyer: None declared, Adeline Ruyssen-Witrand: None declared, Arnaud Constantin: None declared, Jean-Luc Davignon Grant/research support from: Pfizer Passerelle Grant, Yannick Degboe Grant/research support from: Celgene PARTNER Fellowship

Career situation of first and presenting author Post-doctoral fellow. Introduction Anti-TNF agents are widely used in rheumatoid arthritis (RA). Their effect on inflammation results from the neutralization of soluble TNF (sTNF), but also supposedly from the induction of reverse signaling through their binding to membrane TNF (tmTNF). Despite possible clinical relevance, reverse signaling has been described only in vitro but has not been proven in vivo. Objectives In this study we aim to demonstrate for the first time the existence of tmTNF reverse signaling in vivo and its importance in anti-TNF response during arthritis. Methods Triple transgenic mouse model (3TG), KO for TNFR1/TNFR2 and KI for tmTNF, thus secreting no sTNF was developed. To analyze reverse signaling, mice were injected either with etanercept (ETA, 10 mg/kg), an anti-mouse TNF antibody (MP6-XT22, rat IgG1, 10 mg/kg) or an anti-human IL17 antibody (secukinumab, SEC, 10 mg/kg) as a control. Daily clinical evaluation of K/BxN serum induced-arthritis was performed in 3TG as well as WT mice. Polarization of bone marrow-derived macrophages (BMDM) and cytokine production from non-arthritic WT and 3TG mice under the action of anti-TNF in vitro was evaluated by RT-qPCR, CBA and ELISA. Results In vivo, the administration of anti-TNF (ETA or MP6-XT22) decreased arthritic scores in WT mice (p=0.005) as well as in 3TG mice (p Conclusions Our work provides in vivo evidence for the involvement of reverse signaling in the anti-TNF-mediated modulation of arthritis. Reverse signaling is expected to result in the modulation of macrophage polarization from an inflammatory to an alternative functional phenotype in arthritic mice. Our data prompt us to consider new interpretation of the effects of anti-TNF in the treatment of RA. Disclosure of Interest None declared.

Background Macrophages contribute to the rheumatoid arthritis (RA) pathogenesis. They can display various states of activation or “ polarization ”, characterized by distinct functions in inflammation, and reversibility depending on their environment [1]. M1 polarization corresponds to the “classical”, pro-inflammatory activation as identified in RA. M2 “alternative” polarizations display pro-resolving or wound-healing properties. Data concerning the effects of RA biological drugs (bDMARDs) on macrophage polarization are scarce. Objectives To assess in vitro modulation of macrophage polarization by RA bDMARDs. Methods Blood monocytes from 14 healthy donors were positively sorted by CD14+ magnetic selection. Macrophages were Derived from Monocytes (MDM) by 5 days of culture in the presence of MCSF, and activated or not for 24h as M1 pro-inflammatory MDM (by LPS + IFNγ) or as M2 alternative MDM (by IL10 or IL4). MDM were cultured with or without bDMARDs. We evaluated 2 anti-TNF agents (etanercept (ETA), adalimumab (ADA)), 1 anti-IL6R agent (tocilizumab (TCZ)), and 1 anti-CD20 agent (rituximab (RTX)) used as control monoclonal antibody. bDMARDs effects were assessed separately on differentiation and activation phases by flow cytometric analysis of membrane markers. Functional aspects of polarization were assessed by analysis of cytokine production in supernatants (cytometric bead array) and phagocytosis (flow cytometry). MDM cultured in the presence of bDMARDs were compared to untreated MDM by a Wilcoxon matched pairs test. Results We first validated membranous polarization markers in our culture model: CD40 and CD80 as M1 (LPS + IFNγ) markers; CD16, CD163 and CD64 as M2 (IL10) markers, CD206 and CD200R as M2 (IL4) markers. Anti-TNF significantly modulated surface polarization markers of MDM obtained after differentiation. CD40 decreased with both anti-TNFs. CD16 increased with ADA, whereas opposite effects on CD64, decrease with ETA, increase with ADA, were observed. CD206 increased with ADA. CD200R increased with both anti-TNF. We observed a significant increase in CD64 with TCZ. RTX induced no modulation except an increase in CD64. Concerning the effect of bDMARDs on M1 activation (M1 MDM) shown in Figure, anti-TNF agents induced a significant decrease in M1 markers and a significant modulation in M2 (IL10) markers. We observed a decrease in CD40 and CD80, an increase in CD16, CD163, a decrease in CD64 with ETA and an increase with ADA. TCZ induced a slight but significant decrease in CD40 and an increase in CD64. RTX only affected CD64 as observed for differentiation. In M2 (IL4) et M2 (IL10) activated MDM, anti-TNF agents induced a similar trend toward M2 (IL10) phenotype. Anti-TNF agents led to a significant decrease in TNFα, IL6, IL12 and IL10 levels measured in cell culture supernatant after M1 MDM. TCZ mainly affected IL6 and TNFα productions. No significant effect was observed with RTX. Phagocytosis was superior in M2 (IL10) and M2 (IL4) activated MDM than in M1 MDM. Anti-TNF agents but not TCZ or RTX induced an increase of phagocytosis in M1 MDM. Conclusions Anti-TNF agents upregulate M2 alternative pro-resolving markers and downregulate M1 inflammatory markers in macrophages. Our results need to be extended by transcriptional analysis and evaluated in RA patients. References Martinez FO and Gordon S, F1000Prime Reports 2014, 6:13. Disclosure of Interest Y. Degboe Grant/research support from: Societe Francaise de Rhumatologie, B. Rauwel: None declared, M. Baron: None declared, J.-F. Boyer: None declared, A. Cantagrel: None declared, A. Constantin: None declared, J.-L. Davignon: None declared

International audience; The reaction mechanism for mechanochemical synthesis of dibenzophenazine was theoretically investigated in terms of the vibronic coupling density, which describes the interactions between electrons and nuclear motions. The concept theoretically indicates experimentally observed reactive sites that cannot be explained by the conventional frontier orbital theory. The results of vibronic coupling density analysis suggested the difference between reaction mechanisms under thermal and mechanochemical conditions.

International audience; Curing is generally required to stabilize film coating from aqueous polymer dispersion. This post-coating drying step is traditionally carried out in static conditions, requiring the transfer of solid dosage forms to an oven. But, curing operation performed directly inside the coating equipment stands for an attractive industrial application. Recently, the use of various advanced physico-chemical characterization techniques i.e., X-ray micro-computed tomography, vibrational spectroscopies (near infrared and Raman) and X-ray microdiffraction, allowed new insights into the film-coating structures of dynamically cured tablets. Dynamic curing end-point was efficiently determined after 4 h. The aim of the present work was to elucidate the influence of curing conditions on film-coating structures. Results demonstrated that 24 h of static curing and 4 h of dynamic curing, both performed at 60 degrees C and ambient relative humidity, led to similar coating layers in terms of drug release properties, porosity, water content, structural rearrangement of polymer chains and crystalline distribution. Furthermore, X-ray microdiffraction measurements pointed out different crystalline coating compositions depending on sample storage time. An aging mechanism might have occur during storage, resulting in the crystallization and the upward migration of cetyl alcohol, coupled to the downward migration of crystalline sodium lauryl sulfate within the coating layer. Interestingly, this new study clearly provided further knowledge into film-coating structures after a curing step and confirmed that curing operation could be performed in dynamic conditions.

Biotherapies have revolutionized the treatment of RA. However, much work is needed to understand all the mechanisms of these biotherapies, and alternatives are needed to circumvent adverse effects and the high cost of these long-lasting treatments. In this article we outline some of the approaches we have used to target monocytes/macrophages as major components of inflammation and bone homeostasis. We also discuss how anti-TNF-α antibodies target monocytes/macrophages in the complex mechanisms contributing to inhibition of inflammation.

Background Corpus callosum agenesis (CCA) is generally diagnosed in utero. Outcome appears to be better if the malformation is isolated. The aim of this study, which is the first one with a long (10 years) and standardized follow up, was to report cognitive abilities of children with isolated CCA diagnosed prenatally. Methods We prospectively evaluated 17 children. Clinical examinations, neuropsychological tests were performed each year. School achievement and personal and familial data were collected. Results Twelve children completed the entire follow up. One child was finally considered to have associated CCA, because signs of fetal alcohol syndrome had become obvious. Of the 11 other children, three (27%) had borderline intelligence whereas the intelligence levels of eight (73%) were in the normal range, although half of these children experienced some difficulties in scholastic achievement. Neither epilepsy nor intellectual deficiency was noted and intellectual quotient scores correlated strongly with the mother's education level. Conclusion Although prenatal diagnosis of isolated CCA is reliable, false postnatal diagnoses remain possible (10–20%) even with complete prenatal screening. Outcome is mostly favorable because intelligence is within the normal range for nearly 3/4 of the children. However, they frequently have mild learning difficulties. © 2012 John Wiley & Sons, Ltd.

International audience; Many synthetic methods exist for dibenzoquinoxalines but only a few for dibenzophenazines and their aza derivatives and even less are `green'. Some dibenzophenazines and dibenzopyridoquinoxaline have been efficiently obtained with good to excellent yield by a very simple method which does not require use of solvent or catalyst. Solid phase synthesis using co-grinding presents thus many advantages in developing greener synthetic organic pathways. (C) 2011 Elsevier Ltd. All rights reserved.

International audience; The aim of this study was to perform in-line Near Infrared (NIR) measurements inside a pan coater to monitor a coating operation in real-time, by predicting the increases in mass of coating materials and coating thickness. A polymer combination of ethylcellulose/poly(vinyl-alcohol)-poly(ethylene-glycol) graft copolymer was used as functional aqueous coating. Coated tablets were sampled at regular intervals during the coating operation, then subjected to either simple and fast weighing (n = 50) or accurate and non-destructive Terahertz Pulsed Imaging (TPI) measurements (n = 3). Off-line NIR spectra analysis revealed that the coating operation could efficiently be controlled by focusing on two distinct NIR regions, related to absorption bands of ethylcellulose. Principal component analysis of in-line NIR spectra gave a clear classification of the collected coated tablets. Real-time quantitative monitoring of the coating operation was successfully performed from partial least square calibration models built using either TPI or weighing as reference method. Coating thicknesses as well as mass of coating materials used as primary values provided accurate NIR predictions. A comparison study demonstrated that both reference methods led to reliable and accurate real-time monitoring of the coating operation. This work demonstrated that in-line NIR measurements associated with multivariate analyses can be implemented to monitor in real-time a pan coating operation in order to fulfil the expectations of ICH Q8 guideline on pharmaceutical development, especially in terms of PAT control strategy and reduced end-product testing.

Item does not contain fulltext OBJECTIVE: Several lines of evidence implicate cytosolic phospholipase A(2)alpha (cPLA(2)alpha) as a critical enzyme in inflammatory disorders, including rheumatoid arthritis. Since cells from the myeloid compartment regulate local and systemic disease pathogenesis, the present study was undertaken to examine the effect of cPLA(2)alpha inhibition in experimental arthritis, using a delivery system tailored to target monocyte functions by RNA interference (RNAi). METHODS: Mice with collagen-induced arthritis (CIA) were injected intravenously with an anti-cPLA(2)alpha small interfering RNA (siRNA) sequence (siPLA2) formulated as lipoplexes with the RPR209120/DOPE cationic liposome and a carrier DNA. The clinical course of joint inflammation was assessed, and the immunologic balance was analyzed by measuring T helper cell frequencies and cytokine expression. Biodistribution studies of siRNA were also performed. RESULTS: Weekly systemic injection of siPLA2 lipoplexes significantly reduced the incidence and severity of CIA, in both preventive and curative settings, as compared with findings in control animals. Histologic scores for inflammation and cartilage damage were reduced. The clinical effect was associated with local inhibition of tumor necrosis factor alpha secretion and lower cPLA(2)alpha expression and activity. The siPLA2 lipoplexes enabled triggering of in vivo RNAi-mediated gene silencing of cPLA(2)alpha in CD11b+ cells recovered from the spleen. While the treatment had no effect on anti-type II collagen (anti-CII) antibodies, CII-specific T helper cells producing interferon-gamma, but not interleukin-17, in draining lymph node cells were decreased. CONCLUSION: Our findings indicate that systemic RNAi-mediated cPLA(2)alpha gene silencing in CD11b+ cells is effective in the treatment of CIA, and Th1 suppression is one of the potential underlying mechanisms, whereas Th17 suppression is not.

Dry coating experiments were performed by using the Hybridizer (Nara). Large host silica gel (SG) particles (d50 = 55 μm) were coated with fine invited particles of magnesium stearate (MS, d50 = 4.6 μm) for different contents of MS in the mixture. The real MS mass fraction w I obtained after mechanical treatment has been determined thanks to calibration from TGA measurements. The surface structure and morphology of MS coatings were observed using environmental scanning electron microscopy (ESEM) and atomic force microscopy (AFM) [Y. Ouabbas, A. Chamayou, L. Galet, M. Baron, J. Dodds, A.M. Danna, G. Thomas, B. Guilhot, P. Grosseau, Modification of powders properties by dry coating: some examples of process and products characteristics, Proceedings of CHISA2008, Prague, August 2008, submitted for publication; L. Galet, Y. Ouabbas, A.M. Danna, G. Thomas, P. Grosseau, M. Baron, A. Chamayou, Surface morphology analysis and AFM study of silica gel particles after mechanical dry coating with magnesium stearate, Proceedings of PSA2008, UK, September 2008, submitted for publication]. AFM has been also used to measure the adhesion forces between particles. Interaction forces between the material attached to the cantilever (magnesium stearate MS) and the surface of the composite material (silica gel SG or magnesium stearate MS) have been determined at different surface locations. For different compositions w I of the mixture MS–SG, the numeric distribution and the mean value f of the forces fH obtained for MS–SG interactions or fI for MS–MS interactions have been established and the experimental curve showing the evolution of f versus w I has been derived. Models of ordered structures have been developed, implying morphological hypotheses concerning large spherical or cylindrical host particles H and small invited spherical I. Different types of distribution of I materials onto the surface of H have been considered: for examples a discrete monolayer – or multilayers – of monosized particles I on the H surface. The coordinence of MS particles around SG particles has been estimated to calculate the free SG surface fraction through different modelling and to obtain the mean force f versus composition w I . The theoretical force values have been compared to experimental ones. The deviations have been discussed in terms of guest particle distributions on the surface of the large host particles and morphological hypotheses.

National audience; Mi nes Albi a choisi le génie pharmaceutique comme l'un de ses axes de développement, en formation comme en recherche. L'objectif était d'accompa-gner les entreprises, en formant des professionnels rompus aux sciences de l'ingénieur et initiés au domaine pharmaceutique. Pour les aspects formation, dès sa création en 1992, l'École des mines d'Albi a lancé une option de dernière année destinée à former les futurs ingénieurs de l'industrie pharmaceutique au génie pharmaceutique. Y figurait un volet d'initiation à la réglementation et au langage pharmaceutique. Puis, en 1999, une première convention de parte-nariat a été signée entre l'université de Tours et Mines-Albi, pour la formation de pharmacien et ingénieur par la voie étudiante, qui permet aux étudiants pharmaciens en cours d'études (5 e année validée) de devenir également ingénieurs en deux ans. Cette possibilité est notamment proposée dans deux options de spécialisation bio-santé-ingénierie (BSI) et génie industriel (Gipsi). La figure ci-des-sous présente les principaux blocs de compétences des étudiants suivant l'option bio-santé-ingénierie proposée en dernière année (niveau M2). Cette formation a été rapidement plébiscitée par les industriels. Les conventions de partenariat ont été étendues dans un premier temps aux universités comme Paris-Sud (Paris-Saclay), Paris-Descartes, Marseille, Montpellier, Bordeaux ou encore Rennes, et aujourd'hui à la quasi-totalité des universités de L. Galet, M. Baron Mi nes Albi has chosen pharmaceutical engineering as one of its lines of development in both education and research. The aim was to support companies by creating professionals with engineering science expertise and initial experience of the pharmaceutical industry. When it was founded in 1992, Mines Albi launched a final-year elective designed to train future engineers in pharmaceutical engineering. It included an introduction to pharmaceutical regulation and terminology. Then, in 1999, a first partnership agreement was signed between the University of Tours and Mines Albi to create a pharmacy and engineering study programme, which enables current pharmacy students (post-fifth year) to become engineers in two years. This possibility is provided via two specialist electives, bio-health-engineering (BSI) and industrial engineering (Gipsi). The figure below presents the main skill sets of students taking the bio-health-engineering final-year elective (master' s degree level). The programme was warmly welcomed by manufacturers. The partnership agreements were initially extended to universities such as Paris-Sud (Paris-Saclay), Paris-Descartes, Marseille, Montpellier, Bordeaux and Rennes, and then to almost all universities in France as well as the University of

International audience; Measuring the action of energy on matter is a complex problem, especially in the case of granular materials. For example, particle size reduction by grinding generally shows poor overall energetic yields and a significant challenge in this area is to accurately estimate the energy consumed, including that stored in the particles. Fine or ultra-fine grinding processes require a lot of energy, part of which becomes internal energy and can lead to mechanochemical reactions and useful products. We studied the appearance of free radicals during the grinding of a-lactose monohydrate by means of electron spin resonance (ESR). These radicals are the same as those induced by gamma-radiation and comparison of ESR spectra intensities with those from ground products allows the determination of an `equivalent gamma-irradiation dose'. This gives a novel concept for characterizing the action of mechanical energy on matter in fine grinding by using molecular probes. This is the first example of the investigation of mechanochemical energy during the fine grinding process.

International audience; The aim of this study was to enhance the low solubility of eflucimibe, a new chemical entity which can be used as medicine to treat dyslipidemiae, by complexation with gamma-cyclodextrin. The complex was prepared using kneading method. The interaction evolution was studied during process by comparison of the semi-solid and physico-chemical states of the product. The evolution of the semi-solid state was followed by torque measurement when the evolution of physico-chemical state was studied by differential scanning calorimetry, infrared spectroscopy and by determination of the drug solubilisation profile. The interaction, which occurs during the process, is characterised by a modification of the product consistency and by a disappearance of the drug endothermic peaks, a disappearance of a drug spectral band and a widely improvement of the drug solubilisation profile. Indeed, after complete interaction, the drug quantity solubilised in specific conditions increased about 44-fold compared to those of untreated drug. Moreover, the comparison of the physico-chemical and semi-solid states during kneading process clearly shows that when the interaction takes place, a solidification of the paste occurs. The results of these works suggested that the formation of new solid phase allows an enhancement of the solubility of eflucimibe.

Infections by human CMV are controlled by cellular immune responses. Professional APC such as monocytes and macrophages can be infected in vivo and are considered as a reservoir of virus. However, CMV-specific CD4+ responses against infected APC have not been reported. To develop a model of CD4-infected APC interaction, we have transfected the U373MG astrocytoma cell line with the class II transactivator (CIITA). Confocal microscopy experiments showed that U373MG-CIITA cells expressed markers characteristic of APC. Functional assays demonstrated that infected U373MG-CIITA APC processed and presented both exogenous and endogenously neosynthesized nuclear immediate early (IE) protein 1 through the MHC class II pathway. More importantly, endogenous presentation of IE1 by infected APC lead to efficient control of CMV infection as revealed by decreased viral titer. Thus, these results describe the endogenous presentation of a nuclear viral protein by the MHC class II pathway and suggest that IE1-specific CD4+ T cells may play an important role in CMV infection by directly acting against infected APC.

La densité énergétique et protéique est l’attente principale des fabricants d’aliments composés vis-à-vis de l’amélioration qualitative des matières premières rentrant dans les formulations. À cette attente, l’amélioration des plantes peut proposer des réponses : réduction des teneurs en fibres peu ou pas digérées par les monogastriques, accroissement des taux en protéines et en acides aminés indispensables. L’amélioration de la digestibilité du phosphore est également travaillée depuis quelques années. L’objectif est essentiellement environnemental : éviter l’excrétion dans le milieu extérieur de trop grandes quantités de phosphore et les conséquences sur la qualité des eaux. Pour les acides gras, les ratios mono/poly-insaturés peuvent être modifiés.

The prodrug activation system formed by the E. coli codA gene encoding cytosine deaminase (CD) and 5-fluorocytosine (5-FC) developed for selective cancer chemotherapy suffers from a sensitivity limitation in many tumour cells. In an attempt to improve the CD/5-FC suicide association, we combined the E. coli upp gene encoding uracil phosphoribosyltransferase (UPRT) with codA gene to create the situation prevailing in E. coli, a bacterium very efficient in metabolising 5-FC. The constitutive expression of the two genes cloned on an E. coli-animal cell shuttle plasmid either in a linked or in a fused configuration was evaluated in E. coli strains selected and engineered to mimic the 5-FC metabolism encountered in mammalian cells. The simultaneous expression of codA and upp genes generated a cooperative effect resulting in a dramatic increase in 5-FC sensitivity of cells compared to the expression of codA alone. Furthermore, it was shown that the association of UPRT with CD facilitated the uptake of 5-FC, in the situation where the drug penetrates cells by passive diffusion as in mammalian cells, by directly channeling 5-fluorouracil, the product of CD, to 5-fluoroUMP, the product of UPRT.

Cell extracts of Trichoderma reesei exhibited dibasic endopeptidase activity toward the carboxylic side of KR, RR, and PR sequences. This activity was stimulated by the presence of Ca 2+ ions and localized in vesicles of low bouyant density; it therefore exhibited some similarity to yeast Kex2. Analytical chromatofocusing revealed a single peak of activity. The dibasic endopeptidase activity was strongly and irreversibly inhibited in vitro as well as in vivo by 1 mM p -amidinophenylmethylsulfonyl fluoride (pAPMSF) but not by PMSF at concentrations up to 5 mM. We therefore used pAPMSF to study the role of the dibasic endopeptidase in the secretion of protein by T. reesei . Secretion of xylanase I (proprotein processing sequence -R-R-↓-R-↓-A-) and xylanase II (-K-R-↓-Q-) was strongly inhibited by 1 mM pAPMSF, and a larger, unprocessed enzyme form was detected intracellularly under these conditions. Secretion of cellobiohydrolase II (CBH II; -E-R-↓-Q-) was only slightly inhibited by pAPMSF, and no accumulation of unprocessed precursors was detected. In contrast, secretion of CBH I (-R-A-↓-Q-) was stimulated by pAPMSF addition, and a simultaneous decrease in the concentration of intracellular CBH I was detected. Similar experiments were also carried out with a single heterologous protein, ShBLE, the phleomycin-binding protein from Streptoalloteichus hindustanus , fused to a series of model proprotein-processing sequences downstream of the expression signals of the Aspergillus nidulans gpdA promoter. Consistent with the results obtained with homologous proteins, pAPMSF inhibited the secretion of ShBLE with fusions containing dibasic (RK and KR) target sequences, but it even stimulated secretion in fusions to LR, NHA, and EHA target sequences. Addition of 5 mM PMSF, a nonspecific inhibitor of serine protease, nonspecifically inhibited the secretion of heterologous proteins from fusions bearing the NHA and LR targets. These data point to the existence of different endoproteolytic proprotein processing enzymes in T. reesei and demonstrate that dibasic processing is obligatory for the secretion of the proproteins containing this target.

International audience; A series of hydrazones, as potential therapeutic agents, were successfully synthesized in a vibratory ball-mill from various substituted organic hydrazines and phenol aldehydes. The degree of conversion was increased by high electronic density on the amino group of the hydrazine reactant, as well as low steric hindrance around both reactive sites. In this particular case, the flexibility of the chain bearing the amino reactive site of hydrazine was highlighted as a factor influencing the reaction rate. The results showed that hydrazones could be obtained with more than a 99% transformation, without concurring by-products. This is highly adapted to the synthesis of active pharmaceutical ingredients, requiring a high level of purity. Owing to the fact that neither an environmentally unadvisable reagent nor additives or catalysts were added to achieve the transformation, this synthesis provides a good example and a prefiguration of an efficient green pharmaceutical process.

Issu de : 3rd International conference on particulate processes in the pharmaceutical industry, Gold Coast, AUSTRALIA, 24–29 July 2011; International audience; The mechanism of the solvent-free solid-state dibenzophenazine synthesis by dry co-grinding in a vibratory ball-mill has been investigated. The kinetics of the transformation was followed by HPLC and granulometry evolutions were quantified after co-grinding. The mechanism assumed involves a quinone imine intermediate formed during the first step of the reaction (addition of an amino group to a carbonyle) which is favored by the orbital overlaps between reagents. A water molecule formation occurs during the following step and hydrogen bonds are formed: the water molecule forms a bridge between the reactive centers of the quinone imine, and acts as a catalyst for the completion of the reaction. A push-pull mechanism involving the water bridge is proposed: the energy barrier is reduced by this way. Finally, two thermodynamic drivers favor the dibenzophenazine formation: the increased aromacity number in the product and the stabilization, thanks to water molecules.; L'article est une approche préliminaire du mécanisme de la synthèse mécanochimique de la dibenzophenazine. L'intervention d'une molécule d'eau est proposée, qui pourrait agir comme catalyseur vert dans une phase statique succédant à l'activation mécanochimique. Ces travaux seront complétés ultérieurement par l'étude théorique du mécanisme par couplage vibronique et par une étude cinétique du mécanisme en phase dynamique d'activation mécanochimique, ainsi qu'en phase statique.

International audience; Scaling-up the extrusion-spheronization process involves the separate scale-up of each of the five process steps: dry mixing, granulation, extrusion, spheronization, and drying. The aim of the study was to compare two screw extrusion systems regarding their suitability for scaling-up. Two drug substances of high-and low-solubility in water were retained at different concentrations as formulation variables. Different spheronization times were tested. The productivity of the process was followed up using the extrusion rate and yield. Pellets were characterized by their size and shape, and by their structural and mechanical properties. A response surface design of experiments was built to evaluate the influence of the different variables and their interactions on each response, and to select the type of extrusion which provides the best results in terms of product quality, the one which shows less influence on the product after scale-up (''scalability'') and when the formula used changes (''robustness''), and the one which allows the possibility to adjust pellet properties with spheronization variables (''flexibility''). Axial system showed the best characteristics in terms of product quality at lab and industrial scales, the best robustness at industrial scale, and the best scalability, by comparison with radial system. Axial system thus appeared as the easiest scaled-up system. Compared to lab scale, the conclusions observed at industrial scale were the same in terms of product quality, but different for robustness and flexibility, which confirmed the importance to test the systems at industrial scale before acquiring the equipment.

Issu de : 5th International granulation Workshop, 20-22 June 2011, Lausanne, SWITZERLAND; International audience; The objective of this study is to get a better understanding of dry powder coating process using experiments and numerical methods. Materials chosen as host particles are SUGLETS® (granules mainly consisting of sugar) and as invited particles, magnesium stearate (MgSt). These two materials are introduced into a high shear mixer (Cyclomix). Operations were performed at various mixing time and rotation speed of the Cyclomix. The surface morphology analysis has confirmed that Suglet particles are coated with MgSt. The product properties such as flowability, wettability and particle size distributions were also characterized. The particle motion in the Cyclomix has been simulated by using Discrete Element Method (DEM). Both number of collisions and the collision force frequency are calculated. The simulation shows an increase of the collision number with the rotational speeds. This result indicates that choosing higher rotation speeds should be better for the dry coating process as long as the particles are not broken down.

International audience; Experimental dry coating of guest particles on the surface of host particles is performed by mechanical forces in a high shear mixer called "Cyclomix". The studied system (a mixture of particles of sugar, "Suglet™" as host particles and magnesium stearate as guest particles) was chosen as a model one to achieve better understandings of the phenomena during mixing. To simulate the flow of host/guest particles in the mixer, the Discrete Element Method (DEM) was applied. Experimental results such as flowability and wettability can be explained by particles flows evolutions with different rotational speed or duration treatment inside the Cyclomix.

International audience; This review proposes an overview of the main trends explored by the pharmaceutical industry in order to develop a greener and smarter pharmacy minimizing any negative impact to the environment, and using more sustainable processes and drugs. If many drugs have their origin in nature, many active ingredients are ``toxic by design''. Some trials were carried out to design ``green pills'', or greener active ingredients ``benign by design'', avoiding the environmental pollution risk. More efforts were developed to reduce fine chemicals production footprints, and to decrease their E-factor. Biotechnology, and the use of enzymes for some transformation reactions is another good way of progress. Advanced galenics allows to reduce drug footprints on environment, since it can deliver the right dose of drug at the right time and at the right place, decrease the drug doses, and lower the wastes. Use of continuous processes is a strong trend in the area of primary and secondary pharmaceutical production. It is linked to the quality by design concept and to the in-process control by process analytical technology tools. Added to their diversification strategy, and despite their lower research productivity, and that more and more patented blockbusters become or will become generics soon, it will help the pharmaceutical companies to continue their development. At the same time, these efforts toward a greener pharmacy, and a social education of patients, will contribute to the health organization economies and to preserve the future of our planet.

National audience; Pas de résumé

Corpus callosum agenesis (CCA) is generally diagnosed in utero. Outcome appears to be better if the malformation is isolated. The aim of this study, which is the first one with a long (10 years) and standardized follow up, was to report cognitive abilities of children with isolated CCA diagnosed prenatally.We prospectively evaluated 17 children. Clinical examinations, neuropsychological tests were performed each year. School achievement and personal and familial data were collected.Twelve children completed the entire follow up. One child was finally considered to have associated CCA, because signs of fetal alcohol syndrome had become obvious. Of the 11 other children, three (27%) had borderline intelligence whereas the intelligence levels of eight (73%) were in the normal range, although half of these children experienced some difficulties in scholastic achievement. Neither epilepsy nor intellectual deficiency was noted and intellectual quotient scores correlated strongly with the mother's education level.Although prenatal diagnosis of isolated CCA is reliable, false postnatal diagnoses remain possible (10-20%) even with complete prenatal screening. Outcome is mostly favorable because intelligence is within the normal range for nearly 3/4 of the children. However, they frequently have mild learning difficulties.

International audience; An investigation into the effects of the operating conditions on a dry particle coating process has been performed with mixtures of sugar grains (Suglet®) and magnesium stearate (MgSt) as the host and invited particles, respectively. Dry coating has been carried out in a high shear mixer (Cyclomix®) at different processing times t, rotational speeds ω, and filling ratios J, of the mixer. In order to determine the effect of the operating conditions on the properties of the products, the changes in flowability and wettability of the mixtures as a function of ω, J and the operating time t have been analyzed. The flowability is found to be improved by longer processing times. Higher speeds of rotations can improve the flowability more rapidly. However, the flowability does not seem to be sensitive to the filling ratio and it may be concluded that operating with higher filling ratios gives higher production. A method is proposed to measure the mass fraction of invited particles on the surfaces of the host particles. This fraction is linked to the product properties as the flowability and wettability. This result can be predicted by a model using a coating rate constant kc which can be used to optimize dry coating processes to give desired properties of coated particles. This coating rate constant can define and represent the efficiency of the dry coating.

Issu de : 12th International Meeting on Radiation Processing (IMRP-12), AVIGNON, FRANCE, MAR 25-30, 2001; International audience; The methods (ESR, TL, GPC) developed to prove whether or not a foodstuff has been irradiated can be used to get the same proof in case of an irradiation treatment of drugs, excipients and cosmetic products.