Les pratiques culturales traditionnelles appauvrissent les sols en zone des savanes du Tchad

Les systèmes de culture des savanes tchadiennes sont à base de céréales et de coton. Le mode de gestion des terres est actuellement peu adapté à la conservation de la fertilité des sols. Par conséquent, les rendements des cultures sont stationnaires. L’étude a été menée dans les savanes au sud du Tchad et a eu pour objectif d’évaluer les effets des pratiques agricoles paysannes sur les rendements des cultures. Les données ont été complétées par la caractérisation physico-chimique des sols à partir des indices de déstructuration des sols qui ont été calculés. Quatre systèmes de culture (successions culturales) ont été identifiés : coton/sorgho, arachide/sorgho, sorgho/coton et maïs/arachide. Dans ces systèmes, les rendements des cultures et l’indice de déstructuration des sols ont baissé de 2006 à 2007. De même, les teneurs en carbone des sols du système maïs/arachide ont beaucoup diminué. Les sols étudiés se sont révélés particulièrement sensibles à l’érosion et pauvres en matière organique. Pour assurer une production agricole durable, nous préconisons l’emploi d’engrais organiques et minéraux ainsi que la culture des légumineuses.© 2010 International Formulae Group. All rights reserved. Mots clés : Céréales, arachide, pratiques culturales, fertilité du sol, système de culture

Optimized method for extraction of exosomes from human primary muscle cells

International audienceSkeletal muscle is increasingly considered an endocrine organ secreting myokines and extracellular vesicles (exosomes and microvesicles), which can affect physiological changes with an impact on different pathological conditions, including regenerative processes, aging, and myopathies. Primary human myoblasts are an essential tool to study the muscle vesicle secretome. Since their differentiation in conditioned media does not induce any signs of cell death or cell stress, artefactual effects from those processes are unlikely. However, adult human primary myoblasts senesce in long-term tissue culture, so a major technical challenge is posed by the need to avoid artefactual effects resulting from pre-senescent changes. Since these cells should be studied within a strictly controlled pre-senescent division count (30 times fewer differentiated myoblasts than what is required for the ultracentrifugation method. In addition, exosomes could still be integrated into recipient cells such as human myotubes or iPSC-derived motor neurons. Modified polymer-based precipitation combined with extra washing steps optimizes exosome yield from a lower number of differentiated myoblasts and less conditioned medium, avoiding senescence and allowing the execution of multiple experiments without exhausting the proliferative capacity of the myoblasts

Effets des fumures organo-minérales et des rotations culturales sur la qualité des sols

Les sols des régions semi-arides de l’Afrique de l’Ouest sont sujets à une dégradation graduelle de leur fertilité. Ils sont connus être d’une productivité très limitée et les moyens modernes pour augmenter et maintenir cette productivité sont le plus souvent très onéreux pour les petits exploitants. Ainsi, pour trouver de solutions probantes pour une gestion durable de la fertilité des sols, une étude a été conduite à la station de recherche agricole de Saria au Centre Ouest du Burkina Faso sur un dispositif implanté depuis 1960 où des fumures organique et/ou minérale combinées à des systèmes de cultures sont testés. L’objectif de cette étude est d’évaluer les effets de ces fumures combinés aux rotations culturales, sur le statut organique et la qualité des sols. Ce dispositif expérimental est du type Split splot à 6 répétitions, combinant six niveaux de fertilisation et à trois types de rotation. La monoculture de sorgho et la rotation sorgho/niébé ont été retenues dans cette étude. Des échantillons de sol ont été prélevés à 0-20 cm dans les parcelles du dispositif, et dans une jachère naturelle jouxtant le site expérimental et qui a servi de référence. La texture du sol et les teneurs en carbone et azote ont été déterminées. Les résultats obtenus montrent que, plus que le carbone organique total du sol, c’est plutôt sa fraction labile qui se révèle être un indicateur très sensible aux changements précoces qui affectent le statut organique des sols. En effet, l’indice de gestion du carbone qui apparait très corrélé avec la stabilité structurale du sol se positionne comme un outil fiable d’évaluation de la qualité des sols soumis aux pratiques culturales. Par ailleurs, les rotations sorgho-niébé se révèlent être des pratiques culturales les plus appropriées pour assurer une durabilité de la qualité des sols et des productions agricoles.© 2016 International Formulae Group. All rights reserved.Mots clés: Fumures minérales et organo-minérales, rotations culturales, carbone, azote, Burkina FasoEnglish Title: Effects of organo-mineral fertilizers and crop rotations on soil qualityEnglish AbstractIn the semiarid tropics of west Africa, agricultural lands are gradually degrading leading to decline soil fertitily. The low soil fertility is also increasingly recognized as a fundamental cause of declining food security in small-farm households. In this region, sustainable soil fertility management is constantly a challenge for crop productions. To come with an alternative soil fertility management practices, a trial was set up in 1960 in  the agricultural research station of Saria to identify the optimal soil management strategies for specific cropping systems to ensure sustainability of productivity. The experimental design is a split plot with 6 replications, with the main plots being three rotation system and the subplots consisting to 6 fertility management options. The main objective of this study was to evaluate the effects of soil fertility management practices combining crop rotation and fertility management options under continuous sorghum cropping and sorghum-cowpea (Vigna unguiculata) rotation systems in Burkina Faso on soil organic matter status and soil quality. Soil samples were collected at 0–20 cm depth. A bush fallow (more than 60 years old) of Andropogon gayanus located near the experimental siten is consider as natural condition for accessing changes due to landuse practices. Particle-sizes distribution, soil total N and carbon were determinated. The results indicated that labile organic pools are more sensitive to land use than total soil organic carbon. Carbon management index (CMI) which appeared highly correlated with soil structural stability is positioned as a reliable tool for soil quality assessment under several cropping systems. Furthermore, cowpea - sorghum rotation system seem to be the most appropriate cultural practice in ensuring sustainability of soil quality and agricultural production.© 2016 International Formulae Group. All rights reserved.Keywords: Mineral fertilizer combined with organic manure, crop rotation, carbon, nitrogen, Burkina Fas

Muscle cells of sporadic amyotrophic lateral sclerosis patients secrete neurotoxic vesicles

Background: The cause of the motor neuron (MN) death that drives terminal pathology in amyotrophic lateral sclerosis (ALS) remains unknown, and it is thought that the cellular environment of the MN may play a key role in MN survival. Several lines of evidence implicate vesicles in ALS, including that extracellular vesicles may carry toxic elements from astrocytes towards MNs, and that pathological proteins have been identified in circulating extracellular vesicles of sporadic ALS patients. Because MN degeneration at the neuromuscular junction is a feature of ALS, and muscle is a vesicle-secretory tissue, we hypothesized that muscle vesicles may be involved in ALS pathology. Methods: Sporadic ALS patients were confirmed to be ALS according to El Escorial criteria and were genotyped to test for classic gene mutations associated with ALS, and physical function was assessed using the ALSFRS-R score. Muscle biopsies of either mildly affected deltoids of ALS patients (n = 27) or deltoids of aged-matched healthy subjects (n = 30) were used for extraction of muscle stem cells, to perform immunohistology, or for electron microscopy. Muscle stem cells were characterized by immunostaining, RT-qPCR, and transcriptomic analysis. Secreted muscle vesicles were characterized by proteomic analysis, Western blot, NanoSight, and electron microscopy. The effects of muscle vesicles isolated from the culture medium of ALS and healthy myotubes were tested on healthy human-derived iPSC MNs and on healthy human myotubes, with untreated cells used as controls. Results: An accumulation of multivesicular bodies was observed in muscle biopsies of sporadic ALS patients by immunostaining and electron microscopy. Study of muscle biopsies and biopsy-derived denervation-naïve differentiated muscle stem cells (myotubes) revealed a consistent disease signature in ALS myotubes, including intracellular accumulation of exosome-like vesicles and disruption of RNA-processing. Compared with vesicles from healthy control myotubes, when administered to healthy MNs the vesicles of ALS myotubes induced shortened, less branched neurites, cell death, and disrupted localization of RNA and RNA-processing proteins. The RNA-processing protein FUS and a majority of its binding partners were present in ALS muscle vesicles, and toxicity was dependent on the expression level of FUS in recipient cells. Toxicity to recipient MNs was abolished by anti-CD63 immuno-blocking of vesicle uptake. Conclusions: ALS muscle vesicles are shown to be toxic to MNs, which establishes the skeletal muscle as a potential source of vesicle-mediated toxicity in ALS

Muscle cells of sporadic amyotrophic lateral sclerosis patients secrete neurotoxic vesicles

BACKGROUND: The cause of the motor neuron (MN) death that drives terminal pathology in amyotrophic lateral sclerosis (ALS) remains unknown, and it is thought that the cellular environment of the MN may play a key role in MN survival. Several lines of evidence implicate vesicles in ALS, including that extracellular vesicles may carry toxic elements from astrocytes towards MNs, and that pathological proteins have been identified in circulating extracellular vesicles of sporadic ALS patients. Because MN degeneration at the neuromuscular junction is a feature of ALS, and muscle is a vesicle-secretory tissue, we hypothesized that muscle vesicles may be involved in ALS pathology. METHODS: Sporadic ALS patients were confirmed to be ALS according to El Escorial criteria and were genotyped to test for classic gene mutations associated with ALS, and physical function was assessed using the ALSFRS-R score. Muscle biopsies of either mildly affected deltoids of ALS patients (n = 27) or deltoids of aged-matched healthy subjects (n = 30) were used for extraction of muscle stem cells, to perform immunohistology, or for electron microscopy. Muscle stem cells were characterized by immunostaining, RT-qPCR, and transcriptomic analysis. Secreted muscle vesicles were characterized by proteomic analysis, Western blot, NanoSight, and electron microscopy. The effects of muscle vesicles isolated from the culture medium of ALS and healthy myotubes were tested on healthy human-derived iPSC MNs and on healthy human myotubes, with untreated cells used as controls. RESULTS: An accumulation of multivesicular bodies was observed in muscle biopsies of sporadic ALS patients by immunostaining and electron microscopy. Study of muscle biopsies and biopsy-derived denervation-naïve differentiated muscle stem cells (myotubes) revealed a consistent disease signature in ALS myotubes, including intracellular accumulation of exosome-like vesicles and disruption of RNA-processing. Compared with vesicles from healthy control myotubes, when administered to healthy MNs the vesicles of ALS myotubes induced shortened, less branched neurites, cell death, and disrupted localization of RNA and RNA-processing proteins. The RNA-processing protein FUS and a majority of its binding partners were present in ALS muscle vesicles, and toxicity was dependent on the expression level of FUS in recipient cells. Toxicity to recipient MNs was abolished by anti-CD63 immuno-blocking of vesicle uptake. CONCLUSIONS: ALS muscle vesicles are shown to be toxic to MNs, which establishes the skeletal muscle as a potential source of vesicle-mediated toxicity in ALS

Congenital microcephaly: Case definition & guidelines for data collection, analysis, and presentation of safety data after maternal immunisation.

Need for developing case definitions and guidelines for data collection, analysis, and presentation for congenital microcephaly as an adverse event following maternal immunisation Congenital microcephaly, also referred to as primary microcephaly due to its presence in utero or at birth, is a descriptive term for a structural defect in which a fetus or infant’s head (cranium) circumference is smaller than expected when compared to other fetuses or infants of the same gestational age, sex and ethnic background

Identification of genes expressed in human cumulus cells according to oocyte nuclear maturity : effect of maturation conditions

Les cellules du cumulus (CCs) forment avec l'ovocyte le complexe ovocyte-cumulus (COC). Au cours de la folliculogénèse, un dialogue interdépendant régi par des jonctions communicantes se crée entre l'ovocyte et les CCs adjacentes. L'ovocyte en sécrétant certains facteurs, permettrait la différentiation et la prolifération des CCs qui, parallèlement, fournissent à l'ovocyte les nutriments nécessaires à sa maturation et son développement. La maturation nucléaire de l'ovocyte est définie par l'expulsion du 1er globule polaire au stade métaphase II (MII). Notre équipe a préalablement démontré que certains gènes exprimés dans les CCs chez l'humain, pouvaient prédire indirectement le potentiel implantatoire embryonnaire et la survenue d'une grossesse. Dans l'objectif d'identifier des marqueurs fiables de la maturité des ovocytes, nous avons analysé le profil transcriptomique des CCs en fonction du stade de maturité des ovocytes (VG, MI et MII). Dans un deuxième temps, nous avons étudié l'impact des conditions de maturation in vivo versus in vitro, sur le profil d'expression de gènes des CCs en fonction du stade de maturation ovocytaire. Nous avons mis en évidence, pour la première fois, une signature spécifique dans les CCs associée à la maturité nucléaire des ovocytes in vivo et in vitro. Nous avons également observé une sous-expression des gènes impliqués dans la maturation ovocytaire et l'expansion des CCs, et une sur-expression des gènes associés au cycle cellulaire dans les CCs dérivées d'ovocytes maturés in vitro, comparées aux CCs issus d'ovocytes in vivo. En comparant l'expression des gènes dans les CCs selon la condition de maturation et le stade de maturité de l'ovocyte, nous avons identifié deux voies de signalisation dominantes : la voie des lipides (transport du cholestérol et des triglycérides) fortement activée en condition in vivo, et le processus de réplication, recombinaison et réparation d'ADN, spécifique aux CCs in vitro. Nos résultats montrent que les conditions de maturation des COCs ont un impact sur la signature moléculaire des CCs. De plus, La signature moléculaire identifiée dans les CCs à différents stades de maturation ovocytaire nous a permis de définir la compétence des CCs. En considérant ce critère, nous avons observé que les ovocytes matures associés à des cellules du cumulus compétents (sur-expression de la signature des CCs au stade MII) présentent un potentiel de formation de blastocyste supérieur aux ovocytes MII entourés des cellules du cumulus incompétents (sur-expression de la signature des CCs au stade VG et/ou MI). Ces résultats ouvrent ainsi de nouvelles perspectives en application clinique. Des études supplémentaires sont toutefois nécessaires afin d'identifier, dans un premier temps, les facteurs qui influencent l'expression des gènes au cours de la maturation ovocytaire in vivo et comprendre ainsi les voies de signalisation altérées par les conditions de culture in vitro.Cumulus cells (CCs) associated with the oocyte form the cumulus-oocyte complex (COC). During folliculogenesis, interdependent dialogue governed by gap junctions is created between the oocyte and adjacent CCs. The oocyte, by secreting certain factors allows the differentiation and proliferation of CCs which, at the same time, provide nutrients to the oocyte for its maturation and development. Nuclear maturation of oocytes is defined by its transition from germinal vesicle (GV) to metaphase I (MI) up to metaphase II (MII) phase. Our team previously shown that certain genes expressed in the human CCs could predict embryo and the pregnancy outcomes. We analyzed the transcriptomic profile of CCs according to oocyte nuclear maturation stages (GV, MI and MII). The aim of this study was to identify the CCs molecular signature according to nuclear maturation oocyte under in vivo and in vitro conditions. In addition, we studied the impact of culture conditions of the COCs under in vivo and in vitro on the gene expression profile of CCs. We have demonstrated that there is a specific signature in the human CCs associated with the nuclear maturity of human oocytes whatever the culture condition. We have also observed the under-expression of genes involved in oocyte maturation and CCs expansion, and the over-expression of genes associated with cell cycle function in the CCS derived from in vitro versus in vivo oocytes. By comparing gene expression in the CCs according to oocyte nuclear maturation stages, we have identified two dominant signaling pathways: the lipids pathway (cholesterol transport and triglyceride) strongly activated in in vivo conditions, and the process of replication, recombination and DNA repair, which appear to be specific to in vitro CCs. Our results suggest that the maturation conditions of COCs have an impact on the molecular signature of CCs.Moreover, our data showed that matures oocytes can be surrounded by competent (sur-expression of the identified molecular signature of CCs derived from oocyte at MII stage) or incompetent CCs (sur-expression of the identified molecular signature of CCs derived from oocyte at GV or/and MI stages). These results open new perspectives in clinical application.Further studies are needed to identify factors influencing gene expression during oocyte maturation in vivo. These data should help to better understand how/why signaling pathways are altered by culture conditions in vitro