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Abuse of tax law is both a complex and ambiguous concept per se. Added to the fact that each country has its own approach, which entails differing consequences as regards domestic, European community and international tax law, abuse of tax law across Europe may become a real puzzle. A thorough analysis of such concept requires both to bring out common features, shared by all domestic approaches of the countries at stake – i.e., France, Germany, Italy, Spain, and the UK – but also to describe the theoretical and practical specificities of each approach. The second part of this article, which apprehends the Italian, Spanish and British approaches, completes the first scan of the similarities and dissimilarities of the same concept, viewed through varied domestic prisms (for the French and German approaches, see EC Tax Review 2010-2). For example, on the one hand, Italy, Spain and the UK all have a prior ruling procedure, and apply no specific penalty to abusive schemes – as France does for example. On the other hand however, their approaches differ on a certain number of topics: the obligation of disclosure, for example, is compulsory in some cases in the UK whereas there is no such obligation in Italy or in Spain.

Abuse of tax law is both a complex and an ambiguous concept per se. Each country having developed its own approach, which entails differing consequences as regards domestic, EU and international tax law, abuse of tax law across Europe is a real puzzle. A thorough analysis of this concept requires both to bring out the common features shared by all domestic approaches of the countries chosen for this analysis – that is, France, Germany, Italy, Spain, and the United Kingdom – but also to describe the theoretical and practical specificities of each domestic approach. The first part of this article, which details the French and the German approaches, gives a taste of the similarities and of the differences of interpretation and practical implementation of the abuse of law concept viewed through two domestic prisms. For example, on the one hand, France and Germany both use a general concept of fraud, together with a certain number of specific anti-avoidance rules, and they both share a mostly statutory approach of the notion. On the other hand, however, the French and German approaches greatly differ on the penalty as well as on the proceedings which are applicable, whenever an abuse of law is characterized.

In a decision rendered on 27 November 2008 (C-418/07, Sté Papillon v. Ministère du Budget), the European Court of Justice ruled that the impossibility – resulting from the rules set forth in the French Tax Code – for a French parent company to include within a French tax consolidation group a French sub-subsidiary held through a subsidiary located in another Member State constitutes a restriction on the freedom of establishment. This decision raises several questions as to the application of domestic tax consolidation rules (as currently applicable in France and in other Member States) in cross-border situations and might be a first step toward a new era in European tax treatment of groups.

Stress plays an important role in the development of symptoms contributing to disease. Stress induces various disorders with gastrointestinal, physical, and psychological symptoms. Probiotics can help regulate or modulate gastrointestinal functions. The aim of the present study was to investigate the effects of a probiotic preparation (Probio-Stick) on stress-induced symptoms in volunteers. A double-blind, placebo-controlled, randomized study was conducted on volunteers with symptoms of stress. Subjects received a probiotic (Probio-Stick; Lallemand SAS, Saint-Simon, France) containing Lactobacillus acidophilus Rosell-52 and Bifidobacterium longum Rosell-175 (3 x 10(9) colony-forming units per sachet stick) or a sensorially identical placebo without probiotics during a 3-week period. The consumption of probiotics significantly reduced 2 stress-induced gastrointestinal symptoms (abdominal pain and nausea/vomiting) for intention-to-treat or per-protocol populations. In contrast, the probiotics did not significantly modify the other physical and psychological symptoms and sleep problems induced by stressful life events for intention-to-treat or per-protocol populations. The results indicate that Probio-Stick can provide a beneficial effect on the gastrointestinal symptoms experienced by individuals affected by chronic stress.

This three-part article discusses the compatibility of exit tax legislation applicable to corporate taxpayers in France, Germany, Italy, the Netherlands, Portugal, Spain and the United Kingdom with the EU freedom of establishment, especially in the light of the ECJ’s landmark National Grid decision. Part 1 scrutinizes whether a company transferring its tax residence or effecting an outbound cross-border conversion has access to Articles 49 and 54 TFEU under the laws of the Member State. It also addresses whether these laws restrict Article 49, and, if so, whether the restriction can be justified and is appropriate to ensure the attainment of its objective. Parts 2 and 3, each to be published in subsequent Intertax issues, review whether the legislation is proportional, address which other transactions the ECJ’s exit tax principles apply to, and provide conclusions and recommendations.

Live Saccharomyces cerevisiae yeast strains are widely used as feed additives for dairy cow s. Although, perfectly safe and applied in several food processes, the potential contamination of milk by the strain and more generally its possible influence on the microbial profile of milk could be questioned, this concerns particularly traditi onal cheeses made with raw milk. In an attempt to address these questions, two experiments were performed: A trial in an experimental farm , where two groups of cows, one receiving yeast supplementation and the other one being an unsupplemented control, were compared and a field trial where herds received alternatively yeast-supplemented and control diets. In both cases, viable counts of different microbial groups, including yeasts and molds in general and S. cerevisiae in particular were analysed in tank milk and results from suppleme nted groups or periods were compared with control ones. No v iable S. cerevisiae were detected in any of the samples analysed. Moreover, no n oticeable differences in the microbial profiles were observed. This applies also to the evolution of the microbial composition during an experimental cheese processing trial. These data sug gest that live yeast additive distributed to dairy cows does not influence milk microbial composition and cheese-making abilities.

Six years of new acquisitions at the library of the Institute for Higher Chinese Studies (Institut des Hautes Études Chinoises) (2001-2006). In the last 15 years, Chinese publishers have produced a growing number of collections, both small and large, bringing together books and documents from the Empire, of which the four largest are taken from the Library of the Four Treasuries compiled at the order of the Qianlong emperor. The Institute for Higher Chinese Studies (Institut des Hautes Études Chinoises) of the Collège de France strives to collect the best out of all these new publications. Shum Wing-Fong and Pierre-Henri Durand have previously written a survey of the publications acquired by the Institute from 1996 to 2000, and they now offer an overview of acquisitions since that time, presenting works whose diversity and importance deserve everyone’s attention., Depuis une quinzaine d’années, l’édition chinoise a multiplié la publication de collections, grandes ou petites, rassemblant livres et documents du temps de l'empire, dont les quatre plus monumentales se réclament de la fameuse Bibliothèque des quatre trésors compilée par la volonté de l’empereur Qianlong. L'Institut des Hautes Études Chinoises du Collège de France s'efforce de récolter au mieux cette manne providentielle. Shum Wing-Fong et Pierre-Henri Durand en avaient jadis donné un aperçu pour les années 1996 à 2000, ils nous proposent aujourd'hui une vue des principales acquisitions faites depuis lors, présentant des ouvrages dont la diversité et l'importance méritent de retenir l'attention générale., Shen Yongfang, Durand Pierre-henri. Six ans d’acquisitions nouvelles à la bibliothèque de l’Institut des Hautes Études Chinoises (2001-2006). In: Études chinoises, n°26, 2007. pp. 129-162.

Born in Huizhou in 1650, Zhang Chao was one of the leading citizens of the prosperous city of Yangzhou, where he became wealthy in the salt trade. An accomplished man of letters always ready to assist his confreres, many of whose books he published, he himself wrote a famous collection of aphorisms and witticisms, Shadows of Buried Dreams, endorsed by 700 comments from 115 of his friends. A careful examination of these comments, with the help of the author's own correspondence, lets us understand the genesis of this work, whose original plates gave rise to several succeeding editions, each augmented by the latest comments received. Shadows of Dreams illustrates a notable feature of Chinese xylography, better fitted to follow changes in the text over time than our Western printing, dedicated to producing editions on an ad hoc basis. Zhang Chao's book, which is seeing a revival of interest in today's China, is not simply a literary work. It is a social document revealing the network of friendships patiently woven by a wise man of the world., Né en 1650 à Huizhou, Zhang Chao fut l'une des figures de la prospère cité de Yangzhou, où il s'enrichit dans le commerce du sel. Homme de lettres accompli, toujours prêt à aider ses pairs, dont il édita bien des livres, il écrivit un célèbre recueil d'aphorismes et de bons mots, les Ombres de rêves enfouis, que rehaussent sept cents apostilles offertes par cent quinze de ses amis. Leur examen attentif permet, avec le secours de la correspondance de l'auteur, d'appréhender la genèse d'un ouvrage dont les planches originales autorisèrent plusieurs éditions successives, chacune augmentée des dernières apostilles reçues. Les Ombres de rêves illustrent un trait remarquable de la xylographie chinoise, mieux armée pour suivre les variations d'un texte dans le temps que notre imprimerie d'Occident, vouée à produire ses éditions au coup par coup. Le livre de Zhang Chao, qui connaît un regain de succès dans la Chine d'aujourd'hui, n'est pas seulement une œuvre littéraire, il est aussi un ouvrage mondain révélant le réseau d'amitiés patiemment tissé par un homme fort avisé., Durand Pierre-Henri. Zhang Chao et les Ombres de Rêves ou les amitiés d'un éditeur au temps de l'empereur Kangxi. In: Bulletin de l'Ecole française d'Extrême-Orient. Tome 95-96, 2008. pp. 363-405.

International audience; Background and aims: Intestinal barrier impairment is incriminated in the pathophysiology of intestinal gut disorders associated with psychiatric comorbidity. Increased intestinal permeability associated with upload of lipopolysaccharides (LPS) translocation induces depressive symptoms. Gut microbiota and probiotics alter behavior and brain neurochemistry. Since Lactobacillus farciminis suppresses stress-induced hyperpermeability, we examined whether (i) L. farciminis affects the HPA axis stress response, (ii) stress induces changes in LPS translocation and central cytokine expression which may be reversed by L. farciminis, (iii) the prevention of "leaky" gut and LPS upload are involved in these effects. Methods: At the end of the following treatments female rats were submitted to a partial restraint stress (PRS) or sham-PRS: (i) oral administration of L. farciminis during 2 weeks, (ii) intraperitoneat administration of ML-7 (a specific myosin light chain kinase inhibitor), (iii) antibiotic administration in drinking water during 12 days. After PRS or sham-PRS session, we evaluated LPS levels in portal blood, plasma corticosterone and adrenocorticotropic hormone (ACTH) levels, hypothalamic corticotropin releasing factor (CRF) and pro-inflammatory cytokine mRNA expression, and colonic paracellular permeability (CPP). Results: PRS increased plasma ACTH and corticosterone; hypothalamic CRF and pro-inflammatory cytokine expression; CPP and portal blood concentration of LPS. L. farciminis and ML-7 suppressed stress-induced hyperpermeability, endotoxemia and prevented HPA axis stress response and neuroinflammation. Antibiotic reduction of luminal LPS concentration prevented HPA axis stress response and increased hypothalamic expression of pro-inflammatory cytokines. Conclusion: The attenuation of the HPA axis response to stress by L. farciminis depends upon the prevention of intestinal barrier impairment and decrease of circulating LPS levels. (c) 2012 Elsevier Ltd. All rights reserved.

Pierre-Henri Durand : Bureaucratie Language and History. A few Consi¬ derations on the Grand Council and Macartney's Embassy to China. When Lord Macartney and his companions arrived at the Court of Emperor Qianlong in 1793, the Grand Council had reached its apogee. After half a century of existence, it had become so essential that, according to Beatrice Bartlett in her book Monarchs and Ministers , the ruler had fallen into the dependence of his close ministers : the autocratic empire had turned into a monarchy of councils. The British embassy gave rise to the circulation of a mass of documents between the high officials who were in charge of the delegation and the throne, that is the emperor and the Grand Council. By scrutinizing the words and the forms being used, this article brings to light a sort of administrative diplomacy and reveals the low degree of information provided by a language constrained by repetitive and stereo¬ typed conventions. These English testimonies throw light on the clichés, silences, and dark corners of the Chinese archives, while showing the limits of the official documentation so as to make tangible the complexity of historical reality and the workings of the political backstage. Philip Kuhn's brilliant book Soulstealers on the great 1768 sorcery case illustrates the difficulty for the historian to come to grips with the actual men and actors behind the representations of bureaucratic language. However, his picture of bureaucratic monarchy that left ample room to imperial arbitrary power seems hardly compatible with Beatrice Bartlett's ministerial monarchy., Lorsque Lord Macartney et ses compagnons arrivèrent à la cour de Qianlong en 1793, le Grand Conseil était dans son âge d'or. Après plus d'un demi-siècle d'existence, il était devenu si essentiel que, selon le Monarchs and Ministers de Beatrice Bartlett, le souverain était tombé dans la dépendance de ses proches ministres : l'empire autocratique s 'était mué en une monarchie des conseillers. Les Britanniques suscitèrent un flot de documents entre les hauts fonctionnaires chargés de les conduire et le Trône — l'empereur et le Grand Conseil. En prêtant attention au mot et à la forme, l'article met en évidence une diplomatique administrative et souligne le faible rende¬ ment informatif d'un langage pris dans l'étau des répétitions, des conven¬ tions et des stéréotypes. Les témoignages des Anglais éclairent les clichés, occultations et silences des archives chinoises ; il révèlent les limites de la matière officielle pour appréhender la complexité de la réalité histori¬ que et les dessous du jeu politique. Le brillant Soulstealers de Philip Kuhn sur le grand procès de sorcellerie de 1 768 illustre la difficulté pour 1 ' historien de saisir les hommes et les acteurs derrière les représentations du langage bureaucratique. Son modèle d'une monarchie bureaucratique laissant une large part à l'arbitraire impérial semble peu compatible avec la monarchie ministérielle de Beatrice Bartlett., Durand Pierre-henri. Langage bureaucratique et histoire : Variations autour du Grand Conseil et de l'ambassade Macartney. In: Études chinoises, vol. 12, n°1, Printemps 1993. pp. 41-145.

Background and aims: The purpose of this study was to investigate the efficacy of a synbiotic supplementation in reducing common winter diseases in children. Methods: A randomized, double-blind, placebo-controlled, multicentre study was conducted in young school-age children (3—7 years old) during a winter period. Participants were otherwise healthy children who suffered from at least three episodes of ear, nose and throat (ENT), respiratory tract or gastrointestinal illness during the previous winter. They were supplemented daily with either a synbiotic preparation (Lactobacillus helveticus R0052, Bifidobacterium infantis R0033, Bifidobacterium bifidum R0071, and fructooligosaccharide) or a matched placebo for 3 months. Over this period, all emergent health episodes of any type were recorded by parents in a diary. They were checked by investigators at regular monthly visits. The main study outcome was the percentage of children free of any episode during the study course. Results: We randomized 135 children (mean age: 4.1±1.0 years) to the synbiotic group ( n = 62) or placebo ( n = 73) group. At least one illness episode was reported in 32 children in the synbiotic group and 50 in the placebo group (51.6% versus 68.5%). This corresponded to a significant 25% relative risk reduction (95% CI 0.6—44.3%; p = 0.045). This difference was due to a decrease in the number of children who suffered from at least one ENT, respiratory tract or gastrointestinal disorder (50.0% with synbiotic versus 67.1% with placebo; p = 0.044). At least one sickness school day loss was noted in 25.8% of children with the synbiotic as compared with 42.5% with placebo ( p = 0.043). No treatment related side effects were detected in either group. Conclusions: This study suggests that a 3-month supplementation with this synbiotic preparation can decrease the risk of occurrence of common infectious diseases in children and limits the risk of school day loss.

International audience; Probiotics and prebiotics have been demonstrated to positively modulate the intestinal microflora and could promote host health. Although some studies have been performed on combinations of probiotics and prebiotics, constituting synbiotics, results on the synergistic effects tend to be discordant in the published works. The first aim of our study was to screen some lactic acid bacteria on the basis of probiotic characteristics (resistance to intestinal conditions, inhibition of pathogenic strains). Bifidobacterium was the most resistant genus whereas Lactobacillus farciminis was strongly inhibited. The inhibitory effect on pathogen growth was strain dependent but lactobacilli were the most effective, especially L farciminis. The second aim of the work was to select glucooligosaccharides for their ability to support the growth of the probiotics tested. We demonstrated the selective fermentability of oligodextran and oligoalternan by probiotic bacteria, especially the bifidobacteria, for shorter degrees of polymerisation and absence of metabolism by pathogenic bacteria. Thus, the observed characteristics confer potential prebiotic properties on these glucooligosaccharides, to be further confirmed in vivo, and suggest some possible applications in synbiotic combinations with the selected probiotics. Furthermore, the distinctive patterns of the different genera suggest a combination of lactobacilli and bifidobacteria with complementary probiotic effects in addition to the prebiotic ones. These associations should be further evaluated for their synbiotic effects through in vitro and in vivo models. (C) 2010 Elsevier Ltd. All rights reserved.

A convenient and efficient transformation system has been developed for the filamentous fungus Tolypocladium geodes. In contrast to most of the commonly described techniques requiring prior preparation of protoplasts or spheroplasts, this method leads to high efficiency transformation of T. geodes conidiospores following moderate lytic enzyme treatment. Competent cells so obtained are still resistant to osmotic pressure and can be stored frozen without loss of viability. The highest transformation frequency (3-5 x 10(3) transformants per microgram of DNA) was obtained with plasmid pUT737 containing the Sh ble gene conferring phleomycin resistance under the control of a strong promoter isolated from Trichoderma reesei. Southern hybridization revealed multiple integration sites of plasmid DNA into the T. geodes nuclear DNA despite the absence of homology between the transforming DNA and the recipient genome. Instability could not be detected for the phleomycin phenotype during more than five generations of mitotic growth under non-selective conditions.

A new Penicillium strain characterized by high yields of extracellular cellulase production, particularly β-glucosidase, was isolated from soil. On the basis of growth characteristics, reverse coloration, and structure of Penicilli, this strain was assigned to a new species, named Penicillium occitanis. The organism was subjected to a strain-improvement program, and a mutant (Pol6) was selected after several rounds of mutagenesis with NTG, EMS, and UV. Selection was for strains that were resistant to catabolite repression and were enhanced in cellulase and pectinase production. The strain Pol6 was also found to produce other polysaccharide-hydrolysing enzymes when grown on various inducing substrates. The production of polysaccharidases by Penicillium occitanis strain Pol6, under various fermentation conditions, and their preliminary separation by Fast Protein Liquid Chromatography is discussed.

L'hydrolyse de solutions de monofluorophosphates a ete etudiee par chromatographie ionique. Nous avons montre que le facteur predominant est le pH. En effet la constante de vitesse est quarante fois plus elevee a pH = 2 qu'a pH = 4, quelle que soit la temperature. L'hydrolyse est peu sensible a la nature du cation present dans la solution et a celle de l'acide rajoute pour obtenir le pH desire. Cette reaction d'hydrolyse du premier ordre s'effectue avec une energie d'activation de 24 kcal/mole.

Des cristaux de KZr2(AsO4)3 ont ete prepares par la methode de flux. L'etude cristallographique de ce compose a montre que celui-ci cristallise dans le systeme rhomboedrique avec les parametres a = 9,028(1) A; c = 24,399(3) A. d = 3,694 g/cm3; Z = 6 groupements formulaires par maille, le groupe d'espace est R3c. La structure a ete resolue par isotypie avec NaZr2(PO4)3 jusqu'a R = 0,041, pour 353 reflexions independantes, cette structure consiste en un enchainement tridimensionnel de tetraedres AsO4 et d'octaedres ZrO6; le potassium se trouve au centre d'un antiprisme avec des distances K—O egales a 2,813 A. Les distances Zr—O et As—O sont celles couramment observes As—O = 1,66 A et Zr—O = 2,05 A. Crystal Structure of KZr2(AsO4)3 KZr2(AsO4)3 crystals have been prepared by the flux method. The compound crystallizes in the rhombohedral system with the parameters: a = 9.028(1), c = 24.399(3) A. The density is d = 3.694 g/cm3 and Z = 6. The space group is R3c. The structure has been solved by isotypism with that of NaZr2(PO4)3 to R = 0.041, for 353 independent reflections. It consists in 3-dimensional chains of AsO4 tetrahedra and ZrO6 octahedra. K+ ions are the center of the antiprism with K—O distances equal to 2.813 A. The Zr–O and As–O distances have the same values of those usually observed (Zr–O = 2,05 A and As–O = 1.66 A).

Shen Yongfang, Durand Pierre-henri. Cinq ans d'acquisitions à la bibliothèque de l'Institut des Hautes Études Chinoises (1996-2000). In: Études chinoises, vol. 19, n°1-2, Printemps-Automne 2000. Mélanges de sinologie offerts à Jean-Pierre Diény ( II) pp. 311-323.

Pierre-Henri Durand : The Benevolent and the Mountain. A Three Steps Contribu¬ tion to the Study of Modern Prose. Dai Mingshi, famous for being the victim of the great literary case of Kangxi 's reign, has been, even before his success in imperial examinations, one of the most widely-spread author of modern prose of his time. This article takes the translation of the examination-paper which ranked Dai Mingshi first laureate at the 1709 metropolitan examination as the starting point to present an example of a genre most well-known as eight-legged essay (baguwen). Avoiding usual clichés and over-simplification this article puts emphasis on the variety, the complexity, and the richness of the exercise. Step by step, a close reading of the text attempts to define the style and concentrate on the substance to reconstruct the mosaic of inputs. The process helps to identify in modern prose characteristics close to our Western classical rhetoric., Dai Mingshi ne fut pas seulement la victime du grand procès littéraire du règne de Kangxi, il fut aussi, avant même de s'illustrer dans les concours impériaux, l'un des auteurs de prose moderne les plus répandus de son temps. La traduction de l'une des pièces qui lui valurent la première place à l'examen métropolitain de 1709 est le point de départ d'une présentation délaissant les simplifications des clichés habituels pour s'attacher à montrer la variété et la complexité, sinon la richesse, d'un genre plus connu sous le nom de prose à huit jambes (baguwen). Un retour au texte s'efforce d'en démêler la forme et d'en pénétrer le fond, pas à pas, point après point, reconstituant la mosaïque de ses emprunts, et reconnaissant dans la prose moderne des traits qui évoquent les sermons de notre rhétorique classique., Durand Pierre-henri. L'homme bon et la montagne : Petite contribution en trois temps à l'étude de la prose moderne. In: Études chinoises, vol. 18, n°1-2, Printemps-Automne 1999. Mélanges de sinologie offerts à Jean-Pierre Diény ( I) pp. 223-288.

Introduction: Lactobacillus plantarum 299v is a probiotic strain indicated to treat IBS patients. No other trial so far has tested the symptomatic effect of this strain in such a large population sample. Aims & methods: The aim of this multicenter double blind randomized placebocontrolled trial was to determine the efficacy of L.plantarum 299v in a powerful trial. Patients from 19 to 70 years old and suffering from chronic IBS according to Rome III criteria were recruited in 3 Indian centers. Treatment duration was 4 weeks long with per day one cap of placebo or one cap of 1010 cfu of L.plantarum 299v. Both frequency and intensity of abdominal pain, bloating, feeling of incomplete rectal evacuation and the number of stools were assessed by a VAS. The results are expressed as the percentage of improvement between baseline and the end of the treatment period. Overall patients' assessment was expressed on a 4-point rated scale. Significance analyses were made by using Chi-square or ANOVAR tests. Results: The ITT population comprised 200 patients and only 10 patients dropped out (PP population: 190). Neither the demographic characteristics nor the diet were different between the two treatment arms (see table 1). At the end of treatment, L.plantarum 299v improved significantly the symptoms both compared to baseline and to placebo (see table 2). Overall patients' assessment was significantly better in the L.plantarum 299v group compared to the placebo group (see table 2). Pulse and respiratory rates, blood pressure and internal temperature remained normal throughout the study without any significant changes in both arms. No side-effect was reported in both groups. Conclusion: A 4 weeks treatment with Lactobacillus plantarum 299v has shown to be very effective for the relief of symptoms, particularly abdominal pain and bloating in IBS patients fulfilling the Rome III criteria. Table 2: Clinical Evaluation and Patients' assessment

A transformation system for the thermophilic cellulolytic fungus Talaromyces sp. CL240 has been developed, using the phleomycin resistance gene from Streptoalloteichus hindustanus (Sh ble) as a dominant selectable marker. The plasmids (pAN8-1 and pUT720) carrying the Sh ble gene under the control of the Aspergillus nidulans glyceraldehyde-3-phosphate dehydrogenase (gpd) promoter, allowed selection of phleomycin-resistant transformants. A new promoter sequence cloned from chromosomal DNA of Trichoderma reesei (pUT737) was also able to drive efficient expression of the Sh ble gene in Talaromyces sp. CL240, resulting in the selection of transformants that were highly resistant to phleomycin.

Tolypocladium geodes strain NC50 was transformed by different integrating vectors bearing both a synthetic gene encoding human lysozyme (HLz) and the Sh ble phleomycin resistance marker, either in separate expression cassettes or in transcriptional or translational fusion configurations. Clones derived from all vectors were able to secrete HLz. The highest productivities in shake flasks (up to 150 mg l−1 in 5 days) were obtained when HLz was fused at the C-terminal end of the Sh ble protein. The fusion protein is efficiently secreted and release of active lysozyme occurs by extracellular proteolytic cleavage in the junction peptide.

International audience

Secreted heterologous proteins have been found to be produced much less efficiently by fungi than secreted homologous ones. This could be due, at least in part, to proteolytic cleavage by site-specific endoproteases of the secretory pathway, similar to the yeast KEX2 protease and the mammalian dibasic endoproteinases found in secretory pathways. Mature secreted fungal proteins may be protected from such cleavage due to the absence of cleavable sites in exposed regions. A comparison of the dipeptide distributions of 33 secreted and 34 cytoplasmic proteins from fungal producers of extracellular enzymes indicated a significant bias for some doublets, including the basic dipeptides Lys-Arg, Arg-Arg and Arg-Lys which have also been demonstrated to be KEX2 substrates. Other combinations were also found to be rare in secreted proteins, which could indicate either a broader specificity of the considered endopeptidase, or the presence either in the secretory organelles or among the secreted proteins of additional proteases with different specificities. Experimental evidence that the Lys-Arg site is processed in Tolypocladium geodes was provided by cloning a synthetic prosequence upstream of a phleomycin resistance (Sh ble) gene and analyzing the N-terminus of the corresponding protein purified from the culture supernatant. This system also provides a tool for further studies of specific proteases of fungi.

Penicillium occitanis strain Pol6, a mutant developed for hyperproduction of cellulase and pectinase enzymes was used for the study of extracellular pectinase production when pectins from different sources (apple and citrus) and with varying degree of esterification (DE) were used as inducers. Highly esterified citrus pectins were found to be suitable substrates for polygalacturonase, pectinase and pectin methyl esterase production, while low esterified citrus pectin favoured pectin lyase (PL) production. Apple pectins induced other hydrolytic enzymes (e.g., β-1,3-glucanase, β-glucosidase, β-galactosidase), in addition to pectolytic enzymes. Moreover, the combination of high and low esterified citrus pectins induced the production of a complete pectinase complex. The extent of degradation of the substrate and the affinity for PL decreased with decreasing DE irrespective of the source. There was no evidence of PL activity in this strain. No significant effect of cations (Ca++, Mn++, Na+) on PL activity was observed. However, EDTA (100 mm) inhibited 50% of the activity, when tested on highly esterified (rapid set citrus) pectin.

The enzymes produced by two thermophilic fungi claimed to produce heat-stable cellulases [see 1,4-(1,3;1,4)-β- d -glucan 4-glucanohydrolase, EC 3.2.1.4] have been compared to those of two mesophilic fungi on the basis of the following criteria: polysaccharolytic spectrum, heat and pH effects on stability and on activity of the different enzymes, and the ability to hydrolyse raw natural substrates. The cellulases produced by one of the thermophiles, Sporotrichum cellulophilum, appeared to be as heat-labile as those from the mesophile Trichoderma reesei ; moreover, the former enzyme preparation is the least efficient of the four tested. Thielavia terrestris enzymes are the most thermostable; on the basis of the other properties tested, T. terrestris enzymes are comparable to, or in some cases better than, those from mesophilic strains. However, the differences are not so great as to compensate for the much lower productivity of T. terrestris compared to the improved T. reesei and Penicillium sp. strains.

High-pressure liquid chromatography (HPLC) should be used more frequently in biomedical research because heavy molecules can be analysed at ambient temperature without derivatisation to volatile compounds. In this paper the separation of four free 17-ketosteroids, their ester sulfates and glucuronides by reversed-phase HPLC on a Micropak CH column is described. In order to improve the separation several parameters have been studied. Retention data allow comparison of the hydrophilic behaviour of free and conjugated steroids.

A strategy based on plate screening tests was designed for the selection of mutant strains of the fungus Trichoderma reesei suitable for cellulase (EC 3.2.1.4) production on an industrial scale. Six mutant generations were successively isolated, each of them fulfilling all of the three criteria: (1) improved productivity compared to the previous one, (2) high stability, (3) ability to be further improved. The mutant ultimately selected, CL 847, exhibited a four-fold increase in cellulase productivity in cellulose media as compared to the starting strain QM 9414, resistance to catabolite repression, increased β- d -glucosidase (EC 3.2.1.21) specific activity and it was partially constitutive. High cellulase concentrations were obtained in simple media with lactose or glucose as sole carbon source. The strain is currently used in an industrial fermentation plant.

The crystal structure of potassium difluorodiphosphate K2P2O5F2′ has been determined by direct methods and refined by a least-squares procedure using anisotropic thermal parameters. The R final index is 0.053 for 991 observed reflexions. The space group is C2 c : a = 12.614(8) A; b = 7.585(5) A; c = 7.195(5) A; β = 90.91(8)°, and Z = 4. The dimensions and configuration of the difluorodiphosphate ion have been determined.

L'effet de la nature du substituant R sur l'equilibre conformationnel s-cis-s-trans est examine dans la serie (CH3)2NCHCHCOR. L'utilisation d'un programme de calcul des courbes theoriques de l'echange permet la determination des parametres d'activation ΔH* et ΔS*. Restricted rotation around the CC bond in the OCCCN system is responsible of the s-cis and s-trans conformers as shown by the NMR spectra of vinylogous amides (CH3)2NCHCHCOR. Substituent effect (nature of R) is discussed in terms of conformational equilibrium. Theoretical line shapes have been computed and fit reasonably well with the experimental spectra; activation functions were also derived.

The protein coded by a bleomycin-resistance gene (ble) cloned from producing actinomycetes was purified from a culture of a recombinant E.coli strain and its action on bleomycin was determined by in vitro assays. The protein binds reversibly in a one to one ratio to bleomycin which can no longer cleave DNA. The bleomycin resistance of cells harboring a ble gene could be accounted for by a sequestering effect of the bleomycin-binding protein.

Après avoir caractérisé les zones arides par la faiblesse et l'imélgularité des précipitations et la forte évaporation, l'auteur décrit rapidement les pays du Sahel et leur milieu naturel. L'extrême rigueur des conditions de milieu n'autorise que l'agriculture pluviale ou l'agriculture irriguée. L'élevage exploite les graminées poussant sur les sables en saison des pluies ou les plantes des thalwegs en saison sèche. Cette agriculture précaire subvient tout juste aux besoins des populations et, soumise aux aléas du olimat, un déficit hydrique peut entraîner une famine. C'est ce qui s'est passé en 1972-1973. L'analyse de la pluviométrie de 1959 à 1974 montre que la sécheresse a commencé vers 1970 et a atteint son maximum en 1973, année où l'isohyète 300 mm (limite inférieure de la culture du imil) passait au sud du Niger. En conséquence, les pâturages se sont rétrécis, obligeant les troupeaux à des déplacements de plus en plus longs au cours desquels les bêtes sont mortes de faim et d'épuisement. Toutes les cultures, mêmes irriguées, ont été touchées provoquant la famine pour les hommes et le bétail, sans que l'eau de boisson ait manqué. Un puissant exode vers les vallées moins défavorisées s'est produit, créant des problèmes humains. L'auteur indique enfin quelques moyens de pallier les effets d'un retour de la sécheresse : dans l'immédiat, constituer des stocks de vivres, renouvelés chaque année permettant aux secours de s'organiser et créer de nouveaux points d'eau. A moyen terme, prendre les mesures nécessaires pour : a) augmenter l'eau disponible ; b ) économiser l'eau en réduisant les pertes par évaporation, drainage, ruissellement ou excès d'irrigation ; c) introduire la culture d'espè¬ ces précoces à court cycle végétatif consommant moins d'eau. A long terme, l'OJVLM. cherche à créer un réseau d'alerte à la sécheresse, qui fonctionnerait au début des années 1980., Durand Jacques-Henri. A propos de la sécheresse et ses conséquences au Sahel. In: Cahiers d'outre-mer. N° 120 - 30e année, Octobre-décembre 1977. pp. 383-403.

Durand Pierre-henri. Mandchous et Chinois : l'empereur Kangxi et le procès du Nanshan ji. In: Études chinoises, vol. 7, n°1, Printemps 1988. pp. 65-108.

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