Dying neurons in thalamus of asphyxiated term newborns and rats are autophagic

Enjeu: Déterminer si la macroautophagie est activée de façon excessive dans les neurones en souffrance dans l'encéphalopathie anoxique-ischémique du nouveau-né à terme. Contexte de la recherche: L'encéphalopathie anoxique-ischémique suite à une asphyxie néonatale est associée à une morbidité neurologique à long terme. Une diminution de son incidence reste difficile, son primum movens étant soudain, imprévisible voire non identifiable. Le développement d'un traitement pharmacologique neuroprotecteur post-anoxie reste un défi car les mécanismes impliqués dans la dégénérescence neuronale sont multiples, interconnectés et encore insuffisamment compris. En effet, il ressort des études animales que la notion dichotomique de mort cellulaire apoptotique (type 1)/nécrotique (type 3) est insuffisante. Une même cellule peut présenter des caractéristiques morphologiques mixtes non seulement d'apoptose et de nécrose mais aussi parfois de mort autophagique (type 2) plus récemment décrite. L'autophagie est un processus physiologique normal et essentiel de dégradation de matériel intracellulaire par les enzymes lysosomales. La macroautophagie, nommée simplement autophagie par la suite, consiste en la séquestration de parties de cytosol à éliminer (protéines et organelles) dans des compartiments intermédiaires, les autophagosomes, puis en leur fusion avec des lysosomes pour former des autolysosomes. Dans certaines conditions de stress telles que l'hypoxie et l'excitoxicité, une activité autophagique anormalement élevée peut être impliquée dans la mort cellulaire soit comme un mécanisme de mort indépendant (autodigestion excessive correspondante à la mort cellulaire de type 2) soit en activant d'autres voies de mort comme celles de l'apoptose. Description de l'article: Ce travail examine la présence de l'autophagie et son lien avec la mort cellulaire dans les neurones d'une région cérébrale fréquemment atteinte chez le nouveau- né humain décédé après une asphyxie néonatale sévère, le thalamus ventro-latéral. Ces résultats ont été comparés à ceux obtenus dans un modèle d'hypoxie-ischémie cérébrale chez le raton de 7 jours (dont le cerveau serait comparable à celui d'un nouveau-né humain de 34-37 semaines de gestation). Au total 11 nouveau-nés à terme décédés peu après la naissance ont été rétrospectivement sélectionnés, dont 5 présentant une encéphalopathie hypoxique- ischémique sévère et 6 décédés d'une cause autre que l'asphyxie choisis comme cas contrôle. L'autophagie et l'apoptose neuronale ont été évaluées sur la base d'une étude immunohistochimique et d'imagerie confocale de coupes histologiques en utilisant des marqueurs tels que LC3 (protéine dont la forme LC3-II est liée à la membrane des autophagosomes), p62/SQSTM1 (protéine spécifiquement dégradée par autophagie), LAMP1 (protéine membranaire des lysosomes et des autolysosomes), Cathepsin D ou B (enzymes lysosomales), TUNEL (détection de la fragmentation de l'ADN se produisant lors de l'apoptose), CASPASE-3 activée (protéase effectrice de l'apoptose) et PGP9.5 (protéine spécifique aux neurones). Chez le raton l'étude a pu être étendue en utilisant d'autres méthodes complémentaires telles que la microscopie électronique et le Western-blot. Une quantification des différents marqueurs montre une augmentation statistiquement significative de l'autophagie neuronale dans les cas d'asphyxie par rapport aux cas contrôles chez l'humain comme chez le raton. En cas d'asphyxie, les mêmes neurones expriment une densité accrue d'autophagosomes et d'autolysosomes par rapport aux cas contrôles. De plus, les neurones hautement autophagiques présentent des caractéristiques de l'apoptose. Conclusion: Cette étude montre, pour la première fois, que les neurones thalamiques lésés en cas d'encéphalopathie hypoxique-ischémique sévère présentent un niveau anormalement élevé d'activité autophagique comme démontré chez le raton hypoxique-ischémique. Ce travail permet ainsi de mettre en avant l'importance de considérer l'autophagie comme acteur dans la mort neuronale survenant après asphyxie néonatale. Perspectives: Récemment un certain nombre d'études in vitro ou sur des modèles d'ischémie cérébrale chez les rongeurs suggèrent un rôle important de la macroautophagie dans la mort neuronale. Ainsi, l'inhibition spécifique de la macroautophagie devrait donc être envisagée dans le futur développement des stratégies neuroprotectrices visant à protéger le cerveau des nouveau-nés à terme suite à une asphyxie

Field evaluation of the CATT/Trypanosoma brucei gambiense on blood-impregnated filter papers for diagnosis of human African trypanosomiasis in southern Sudan.

Most Human African Trypanosomiasis (HAT) control programmes in areas endemic for Trypanosoma brucei gambiense rely on a strategy of active mass screening with the Card Agglutination Test for Trypanosomiasis (CATT)/T. b. gambiense. We evaluated the performance, stability and reproducibility of the CATT/T. b. gambiense on blood-impregnated filter papers (CATT-FP) in Kajo-Keji County, South-Sudan, where some areas are inaccessible to mobile teams. The CATT-FP was performed with a group of 100 people with a positive CATT on whole blood including 17 confirmed HAT patients and the results were compared with the CATT on plasma (CATT-P). The CATT-FP was repeated on impregnated filter papers stored at ambient and refrigerated temperature for 1, 3, 7 and 14 days. Another 82 patients with HAT, including 78 with a positive parasitology, were tested with the CATT-FP and duplicate filter paper samples were sent to a reference laboratory to assess reproducibility. The CATT-FP was positive in 90 of 99 patients with HAT (sensitivity: 91%). It was less sensitive than the CATT-P (mean dilution difference: -2.5). There was no significant loss of sensitivity after storage for up to 14 days both at ambient and cool temperature. Reproducibility of the CATT-FP was found to be excellent (kappa: 0.84). The CATT-FP can therefore be recommended as a screening test for HAT in areas where the use of CATT-P is not possible. Further studies on larger population samples in different endemic foci are still needed before the CATT-FP can be recommended for universal use

Thermic effect of glucose and amino acids in man studied by direct and indirect calorimetry

1. In order to reinvestigate the classical concept of specific dynamic action of food, the thermic effect of ingested glucose (50 g) or essential amino acids (50 g) or both was measured in seven healthy male subjects dressed in shorts, by using both direct and indirect calorimetry simultaneously. Experiments were performed under conditions of thermal comfort at 28°. 2. Energy ‘balance' (heat production minus heat losses) was negative during the control period (mean heat deficit: −16.0 ± 0.8 kJ/m2 per h. 3. Metabolic rate increased 13.6 ± 1.8% after the glucose load, 17.2 ± 1.4% after amino acids, and 17.3 ± 2.9% after both glucose and amino acids: thus there was no additive thermic effect when both nutrients were given together. 4. In contrast to the metabolic rate, heat losses were not significantly altered after nutrient ingestion; consequently, the energy ‘balance' became rapidly positive. 5. These results show that: (a) the food-induced thermogenesis, for a moderate energy intake, is less dependent on the nature of the nutrients than was classically admitted; (b) this increased heat production mainly induces changes in heat storage rather than in heat losses during the first hours following ingestion of a mea

P298 Comparison between clinical and patient-reported symptoms among Crohn's disease and ulcerative colitis patients

Background: There is no symptom-based patient-reported outcomes (PRO) measurement available in IBD. Disease scores contain a mixture of PRO and physician's observations and have shown serious limitations in clinical trials. Comparison between healthcare professionals (HCP) and patient (P) reports on scores' items is a first step toward disease scores refinement. In our IBD cohort study, we were able to collect P and HCP-reported symptoms independently. We assessed the agreement between both measures, and tested the correlation between the general well-being item (GWB) and two health-related quality of life (HRQoL) measures. Methods: Between 2012 and 2015, we collected CDAI and MTWAI items 1) during follow-up medical visits, 2) through P self-reported follow-up questionnaire, except lab values. We compared items independently reported by HCP and P, stratified by diagnostic and Δt HCP-P reports. We calculated the Cohen's kappa (κ) statistic for agreement. A quadratic weight was applied for more severely serious disagreements. For EIM & complications, we computed a pooled κ based on the average between observed and expected probability of agreement over sub-items. A pooled κ was computed to summarize agreement over all examined variables. We also collected SF-36 and IBDQ scores. Pearson correlation coefficients r were calculated between both scores and GWB reports of HCP and P. Results: 2427 reports could be evaluated (Δt: 537<1 month, 390 1–2, 1500 2–6), referring to 1385 patients (52% females, 58% CD)

Earth current monitoring circuit for inductive loads

The search for higher magnetic fields in particle accelerators increasingly demands the use of superconducting magnets. This magnet technology has a large amount of magnetic energy storage during operation at relatively high currents. As such, many monitoring and protection systems are required to safely operate the magnet, including the monitoring of any leakage of current to earth in the superconducting magnet that indicates a failure of the insulation to earth. At low amplitude, the earth leakage current affects the magnetic field precision. At a higher level, the earth leakage current can additionally generate local losses which may definitively damage the magnet or its instrumentation. This paper presents an active earth fault current monitoring circuit, widely deployed in the converters for the CERN Large Hadron Collider (LHC) superconducting magnets. The circuit allows the detection of earth faults before energising the circuit as well as limiting any eventual earth fault current. The electrical stress on each circuit component is analyzed and advice is given for a totally safe component selection in relation to a given load

Perioperative Antibiotic Prophylaxis of Wound and Foreign Body Infections: Microbial Factors Affecting Efficacy

Numerous microbial factors are responsible for perioperative infections and influence the efficacy of antibiotic prophylaxis. These factors include the staphylococcal carrier state, bacterial adherence to a number of host proteins, the production of glycocalyx by sessile bacteria, and shifts in antibiotic resistance. A full understanding of the mechanisms involved will lead to further reductions in the number of postoperative infections. Unfortunately, the microbial factors affecting prophylaxis cannot be evaluated separately under clinical conditions; they are easier to study under circumstances whose bacteriologic features are well defined and in which the presence of foreign materials (e.g., sutures) greatly potentiates pathogenic mechanisms. Such circumstances exist, for example, in infections developing after "clean” surgery and in experimental models. Since even clean wounds are found to be contaminated when sampled carefully, the control of infection is more a quantitative than a qualitative problem. The critical period for the development of infection is short: an antibiotic course not exceeding 24 hours seems effective in preventing infectio

Microbiological Factors Influencing the Outcome of Nosocomial Bloodstream Infections: A 6-Year Validated, Population-Based Model

All patients (n = 1,745) with nosocomial bloodstream infection identified between 1986 and 1991 at a single 900-bed tertiary care hospital were studied to identify microbiological factors independently associated with mortality due to the infection. Patients were identified by prospective, case-based surveillance and positive blood cultures. Mortality rates were examined for secular trends. Prognostic factors were determined with use of univariate and multivariate analyses, and both derivation and validation sets were used. A total of 1,745 patients developed nosocomial bloodstream infection. The 28-day crude mortality was 22%, and crude in-hospital mortality was 35%. Factors independently (all P < .05) associated with increased 28-day mortality rates were older age, longer length of hospital stay before bloodstream infection, and a diagnosis of cancer or disease of the digestive system. After adjustment for major confounders, Candida species were the only organisms independently influencing the outcome of nosocomial bloodstream infection (odds ratio [OR] for mortality = 1.84; 95% confidence interval [CI], 1.22-2.76; P = .0035). The two additional microbiological factors independently associated with increased mortality were pneumonia as a source of secondary infection (OR = 2.74; 95% CI, 1.87-4.00; P < .0001) and polymicrobial infection (OR = 1.68; 95% CI, 1.22-2.32; P = .0014). Our data suggest that microbiological factors independently affect the outcome of nosocomial bloodstream infectio