Molecular Identification of Broomrape Species from a Single Seed by High Resolution Melting Analysis

Broomrapes are holoparasitic plants spreading through seeds. Each plant produces hundreds of thousands of seeds which remain viable in the soils for decades. To limit their spread, drastic measures are being taken and the contamination of a commercial seed lot by a single broomrape seed can lead to its rejection. Considering that broomrapes species identification from a single seed is extremely difficult even for trained botanists and that among all the described species, only a few are really noxious for the crops, numerous seed lots are rejected because of the contamination by seeds of non-noxious broomrape species. The aim of this study was to develop and evaluate a High Resolution Melting assay identifying the eight most noxious and common broomrape species (P. aegyptiaca, O. cernua, O. crenata, O. cumana, O. foetida, O. hederae, O. minor, and P. ramosa) from a single seed. Based on trnL and rbcL plastidial genes amplification, the designed assay successfully identifies O. cumana, O. cernua, O. crenata, O. minor, O. hederae, and O. foetida; P. ramosa and P. aegyptiaca can be differentiated from other species but not from each other. Tested on 50 seed lots, obtained results perfectly matched identifications performed by sequencing. Through the analysis of common seed lots by different analysts, the reproducibility of the assay was evaluated at 90 %. Despite an original sample preparation process it was not possible to extract enough DNA from some seeds (10% of the samples). The described assay fulfils its objectives and allows an accurate identification of the targeted broomrape species. It can be used to identify contaminants in commercial seed lots or for any other purpose. The assay might be extended to vegetative material

Effects of sodium nitrite reduction, removal or replacement on cured and cooked meat for microbiological growth, food safety, colon ecosystem, and colorectal carcinogenesis in Fischer 344 rats

Epidemiological and experimental evidence indicated that processed meat consumption is associated with colorectal cancer risks. Several studies suggest the involvement of nitrite or nitrate additives via N-nitroso-compound formation (NOCs). Compared to the reference level (120 mg/kg of ham), sodium nitrite removal and reduction (90 mg/kg) similarly decreased preneoplastic lesions in F344 rats, but only reduction had an inhibitory effect on Listeria monocytogenes growth comparable to that obtained using the reference nitrite level and an effective lipid peroxidation control. Among the three nitrite salt alternatives tested, none of them led to a significant gain when compared to the reference level: vegetable stock, due to nitrate presence, was very similar to this reference nitrite level, yeast extract induced a strong luminal peroxidation and no decrease in preneoplastic lesions in rats despite the absence of NOCs, and polyphenol rich extract induced the clearest downward trend on preneoplastic lesions in rats but the concomitant presence of nitrosyl iron in feces. Except the vegetable stock, other alternatives were less efficient than sodium nitrite in reducing L. monocytogenes growth

Biomedicines

Antibody-mediated rejection (ABMR) is the leading cause of allograft failure in kidney transplantation. Its histological hallmark is represented by lesions of glomerulitis i.e., inflammatory cells within glomeruli. Current therapies for ABMR fail to prevent chronic allograft damage i.e., transplant glomerulopathy, leading to allograft loss. We used laser microdissection of glomeruli from formalin-fixed allograft biopsies combined with mass spectrometry-based proteomics to describe the proteome modification of 11 active and 10 chronic active ABMR cases compared to 8 stable graft controls. Of 1335 detected proteins, 77 were deregulated in glomerulitis compared to stable grafts, particularly involved in cellular stress mediated by interferons type I and II, leukocyte activation and microcirculation remodeling. Three proteins extracted from this protein profile, TYMP, WARS1 and GBP1, showed a consistent overexpression by immunohistochemistry in glomerular endothelial cells that may represent relevant markers of endothelial stress during active ABMR. In transplant glomerulopathy, 137 proteins were deregulated, which favor a complement-mediated mechanism, wound healing processes through coagulation activation and ultimately a remodeling of the glomerular extracellular matrix, as observed by light microscopy. This study brings novel information on glomerular proteomics of ABMR in kidney transplantation, and highlights potential targets of diagnostic and therapeutic interest

Model of gene correction of xeroderma pigmentosum mediated by engineered endonuclease-induced homologous recombination

Le xeroderma pigmentosum (XP) est une maladie génétique rare caractérisée par une hypersensibilité aux ultraviolets (UV) et une forte incidence des tumeurs cutanées. Les cellules des patients XP sont incapables d’éliminer les lésions induites dans l’ADN par les UV en raison d’un dysfonctionnement du mécanisme de réparation par excision de nucléotides (NER). Plusieurs groupes de complémentation ont été identifiés dans le syndrome XP, parmi lesquels le groupe XP-C représente la majorité des patients à travers le monde.Au cours de mon travail de thèse, j’ai développé un modèle de correction ciblée par recombinaison homologue (RH) d’une délétion de deux nucléotides au niveau de l’exon 9 du gène XPC aboutissant à l’apparition prématurée d’un codon stop. Afin de stimuler la RH, deux types de nucléases ingéniérées sont utilisées : les méganucléases et les TALENs. J’ai observé que la méthylation de la séquence ciblée pouvait affecter l’activité de celles-ci et donc l’efficacité du ciblage de gène. Cependant, deux approches ont été développées pour résoudre ce problème : l’utilisation d’un agent déméthylant (5-aza-2’-désoxycytidine (5azadC)) ou la création d’une endonucléase insensible à la méthylation. L’utilisation des méganucléases en combinaison avec la 5azadC a permis de stimuler la fréquence de coupure de presque 20 fois dans des fibroblastes XPC et la TALEN modifiée permet une augmentation de 40 fois. Avec ces deux stratégies j’ai obtenu des événements de correction génétique par introduction d’une matrice de réparation dans le locus ciblé avec une fréquence proche de 3%. La caractérisation des clones corrigés avec la TALEN XPC montre la correction génomique des deux nucléotides dans l’exon 9, une restauration de l’expression de la protéine XPC et une résistance cellulaire après irradiation UV traduisant le rétablissement des fonctions de la NER. Cette étude représente la première preuve de correction génétique de cellules déficientes en protéine XPC en utilisant une approche ciblée.Xeroderma pigmentosum (XP) is a rare inherited genetic disorder characterized by an UV hypersensitivity and a severe predisposition to skin cancers. Cells from XP patients are deficient in nucleotide excision repair (NER) of UV‐induced DNA lesions. Several complementation groups have been identified in the XP syndrome and the XP-C group represents the majority of XP patients around the world. During my PhD work, I developed a model of targeted correction by homologous recombination (HR) in order to correct a deletion of two nucleotides in the ninth exon in XPC gene leading to a premature stop codon. To stimulate HR, I used two types of engineered endonucleases : meganucleases and TALEN. I observed that the target methylation status could affect the endonuclease activities and therefore XPC gene correction. Nervertheless, I developed two approaches to overcome this methylation sensitivity : use of a demethylating agent (5-aza-2-deoxycytidine (5azadC)) or a specific engineering of TALEN. Using 5azadC with meganuclease allowed to stimulate the cutting frequency by nearly 20 fold in XPC fibroblasts and the engineered TALEN allowed a 40 fold-increase in frequency. With both strategies I obtained genetic correction events by repair matrix introduction in the targeted locus with a near 3% frequency. The characterization of corrected clones with the XPC TALEN shows genomic correction in the ninth exon, a restoration of the XPC protein expression and cell survival following UV exposure, thus demonstrating fully recovered normal repair activity by NER. This study represents the first evidence of genetic correction of XPC-deficient cells by a targeted approach

Stabilisation de l'interface liquide-liquide dans un contacteur membranaire : Application à l'extraction sélective de terpènes oxygénés d'huile essentielle d'agrumes

Citrus essential oils are an abundant and inexpensive source of oxygenated terpenes, which are highly valuable molecules in particular for the flavor and fragrance industry. Their industrial liquid-liquid extraction can potentially be intensified by the use of membrane contactors which eliminate the need for a decantation step and afford a broader choice of solvents. However, the low interfacial tension between the citrus oils and the hydro-alcoholic solvents typically used for extraction makes interface stabilization very challenging. In fact, it requires such a tight process control that the use of membrane contactors is currently not feasible on industrial scale. The aim of this work was therefore to propose solutions to facilitate the interface stabilization in membrane contactors. Practically, this consists in the broadening of the pressure difference range between the two phases must be controlled. A pressure difference of 0.5 bar was selected as a minimum to apply this technology at industrial scale. This means that the breakthrough pressure ΔPc, i.e. the pressure difference above which the non wetting phase passes through the membrane, has to be above 0.5 bar. Two approaches were investigated: firstly, to select the best suited solvent, and secondly to identify membranes with small maximum diameter of pores and appropriate surface properties. In conclusion, this study afforded the identification of membranes in PTFE and grafted ceramic for which the pressure difference between the two phases can be controlled in a relatively broad window of 0.5 bar. It represents a significant advance compared to commercially available Liqui-Cel® modules in polypropylene.Les huiles essentielles d'agrumes sont une source abondante et peu chère de terpènes oxygénés, molécules à haute valeur ajoutée pour l'industrie des arômes et des parfums notamment. Le procédé industriel employé pour leur extraction, de type liquide-liquide peut potentiellement être intensifié par l'utilisation de contacteurs membranaires, qui permettent de s'affranchir de l'étape de décantation et d'élargir le choix des solvants. Cependant, la faible tension de surface entre les huiles essentielles et les solvants hydro-alcooliques rend difficile la stabilisation de l'interface dans la membrane. En l'état actuel, les contacteurs membranaires sont inenvisageables pour cette application à l'échelle industrielle. L'objectif est de proposer des solutions afin de faciliter la stabilisation de l'interface. On souhaite élargir la fenêtre dans laquelle la différence de pression entre les deux phases doit être contrôlée, une différence de 0,5 bar étant choisie comme minimum pour l'application à plus grande échelle. Cela signifie que la pression critique ΔPc (au-dessus de laquelle la phase non mouillante traverse la membrane) doit être supérieure à 0,5 bar. Deux pistes ont été explorées : travailler d'une part sur le choix du solvant, et d'autre part sur la sélection de la membrane en fonction du diamètre maximal des pores ainsi que de ses propriétés de surface. En conclusion, cette étude a permis d'identifier des membranes en PTFE et céramique greffée pour lesquelles la différence de pression entre les deux phases peut être contrôlée dans une fenêtre relativement large de 0,5 bar. C'est une avancée considérable par rapport aux modules commerciaux Liqui-Cel® en polypropylène

Représentations et usages du parfum

Associés à la vie comme à la mort, à la beauté et au bien-être, aux soins du corps et aux plaisirs charnels, les parfums ont, de tout temps, occupé une place prépondérante dans la vie des hommes. Afin d’appréhender le parfum, évanescent, mais omniprésent dans notre société, il faut faire appel à ses origines, à ses représentations et à ses résonances physiologiques, culturelles et sociales. Le parfum demeure, en effet, porteur de significations et pénètre dans les sphères du sensoriel, des pratiques corporelles et du symbolique. Devenu produit industriel, le parfum dépasse le cadre de l’alchimie des senteurs. Cette recherche propose de mettre en perspective une possible dimension sociologique du parfum lorsqu’il est appréhendé dans sa globalité : représentations et usages, communication, conditionnement et publicité.Having linked to life as to death, to beauty and to well-being, to care of the body and to carnal pleasures, perfumes, any time long, occupied a preponderant place in the life of men. To studied perfume, evanescent, but omnipresent in our society, it is necessary to appeal to its origins, its representations and its physiological, cultural and social resonance. Perfume remains, indeed, bearing of significations and enters the spheres of the sensory, of bodily practices and of symbolism. Become industrial product, perfume exceeds the frame of alchemy fragrances. This research offers to put into relief a possible sociological dimension of perfume studied taken as a whole: representations and uses, communication, packaging and advertising

Membrane-based oil-water separation: Membranes, concepts and case studies

1. IntroductionThe demand for efficient oil-water separation is driven by the tightening of discharge limits for e.g. produced water in the oil and gas industry, process water in the petrochemical industry and bilge water in the marine industry. The global produced water production alone is over 71 billion bbl/a – 8.4 billion m3/a [1]. Among the different technologies, particularly ultrafiltration with polymeric membranes can be used to reduce oil in water down to less than 1 ppm leading to an energy-efficient and compact approach for the industry. The first part of the presentation will introduce the ETNA membrane for oil-water separation. The second part will review different oil-water separation technologies with focus on ultrafiltration-based synergy processes, while the final part will present two case studies on industrial oil-water separation processes. 2. Oil-water separation membranesThe membranes chosen for oil-water separation are the ETNA01PP and ETNA10PP membranes (Alfa Laval Nakskov A/S, Denmark) with a molecular weight cut-off of 1,000 and 10,000 Dalton, respectively. Both membranes are surface-modified PVDF membranes on a polypropylene support and are permanently hydrophilic. The hydrophilic surface of these membranes increases the retention of hydrophobic compounds such as oil and thus reduces membrane fouling [2].3. Membrane-based oil-water separation conceptsThe most common technologies in the industry for oil-water separation are skimmers (gravity/density), coalescers (coalescence) and centrifugal separators (centrifugation/density). Skimmers, centrifugal separators and coalescers are very efficient separation processes for initial oil-water separation but their final oil-in-water levels are commonly higher than the level of less than 1 ppm which is achievable by ultrafiltration. Since the industry often requires high oil recovery combined with low oil content in the discharge water, the optimal solution is often a process combination consisting of a conventional separation technology followed by ultrafiltration and thus different ultrafiltration-based oil-water separation concepts will be reviewed.4. Case studies: Industrial scale oil-water separation The potential of ultrafiltration-based concepts will be highlighted in two case studies. The first plant has a capacity of 40,000 tons/a treating bilge water and industrial oil waste with low suspended solids concentration. The process set-up consists of a skimmer, high speed centrifugal separators and paper band filter plus ultrafiltration. The second plant has a capacity of 80,000 tons/a handling oily waste from ships, petrochemical facilities or offshore industry with higher levels of suspended solids. In the process set-up, decanter centrifuges, high speed separators and evaporation are used in combination with ultrafiltration. Both process set-ups result in an oil-rich phase with more than 98% oil and a water rich-phase with less than 2 ppm oil. 5. Conclusion and outlookOverall, this paper shows that ultrafiltration-based oil-water separation concepts can achieve both high oil concentrations combined with very low residual oil in the water phase and offer therefore an excellent alternative for current and future oil-water separation challenges

Meeting the challenges: Membrane processes for water recovery from oily and PVC wastewater

The demand for water recovery in the industry is increasingly important partly due to water shortage and partly due to tightening of regulatory requirements. In this paper water recovery solutions for two challenging types of wastewater - oily and polyvinylchloride (PVC) wastewater - are presented. The first part of the presentation analyses the operation and performance of a membrane unit operated in an oily waste treatment facility. The facility handles mainly bilge water and industrial oil waste with low concentration of suspended solids. The waste is pre-treated in skimmer tank followed by a band filter before entering an ultrafiltration unit with polymeric membranes. At the entrance of the ultrafiltration unit the oily wastewater contains approx. 1000 ppm emulsified oil, which is then separated into an oil-rich stream which is recycled to the skimmer tank and a permeate stream with less than 2 ppm oil sufficient for direct discharge. The second part of the presentation focuses on the recovery and reuse of water in the polyvinylchloride (PVC) production. In the PVC production approx. 2.0 - 2.5 m3 of demineralised water is required per ton PVC. In today’s installations 20% of the water used is lost during drying of the PVC or as sealing water. The remaining water is recovered by the PVC decanter. 20 – 25% of the water recovered by the PVC decanter is used for flushing of the facilities, while 75 – 80% is currently discharged to biological treatment. A new concept based on reverse osmosis does not only remove the residual PVC particles but also the inhibitors and conductivity down to levels allowing the direct recycle of decanter water to the polymerisation step and thus reducing water consumption and water treatment costs – both in-take and discharge cost - significantly. Overall, the case studies presented will demonstrate how membranes can handle even challenging industrial wastewaters at minimal energy costs

Développement d'un modèle de correction génétique du xeroderma pigmentosum par recombinaison homologue ciblée par des endonucléases ingéniérées.

Le xeroderma pigmentosum (XP) est une maladie génétique rare caractérisée par une hypersensibilité aux ultraviolets (UV) et une forte incidence des tumeurs cutanées. Les cellules des patients XP sont incapables d éliminer les lésions induites dans l ADN par les UV en raison d un dysfonctionnement du mécanisme de réparation par excision de nucléotides (NER). Plusieurs groupes de complémentation ont été identifiés dans le syndrome XP, parmi lesquels le groupe XP-C représente la majorité des patients à travers le monde.Au cours de mon travail de thèse, j ai développé un modèle de correction ciblée par recombinaison homologue (RH) d une délétion de deux nucléotides au niveau de l exon 9 du gène XPC aboutissant à l apparition prématurée d un codon stop. Afin de stimuler la RH, deux types de nucléases ingéniérées sont utilisées : les méganucléases et les TALENs. J ai observé que la méthylation de la séquence ciblée pouvait affecter l activité de celles-ci et donc l efficacité du ciblage de gène. Cependant, deux approches ont été développées pour résoudre ce problème : l utilisation d un agent déméthylant (5-aza-2 -désoxycytidine (5azadC)) ou la création d une endonucléase insensible à la méthylation. L utilisation des méganucléases en combinaison avec la 5azadC a permis de stimuler la fréquence de coupure de presque 20 fois dans des fibroblastes XPC et la TALEN modifiée permet une augmentation de 40 fois. Avec ces deux stratégies j ai obtenu des événements de correction génétique par introduction d une matrice de réparation dans le locus ciblé avec une fréquence proche de 3%. La caractérisation des clones corrigés avec la TALEN XPC montre la correction génomique des deux nucléotides dans l exon 9, une restauration de l expression de la protéine XPC et une résistance cellulaire après irradiation UV traduisant le rétablissement des fonctions de la NER. Cette étude représente la première preuve de correction génétique de cellules déficientes en protéine XPC en utilisant une approche ciblée.Xeroderma pigmentosum (XP) is a rare inherited genetic disorder characterized by an UV hypersensitivity and a severe predisposition to skin cancers. Cells from XP patients are deficient in nucleotide excision repair (NER) of UV induced DNA lesions. Several complementation groups have been identified in the XP syndrome and the XP-C group represents the majority of XP patients around the world. During my PhD work, I developed a model of targeted correction by homologous recombination (HR) in order to correct a deletion of two nucleotides in the ninth exon in XPC gene leading to a premature stop codon. To stimulate HR, I used two types of engineered endonucleases : meganucleases and TALEN. I observed that the target methylation status could affect the endonuclease activities and therefore XPC gene correction. Nervertheless, I developed two approaches to overcome this methylation sensitivity : use of a demethylating agent (5-aza-2-deoxycytidine (5azadC)) or a specific engineering of TALEN. Using 5azadC with meganuclease allowed to stimulate the cutting frequency by nearly 20 fold in XPC fibroblasts and the engineered TALEN allowed a 40 fold-increase in frequency. With both strategies I obtained genetic correction events by repair matrix introduction in the targeted locus with a near 3% frequency. The characterization of corrected clones with the XPC TALEN shows genomic correction in the ninth exon, a restoration of the XPC protein expression and cell survival following UV exposure, thus demonstrating fully recovered normal repair activity by NER. This study represents the first evidence of genetic correction of XPC-deficient cells by a targeted approach.PARIS11-SCD-Bib. électronique (914719901) / SudocSudocFranceF