Conjugated linolenic acid (CLnA), conjugated linoleic acid (CLA) and other biohydrogenation intermediates in plasma and milk fat of cows fed raw or extruded linseed

Thirty lactating dairy cows were used in a 333 Latin-square design to investigate the effects of a raw or extruded blend of linseed and wheat bran (70:30) on plasma and milk fatty-acids (FA). Linseed diets, containing 16.6% linseed blend on a dry-matter basis, decreased milk yield and protein percentage. They decreased the proportions of FA with less than 18 carbons in plasma and milk and resulted in cis-9, cis-12, cis-15 18:3 proportions that were more than three and four times higher in plasma and milk, respectively, whereas cis-9, cis-12 18:2 proportions were decreased by 10–15%. The cis-9, trans-11, cis-15 18:3 isomer of conjugated linolenic acid was not detected in the milk of control cows, but was over 0.15% of total FA in the milk fat of linseed-supplemented cows. Similarly, linseed increased plasma and milk proportions of all biohydrogenation (BH) intermediates in plasma and milk, including the main isomer of conjugated linoleic acid cis-9, trans-11 18:2, except trans-4 18:1 and cis-11, trans-15 18:2 in plasma lipids. In milk fat, compared with raw linseed, extruded linseed further reduced 6:0–16:0 even-chain FA, did not significantly affect the proportions of 18:0, cis-9 18:1 and cis-9, cis-12 18:2, tended to increase cis-9, cis-12, cis-15 18:3, and resulted in an additional increase in the proportions of most BH intermediates. It was concluded that linseed addition can improve the proportion of conjugated linoleic and linolenic acids, and that extrusion further increases the proportions of intermediates of ruminal BH in milk fat

Cis-15 intermediates of biohydrogenation in the duodenal flow of cows receiving linseed

Duodenal samples from a previous experiment (Akraim et al., 2006) were reanalysed for determination of cis-15 intermediates of ruminal biohydrogenation (BH). Three dry Holstein cows fitted with a duodenal cannula received a diet with 14% (dry matter basis) raw linseed. C18 fatty acids (C18FA) represented 6.3% of dietary dry matter, and contained 57.8% of C18:3. After 18 days adaptation, on each cow, twelve samples of duodenal flow were taken over 3 days, composited, and analysed for FA composition. Cis-9,trans-11,cis-15C18:3 and trans-11,cis-15C18:2 (CLnA) were identified by GC-MS. The proportion of CLnA was low, in spite of the high C18:3 intake. The first step of C18:3 BH, isomerisation to CLnA, was extensive because the C18:3 proportion was 13.5 times lower in the duodenum than in the diet (Akraim et al., 2006). The extensive C18:3 BH and the low proportion of CLnA suggest a rapid ruminal reduction of this FA. The proportion of the product of this first reduction, trans-11,cis-15C18:2, was much higher than that of CLnA, but quite variable among cows. This suggests that the reduction of this FA was slower than that of CLnA, although this reduction is supposed to be due to a broad range of bacteria. According to responsible bacteria, this reduction can lead to trans-11C18:1, or cis-15C18:1 and trans-15C18:1. Cis-15C18:1 represented only half the proportion of trans-15C18:1, suggesting either a direct BH of trans-11,cis-15C18:2 to trans-15C18:1, or a rapid isomerisation of cis-15C18:1 to trans-15C18:1

Déstabilisation gravitaire d'un matériau granulaire immergé

La stabilité gravitaire des ouvrages hydrauliques (digues fluviales, barrage en remblai, ...) est un problème complexifié par la présence d'infiltration d'eau au sein même de l'ouvrage et de ses fondations. Le territoire national compte près de 10000 km de digues, d'où un enjeu important en terme de risque d'inondation. Les déstabilisations gravitaires observées sur les ouvrages hydrauliques se distinguent par leur brièveté et par leur faible emprise spatiale. L'objectif de ce travail de thèse est d'étudier spécifiquement ces instabilités brèves et de faible déplacement cumulé le long de la pente dans le cas de matériaux granulaires saturés. Dans un premier temps, une campagne expérimentale a été effectuée sur la base d'un protocole de chargement progressif dans lequel un échantillon de sol est lentement incliné jusqu'à obtenir une déstabilisation massive. Il en ressort une phénoménologie complexe composée d'un nombre de précurseurs d'avalanche dépendant de la fraction volumique de l'échantillon. Une forte influence de la fraction volumique du sol sur le seuil de stabilité est également mise en avant. Une deuxième campagne expérimentale basée sur un protocole d'effondrement contrôlé, à inclinaison imposée, a été menée dans le but d'amplifier la cinématique de déstabilisation observée en chargement progressif. Mis à part les échantillons les plus denses qui produisent de simples avalanches de surface, on observe dans les autres cas un ou plusieurs événements successifs, d'allure circulaire, mobilisant des profondeurs très importantes contrairement aux avalanches de surface.. Outre l'inclinaison, on voit bien que, là encore, la fraction volumique est un paramètre clé dans le contrôle de la cinématique de déstabilisation. Des essais préliminaires laissent cependant entendre que, au-delà de la fraction volumique, c'est la microstructure qui semble être le vrai paramètre de contrôle à travers notamment la forme des grains et leur enchevêtrement.Gravitational stability of hydraulic structures (river dykes, embankment dam,...) is a problem complicated by the presence of water infiltration within the structure and its foundations. Our country has about 10,000 km of dikes and gravitational failure is a major challenge in terms of flood risk. In hydraulic structures, these destabilizations are characterized by their brevity and their small spatial extent. The objective of this thesis is to study specifically this type of brief instabilities in the case of a saturated granular material. A first series of experiments was carried out on the basis of a progressive loading protocol in which a soil sample is tilted slowly until a massive destabilization is triggered. It shows a phenomenology complex consisting of a number of precursory events prior final avalanching, dependent on the solid volume fraction of the sample. A strong impact of the solid volume fraction of the soil on the stability threshold is also highlighted. A second series of experiments based on a protocol of collapse controlled at imposed slope was conducted to amplify the kinematics of destabilization observed by progressive loading. Apart from the denser samples that produce simple surface avalanches, one or more successive events are observed in other cases. Unlike surface avalanches, these events mobilize grains much deeper, in an almost semi-circular area. In addition to slope angle, solid volume fraction is once again a key parameter in controlling the kinematics of destabilization. Preliminary tests suggest, however, that, beyond solid volume fraction, microstructure appears to be the true control parameter, particularly through grain shape and texture.SAVOIE-SCD - Bib.électronique (730659901) / SudocGRENOBLE1/INP-Bib.électronique (384210012) / SudocGRENOBLE2/3-Bib.électronique (384219901) / SudocSudocFranceF

Tassement d'un milieu granulaire : chargement surfacique et chargement volumique

Dans un sol granulaire immergé, un tassement du matériau peut être obtenu par différents types de chargement : un chargement en surface, par ajout d'un surpoids, ou un chargement en volume induit par un écoulement hydraulique interne. Cet article présente les prédictions théoriques dans chacune de ces deux situations en termes de tassement cumulé, de champ de déplacement et de profil vertical de compacité. La confrontation à des mesures expérimentales montre un accord satisfaisant avec la théorie

Detection of prions in the plasma of presymptomatic and symptomatic patients with variant Creutzfeldt-Jakob disease

Variant Creutzfeldt-Jakob disease (vCJD) is a human prion disease resulting from the consumption of meat products contaminated by the agent causing bovine spongiform encephalopathy. Evidence supporting the presence of a population of silent carriers that can potentially transmit the disease through blood transfusion is increasing. The development of a blood-screening assay for both symptomatic vCJD patients and asymptomatic carriers is urgently required. We show that a diagnostic assay combining plasminogen-bead capture and protein misfolding cyclic amplification (PMCA) technologies consistently detected minute amounts of abnormal prion protein from French and British vCJD cases in the required femtomolar range. This assay allowed the blinded identification of 18 patients with clinical vCJD among 256 plasma samples from the two most affected countries, with 100% sensitivity [95% confidence interval (CI), 81.5 to 100%], 99.2% analytical specificity (95% CI, 95.9 to 100%), and 100% diagnostic specificity (95% CI, 96.5 to 100%). This assay also allowed the detection of silent carriage of prions 1.3 and 2.6 years before the clinical onset in two blood donors who later developed vCJD. These data provide a key step toward the validation of this PMCA technology as a blood-based diagnostic test for vCJD and support its potential for detecting presymptomatic patients, a prerequisite for limiting the risk of vCJD transmission through blood transfusion

Using Pharmacokinetic and Viral Kinetic Modeling To Estimate the Antiviral Effectiveness of Telaprevir, Boceprevir, and Pegylated Interferon during Triple Therapy in Treatment-Experienced Hepatitis C Virus-Infected Cirrhotic Patients.: Effectiveness of triple therapy in cirrhotic patients

International audienceTriple therapy combining a protease inhibitor (PI) (telaprevir or boceprevir), pegylated interferon (PEG-IFN), and ribavirin (RBV) has dramatically increased the chance of eradicating hepatitis C virus (HCV). However, the efficacy of this treatment remains suboptimal in cirrhotic treatment-experienced patients. Here, we aimed to better understand the origin of this impaired response by estimating the antiviral effectiveness of each drug. Fifteen HCV genotype 1-infected patients with compensated cirrhosis, who were nonresponders to prior PEG-IFN/RBV therapy, were enrolled in a nonrandomized study. HCV RNA and concentrations of PIs, PEG-IFN, and RBV were frequently assessed in the first 12 weeks of treatment and were analyzed using a pharmacokinetic/viral kinetic model. The two PIs achieved similar levels of molar concentrations (P = 0.5), but there was a significant difference in the 50% effective concentrations (EC50) (P = 0.008), leading to greater effectiveness for telaprevir than for boceprevir in blocking viral production (99.8% versus 99.0%, respectively, P = 0.002). In all patients, the antiviral effectiveness of PEG-IFN was modest (43.4%), and there was no significant contribution of RBV exposure to the total antiviral effectiveness. The second phase of viral decline, which is attributed to the loss rate of infected cells, was slow (0.19 day(-1)) and was higher in patients who subsequently eradicated HCV (P = 0.03). The two PIs achieved high levels of antiviral effectiveness. However, the suboptimal antiviral effectiveness of PEG-IFN/RBV and the low loss of infected cells suggest that a longer treatment duration might be needed in cirrhotic treatment-experienced patients and that a future IFN-free regimen may be particularly beneficial in these patients

Assessment and validation of a suite of reverse transcription-quantitative PCR reference genes for analyses of density-dependent behavioural plasticity in the Australian plague locust

<p>Abstract</p> <p>Background</p> <p>The Australian plague locust, <it>Chortoicetes terminifera</it>, is among the most promising species to unravel the suites of genes underling the density-dependent shift from shy and cryptic solitarious behaviour to the highly active and aggregating gregarious behaviour that is characteristic of locusts. This is because it lacks many of the major phenotypic changes in colour and morphology that accompany phase change in other locust species. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) is the most sensitive method available for determining changes in gene expression. However, to accurately monitor the expression of target genes, it is essential to select an appropriate normalization strategy to control for non-specific variation between samples. Here we identify eight potential reference genes and examine their expression stability at different rearing density treatments in neural tissue of the Australian plague locust.</p> <p>Results</p> <p>Taking advantage of the new orthologous DNA sequences available in locusts, we developed primers for genes encoding 18SrRNA, ribosomal protein L32 (RpL32), armadillo (Arm), actin 5C (Actin), succinate dehydrogenase (SDHa), glyceraldehyde-3P-dehydrogenase (GAPDH), elongation factor 1 alpha (EF1a) and annexin IX (AnnIX). The relative transcription levels of these eight genes were then analyzed in three treatment groups differing in rearing density (isolated, short- and long-term crowded), each made up of five pools of four neural tissue samples from 5^thinstar nymphs. SDHa and GAPDH, which are both involved in metabolic pathways, were identified as the least stable in expression levels, challenging their usefulness in normalization. Based on calculations performed with the geNorm and NormFinder programs, the best combination of two genes for normalization of gene expression data following crowding in the Australian plague locust was EF1a and Arm. We applied their use to studying a target gene that encodes a Ca²⁺binding glycoprotein, <it>SPARC</it>, which was previously found to be up-regulated in brains of gregarious desert locusts, <it>Schistocerca gregaria</it>. Interestingly, expression of this gene did not vary with rearing density in the same way in brains of the two locust species. Unlike <it>S. gregaria</it>, there was no effect of any crowding treatment in the Australian plague locust.</p> <p>Conclusion</p> <p>Arm and EF1a is the most stably expressed combination of two reference genes of the eight examined for reliable normalization of RT-qPCR assays studying density-dependent behavioural change in the Australian plague locust. Such normalization allowed us to show that <it>C. terminifera </it>crowding did not change the neuronal expression of the <it>SPARC </it>gene, a gregarious phase-specific gene identified in brains of the desert locust, <it>S. gregaria</it>. Such comparative results on density-dependent gene regulation provide insights into the evolution of gregarious behaviour and mass migration of locusts. The eight identified genes we evaluated are also candidates as normalization genes for use in experiments involving other Oedipodinae species, but the rank order of gene stability must necessarily be determined on a case-by-case basis.</p

Neurodegeneration and neuroinflammation are linked, but independent of a-synuclein inclusions, in a seeding/spreading mouse model of Parkinson's disease

A key pathological process in Parkinson's disease (PD) is the transneuronal spreading of α-synuclein. Alpha-synuclein (α-syn) is a presynaptic protein that, in PD, forms pathological inclusions. Other hallmarks of PD include neurodegeneration and microgliosis in susceptible brain regions. Whether it is primarily transneuronal spreading of α-syn particles, inclusion formation, or other mechanisms, such as inflammation, that cause neurodegeneration in PD is unclear. We used a model of spreading of α-syn induced by striatal injection of α-syn preformed fibrils into the mouse striatum to address this question. We performed quantitative analysis for α-syn inclusions, neurodegeneration, and microgliosis in different brain regions, and generated gene expression profiles of the ventral midbrain, at two different timepoints after disease induction. We observed significant neurodegeneration and microgliosis in brain regions not only with, but also without α-syn inclusions. We also observed prominent microgliosis in injured brain regions that did not correlate with neurodegeneration nor with inclusion load. Using longitudinal gene expression profiling, we observed early gene expression changes, linked to neuroinflammation, that preceded neurodegeneration, indicating an active role of microglia in this process. Altered gene pathways overlapped with those typical of PD. Our observations indicate that α-syn inclusion formation is not the major driver in the early phases of PD-like neurodegeneration, but that microglia, activated by diffusible, oligomeric α-syn, may play a key role in this process. Our findings uncover new features of α-syn induced pathologies, in particular microgliosis, and point to the necessity for a broader view of the process of α-syn spreading