The application of NMR to study changes in polar metabolite concentrations in beef longissimus dorsi stored for different periods post mortem

In this study data generated by H-1 NMR spectroscopy were combined with chemometrics to analyse beef samples aged over a 21 day period. In particular, the amino acids, of which 12 were identified were found to increase over the ageing period with samples matured for 3 days having notably lower concentrations than carcasses aged for 21 days. This is believed to be a result of increased proteolysis within the muscle. This novel approach of using high resolution NMR spectrometry to analyse beef samples has not previously been reported and these findings demonstrate the potential of this technique linked with HPLC to be used as a suitable method for profiling meat samples.

Post-mortem examination of a wild muntjac from Northern Ireland.

1. Deer are of major concern with regards to impacts on biodiversity, forestry and agriculture as well as human health. The invasive Reeves’ muntjac deer Muntiacus reevesi, native to Asia but established in Great Britain, has recently appeared in the wild in Ireland.

2. The first verified record in the wild in Northern Ireland was confirmed during June 2009 as a result of a road traffic accident near Newtownards, Co. Down. The second record was a culled animal shot in the grounds of Mount Stewart, Co. Down during June 2011.

3. The current report aimed to perform a detailed investigation of the most recently obtained animal to establish a) its age, b) its genetic relationship with the first animal and c) the threat it might pose in terms of carrying endo- or ecto-parasites or other micro-pathogens, principally viruses and bacteria.

4. Analysis of its dentition suggested the culled animal was 56 weeks old (range 55-57 weeks).

5. Genetic analysis indicated that there was no possibility of a father-son relationship with the buck killed in 2009. There was a 6.25% probability of both individuals being full-sibs and a 25% probability of a half-sib relationship.

6. The deer was free from all the major pathogens. Most significantly, a novel species of Gammaherpesvirus was detected most closely matching type 2 ruminant rhadinovirus (Gammaherpesvirinae) from mule deer. Further sequencing is required to provide a definitive classification. There is no evidence suggesting this virus is pathogenic but its detection is nonetheless of concern.

7. During late 2009 (after the first animal was recovered) and early 2010, there were a number of sightings of muntjac within the vicinity of Mount Stewart. Our results indicate that the animal shot in June 2011 could not have been this same animal, as the shot animal must have been born in spring 2010. Moreover, genetic analysis indicates that the two recovered individuals were highly unlikely to share the same mother and father, suggesting they were the offspring of a minimum of three breeding adults (two fathers and one mother or two mothers and one father). This brings the total number of known individuals to 5 including the two offspring. The location of the breeding adults is unknown and may be either in captivity with subsequent escapees, or deliberate releases, or be free living in the wild. However, a further sighting during early 2012 may suggest a wild origin.

Metabolic signatures of human Alzheimer's disease (AD): H-1 NMR analysis of the polar metabolome of post-mortem brain tissue

Brain tissue from so-called Alzheimer's disease (AD) mouse models has previously been examined using H-1 NMR-metabolomics, but comparable information concerning human AD is negligible. Since no animal model recapitulates all the features of human AD we undertook the first H-1 NMR-metabolomics investigation of human AD brain tissue. Human post-mortem tissue from 15 AD subjects and 15 age-matched controls was prepared for analysis through a series of lyophilised, milling, extraction and randomisation steps and samples were analysed using H-1 NMR. Using partial least squares discriminant analysis, a model was built using data obtained from brain extracts. Analysis of brain extracts led to the elucidation of 24 metabolites. Significant elevations in brain alanine (15.4 %) and taurine (18.9 %) were observed in AD patients (p ≤ 0.05). Pathway topology analysis implicated either dysregulation of taurine and hypotaurine metabolism or alanine, aspartate and glutamate metabolism. Furthermore, screening of metabolites for AD biomarkers demonstrated that individual metabolites weakly discriminated cases of AD [receiver operating characteristic (ROC) AUC <0.67; p < 0.05]. However, paired metabolites ratios (e.g. alanine/carnitine) were more powerful discriminating tools (ROC AUC = 0.76; p < 0.01). This study further demonstrates the potential of metabolomics for elucidating the underlying biochemistry and to help identify AD in patients attending the memory clinic

mRNA Levels of BACE1 and its interacting proteins, RTN3 and PPIL2, correlate in human post mortem brain tissue

β-Site amyloid precursor protein cleaving enzyme (BACE1) is the rate-limiting enzyme for production of Aβ peptides, proposed to drive the pathological changes found in Alzheimer’s disease (AD). Reticulon 3 (RTN3) is a negative modulator of BACE1 (β-secretase) proteolytic activity, while peptidylprolyl isomerase (cyclophilin)-like 2 (PPIL2) positively regulated BACE1 gene expression in a cell-based assay. This study aimed to analyze RTN3 and PPIL2 mRNA levels in four brain regions from individuals with AD and controls. BACE1 mRNA had been previously quantified in the samples, as had glial fibrillary acidic protein (GFAP) and neuron-specific enolase (NSE), to track changing cell populations in the tissue. mRNA levels in the human post mortem brain tissue were assayed using quantitative real-time polymerase chain reaction (qPCR) and qbasePLUS, employing validated stably expressed reference genes. No differences in RTN3 or PPIL2 mRNA levels were found in individuals with AD, compared to controls. Both RTN3 and PPIL2 mRNA levels correlated significantly with BACE1 mRNA and all three showed similar disease stage-dependent changes with respect to NSE and GFAP. These findings indicated that the in vitro data demonstrating an effect of PPIL2 on BACE1 expression have functional relevance in vivo. Further research into BACE1-interacting proteins could provide a fruitful approach to the modulation of this protease and consequently Aβ production.

Multi-phase post-mortem CT angiography: development of a standardized protocol.

The objective of this work was to develop an easily applicable technique and a standardized protocol for high-quality post-mortem angiography. This protocol should (1) increase the radiological interpretation by decreasing artifacts due to the perfusion and by reaching a complete filling of the vascular system and (2) ease and standardize the execution of the examination. To this aim, 45 human corpses were investigated by post-mortem computed tomography (CT) angiography using different perfusion protocols, a modified heart-lung machine and a new contrast agent mixture, specifically developed for post-mortem investigations. The quality of the CT angiographies was evaluated radiologically by observing the filling of the vascular system and assessing the interpretability of the resulting images and by comparing radiological diagnoses to conventional autopsy conclusions. Post-mortem angiography yielded satisfactory results provided that the volumes of the injected contrast agent mixture were high enough to completely fill the vascular system. In order to avoid artifacts due to the post-mortem perfusion, a minimum of three angiographic phases and one native scan had to be performed. These findings were taken into account to develop a protocol for quality post-mortem CT angiography that minimizes the risk of radiological misinterpretation. The proposed protocol is easy applicable in a standardized way and yields high-quality radiologically interpretable visualization of the vascular system in post-mortem investigations.

Application of contrast media in post-mortem imaging (CT and MRI).

The application of contrast media in post-mortem radiology differs from clinical approaches in living patients. Post-mortem changes in the vascular system and the absence of blood flow lead to specific problems that have to be considered for the performance of post-mortem angiography. In addition, interpreting the images is challenging due to technique-related and post-mortem artefacts that have to be known and that are specific for each applied technique. Although the idea of injecting contrast media is old, classic methods are not simply transferable to modern radiological techniques in forensic medicine, as they are mostly dedicated to single-organ studies or applicable only shortly after death. With the introduction of modern imaging techniques, such as post-mortem computed tomography (PMCT) and post-mortem magnetic resonance (PMMR), to forensic death investigations, intensive research started to explore their advantages and limitations compared to conventional autopsy. PMCT has already become a routine investigation in several centres, and different techniques have been developed to better visualise the vascular system and organ parenchyma in PMCT. In contrast, the use of PMMR is still limited due to practical issues, and research is now starting in the field of PMMR angiography. This article gives an overview of the problems in post-mortem contrast media application, the various classic and modern techniques, and the issues to consider by using different media.

Les Signaux Post Mortem (SPM) de l’apoptose endothéliale : des acteurs du remodelage vasculaire

L’immunosuppression a permis d’améliorer l’incidence du rejet aigu sans toutefois améliorer significativement le rejet chronique. Celui-ci est caractérisé par une vasculopathie du greffon (VG) similaire à une forme accélérée d’athérosclérose native accompagnée de fibrose. La pathophysiologie de la VG découle de l’hypothèse de réponse à l’insulte proposée par Russell Ross en 1977. Selon son postulat, l’endothélium stressé par des facteurs immunologiques et non immunologiques initie l’apoptose endothéliale suivi d’une réponse de réparation vasculaire via un épaississement myo-intimal aux sites d’insultes. Toutefois, lorsque les stress endothéliaux initiaux demeurent soutenus, l’apoptose endothéliale et la réponse de réparation perpétuent. Compte tenu que l’inhibition de l’apoptose endothéliale bloque le développement de la VG in vivo, notre hypothèse de travail reposait sur les répercussions paracrines de l’apoptose endothéliale sur les types cellulaires participant au remodelage vasculaire. Nous avons généré un système expérimental in vitro afin d’induire l’apoptose endothéliale en absence significative de nécrose cellulaire. À l’aide d’une approche protéomique multidimensionnelle et comparative, nous avons démontré que les cellules endothéliales apoptotiques exportent spécifiquement 27 signaux post mortem (SPM). Nous avons démontré que certains de ces SPM ont des propriétés anti-apoptotiques (TCTP et EGF), d’autre fibrogénique (CTGF), récapitulant ainsi certains phénotypes cellulaires associés au développement de la VG. Parmi les médiateurs identifiés, 16 n’avaient pas de signal de sécrétion, incluant TCTP, suggérant que des mécanismes de sécrétion non conventionnels soient favorisés durant l’apoptose. Nous avons démontré que la caspase-3 effectrice régule la voie de sécrétion non classique exosomiale associée à l’export extracellulaire de nanovésicules TCTP+VE, anti-apoptotiques et biochimiquement distinctes des corps apoptotiques. Finalement, l’ensemble des données protéomiques ont permis d’émettre l’hypothèse qu’en réponse à un stress apoptotique, la cellule exporte différents médiateurs (solubles et vésiculaires) de manière non conventionnelle nécessitant la fusion d’organelles de la voie endocytaire et autophagique avec la membrane plasmique. Ce mécanisme serait régulé durant la phase effectrice de l’apoptose permettant ainsi d’initier une réponse de réparation extracellulaire seulement lorsque le destin cellulaire a atteint un point de non retour. Ainsi, le testament protéique et nanovésiculaire légué durant l’apoptose endothéliale pourrait servir simultanément de biomarqueur de la VG et de cible thérapeutique afin de diminuer le remodelage vasculaire pathologique.

Le portrait en post mortem immédiat des religieuses au Québec : influences, analyse stylistique et fortune graphique

Mémoire numérisé par la Division de la gestion de documents et des archives de l'Université de Montréal

Post-mortem des jeux de Vancouver de 2010 : retour sur la loi sur les marques olympiques et paralympiques

L’actualité entourant les Jeux olympiques de Vancouver de 2010 a fait couler beaucoup d’encre. Les retombées économiques enviables que procure l’exclusivité de la commandite aux Jeux olympiques suscitent la convoitise chez de nombreux concurrents commerciaux. Face aux phénomènes grandissant de la contrefaçon de la marque olympique et du marketing insidieux, le Comité international olympique et les pays hôtes des Jeux ont apporté une couche supplémentaire de protection pour la marque olympique. Au Canada, la Loi sur les marques olympiques et paralympiques (LMOP) fut sanctionnée le 22 juin 2007 par la Gouverneure Générale. Les sections-clés de cette loi sont arrivées à échéance le 31 décembre 2010 alors que d’autres de ses aspects sont permanents. La première partie de cet article a pour but de présenter les enjeux reliés à la protection de la propriété intellectuelle lors des Jeux de Vancouver, de définir et déterminer les limites de la propriété olympique, ainsi que le rôle de chaque acteur dans le domaine olympique. Nous étudierons les actions qui furent prises par le Comité d’organisation des Jeux olympiques et paralympiques d’hiver de 2010 à Vancouver (COVAN) et par les autres paliers de comités olympiques pour s’assurer la propriété de l’image olympique au Canada et l’importance de la LMOP dans leurs actions. Dans la deuxième partie de l’article, il sera question de la mise en œuvre de la protection de la marque olympique. Au cours des Jeux, le COVAN a dû faire face à l’adoption ou l’utilisation de la marque olympique par des tiers non autorisés et à des associations commerciales non autorisées avec l’image olympique. D’autres infractions ont également pris place sur Internet. Le présent article se veut un bilan de la situation canadienne suite aux Jeux olympiques de Vancouver de 2010. La LMOP a-t-elle été invoquée devant les tribunaux canadiens ? La loi a-t-elle effectivement été le seul rempart entre la protection olympique et les entreprises ou individus qui souhaitaient profiter indûment de l’événement ? Et finalement, cette initiative a-t-elle été déterminante dans le combat contre le marketing insidieux et autres attaques contre l’image olympique?

Tempus Post Mortem?* Adaptação portuguesa da transcendental-future time perspective scale (TFTPS)

Integrado no conceito mais geral da perspetiva temporal, o futuro transcendental tem sido conceptualizado como uma dimensão que abrange as crenças sobre o futuro a partir da morte imaginada do corpo físico até ao infinito. A transcendental-future time perspective scale (TFTPS) é uma escala unidimensional composta por 10 itens que avalia as cognições relacionadas com este espaço temporal. O objetivo deste estudo é apresentar a adaptação à língua e cultura portuguesa desta escala, assim como a sua estrutura fatorial e características psicométricas numa amostra de 346 participantes com idades compreendidas entre os 17 e os 54 anos (M = 19.87, DP = 4.27). Os resultados encontrados através da análise fatorial exploratória validaram a unidimensionalidade da escala (65.94 % de variância total explicada, α = 0.87).