Leishmaniose em Lobo Ibérico

Quando falamos de vida selvagem em Portugal, pensamos logo no Lobo Ibérico (Canis lupus signatus), que ao longo das últimas décadas tem vindo a sofrer um notório declínio populacional, apenas contrariado pelas medidas protecionistas entretanto implementadas. A diminuição do número de lobos em Portugal resulta principalmente da perseguição direta a que foram sujeitos e da destruição do seu habitat natural. Outra causa de redução/extinção de pequenas populações locais e fragmentadas de grandes carnívoros em outras partes no mundo tem sido as doenças infeciosas. Sendo monitorização da presença de patologias em animais silvestres fundamental no controle das zoonoses emergentes e na conservação das espécies. Neste contexto pretendeu-se elaborar um estudo de determinação da ocorrência de Leishmaniose, no Lobo Ibérico. Assim, recolheram-se aleatoriamente 42 amostras de sangue a lobos residentes no Parque Nacional Peneda-Gerês que, posteriormente foram testadas recorrendo-se ao método de ELISA. Através desta técnica, detetou-se que das 42 amostras testadas apenas uma amostra (2,4%) possuía anticorpos anti-leishmania. Um dos motivos para a obtenção de resultados pouco significativos poderá dever-se ao reduzido leque amostral. Concluímos, então, que são necessários estudos adicionais para avaliar a importância do Lobo Ibérico na transmissão e propagação da Leishmaniose.

The use of close-range photogrammetry in zooarchaeology : Creating accurate 3D models of wolf crania to study dog domestication

Acknowledgements We thank the Muséum National d'Histoire Naturelle, Paris, that provided access to the specimens, and access to the morphometric platform where the surface scans were performed. We also thank Raphael Cornette and Julien Claude for the fruitful discussions we had when writing the manuscript. This work was supported by NERC (grant number NE/K003259/1) and the European Research Council (ERC-2013-StG 337574-UNDEAD). This is publication ISEM 2016-127. We thank the two anonymous reviewers who greatly helped to improve the manuscript.

Les trois petits cochons et le grand méchant loup : stratégies visant la pérennité des populations de loups du Québec

L’objectif premier de cet essai est de prioriser des stratégies visant la pérennité des populations de loups au Québec. Ces stratégies visent à diminuer les pressions actuelles que subissent les loups. Les loups sont peu charismatiques. Il est donc difficile d’aller chercher l’appui du public. Si les loups du Québec parvenaient à gagner le cœur des Québécois, ils pourraient alors servir de levier afin que les dirigeants mettent en place un plan de conservation pour préserver ces espèces ou sous-espèces. C’est pourquoi d’ailleurs des stratégies sont proposées afin de rallier la population à cette cause. D’abord, un portrait des différentes espèces ou sous-espèces de loups et de coyotes est effectué pour bien comprendre dans quel contexte les loups évoluent actuellement. Ensuite, les menaces planant sur les populations sont identifiées, il s’agit de : l’hybridation avec le coyote ainsi que la compétition pour les ressources alimentaires, la fragmentation du territoire, les activités humaines, tels la chasse et le braconnage, les routes, ainsi que les activités touristiques. Les impacts économiques, sociaux et environnementaux reliés au déclin potentiel des loups sont ensuite énumérés. Cette partie démontre à quel point la situation est complexe et souligne toutes les répercussions potentielles que peut avoir la disparition du loup sur le Québec. Le rôle de l’appui du public dans le rétablissement d’espèces à statut précaire est ensuite discuté. Par la suite, une liste exhaustive des stratégies est mise en place. En tout, 43 stratégies ont été trouvées. Ces stratégies, visant l’atténuation des menaces sur le loup ou essayant de rallier l’appui du public, ont été classées dans 4 grandes orientations pour ensuite être priorisées selon leur performance. Par parcimonie, seules les 3 meilleures stratégies de chaque orientation seront présentées. Les 3 meilleures stratégies de l’orientation #1 sont de réduire le taux de mortalité des meutes de loups québécois, de ne pas clôturer les dépotoirs forestiers et de mettre au point un programme de suivi. Les 3 stratégies les plus performantes de l’orientation #2 sont de protéger les tanières, d’optimiser l’aménagement agricole et routier. Les 3 meilleures stratégies de l’orientation #3 sont d’utiliser la méthode de Fladry, de gérer les sorties guidées d’écoute nocturne et d’ajouter le loup de l’Est à la liste des espèces menacées du Québec. Les trois stratégies les plus performantes de l’orientation #4 sont d’informer la population de sa capacité à participer et à aider à conserver les loups, d’encourager les parcs et les réserves à inclure le loup dans leurs indicateurs de l’intégrité écologique des écosystèmes et d’informer la population sur les avantages découlant de la conservation des loups. Dans le cadre des recommandations, il faut aussi combiner diverses stratégies de différentes orientations, utiliser le marketing social sur la majorité des stratégies prioritaires, mener une campagne d’éducation populaire à propos du loup et inclure l’ensemble des parties prenantes pouvant avoir un impact sur les différentes stratégies. Pour terminer, l’intérêt d’introduire le loup sur l’île d’Anticosti est évoqué, car cette avenue pourrait être une solution intéressante afin de remédier aux problématiques qu’entraînent les fortes densités de cerfs de Virginie sur cette île.

Viabilidade e segurança do transplante intratecal de células-tronco mesenquimais autólogas e alogênicas provenientes da medula óssea de cães (lupus canis familiaris)

Pós-graduação em Medicina Veterinária - FMVZ

Targeting cancer with a lupus autoantibody

Systemic lupus erythematosus (SLE) is distinct among autoimmune diseases because of its association with circulating autoantibodies reactive against host DNA. The precise role that anti-DNA antibodies play in SLE pathophysiology remains to be elucidated, and potential applications of lupus autoantibodies in cancer therapy have not previously been explored. We report the unexpected finding that a cell-penetrating lupus autoantibody, 3E10, has potential as a targeted therapy for DNA repair–deficient malignancies. We find that 3E10 preferentially binds DNA single-strand tails, inhibits key steps in DNA single-strand and double-strand break repair, and sensitizes cultured tumor cells and human tumor xenografts to DNA-damaging therapy, including doxorubicin and radiation. Moreover, we demonstrate that 3E10 alone is synthetically lethal to BRCA2-deficient human cancer cells and selectively sensitizes such cells to low-dose doxorubicin. Our results establish an approach to cancer therapy that we expect will be particularly applicable to BRCA2-related malignancies such as breast, ovarian, and prostate cancers. In addition, our findings raise the possibility that lupus autoantibodies may be partly responsible for the intrinsic deficiencies in DNA repair and the unexpectedly low rates of breast, ovarian, and prostate cancers observed in SLE patients. In summary, this study provides the basis for the potential use of a lupus anti-DNA antibody in cancer therapy and identifies lupus autoantibodies as a potentially rich source of therapeutic agents.

Assessing the taxonomic status of dingoes Canis familiaris for conservation

1. The conservation status of the dingo Canis familiaris dingo is threatened by hybridization with the domestic dog C. familiaris familiaris. A practical method that can estimate the different levels of hybridization in the field is urgently required so that animals below a specific threshold of dingo ancestry (e.g. 1/4 or 1/2 dingoes) can reliably be identified and removed from dingo populations. 2. Skull morphology has been traditionally used to assess dingo purity, but this method does not discriminate between the different levels of dingo ancestry in hybrids. Furthermore, measurements can only be reliably taken from the skulls of dead animals. 3. Methods based on the analysis of variation in DNA are able to discriminate between the different levels of hybridization, but the validity of this method has been questioned because the materials currently used as a reference for dingoes are from captive animals of unproven genetic purity. The use of pre-European materials would improve the accuracy of this method, but suitable material has not been found in sufficient quantity to develop a reliable reference population. Furthermore, current methods based on DNA are impractical for the field-based discrimination of hybrids because samples require laboratory analysis. 4. Coat colour has also been used to estimate the extent of hybridization and is possibly the most practical method to apply in the field. However, this method may not be as powerful as genetic or morphological analyses because some hybrids (e.g. Australian cattle dog × dingo) are similar to dingoes in coat colour and body form. This problem may be alleviated by using additional visual characteristics such as the presence/absence of ticking and white markings.

Genetic studies on Finnish lupus erythematosus patients with cutaneous manifestations

Lupus erythematosus (LE) is a chronic, heterogeneous autoimmune disorder with abnormal immune responses, including production of autoantibodies and immune complexes. Clinical presentations of the disease range from mild cutaneous manifestations to a more generalised systemic involvement of internal organs. Cutaneous (CLE) forms are further subclassified into discoid LE (DLE), subacute cutaneous LE (SCLE) and acute cutaneous lupus erythematosus (ACLE), and may later progress to systemic disease (SLE). Both genetic and environmental factors contribute to the disease, although the precise aetiology is still elusive. Furthermore, complex gene-gene or gene-environment interactions may result in different subphenotypes of lupus. The genetic background of CLE is poorly known and only a few genes are confirmed, while the number of robust genetic associations in SLE exceeds 30. The aim of this thesis was to characterise the recruited patients clinically, and identify genetic variants conferring susceptibility to cutaneous variants of LE. Given that cutaneous and systemic disease may share underlying genetic factors, putative CLE candidate genes for genotyping were selected among those showing strong evidence of association in SLE. The correlation between relevant clinical manifestations and risk genotypes was investigated in order to find specific subphenotype associations. In addition, epistatic interactions in SLE were studied. Finally, the role of tissue degrading matrix metalloproteinases (MMP) in LE tissue injury was explored. These studies were conducted in Finnish case-control and family cohort, and Swedish case-control cohort. The clinical picture of the patients in terms of cutaneous, haematological and immunological findings resembled that described in the contemporary literature. However, the proportion of daily smokers was very high supporting the role of smoking in disease aetiology. The results confirmed that, even though clinically distinct entities, CLE and SLE share predisposing genetic factors. For the first time it was shown that known SLE susceptibility genes IRF5 and TYK2 also increase the risk of CLE. A tendency toward gene-gene interaction between these genes was found in SLE. As a remarkable novel finding, it was observed that ITGAM polymorphisms associated even more strongly to DLE than SLE, and the risk estimates were substantially higher than those reported for SLE. Several other recently identified SLE susceptibility genes showed signs of good or modest association especially in DLE. Subphenotype analyses indicated possible associations to clinical features, but marginally significant results reflected lack of sufficient power for these studies. Thorough immunohistochemical analyses of several MMPs demonstrated a role in epidermal changes and dermal tissue remodelling in diseased skin, and suggested that targeted action using selective MMP inhibitors may reduce lupus-induced damage in inflamed tissues. In conclusion, the results provide an insight into the genetics of CLE and demonstrate that genetic predisposition is at least in part shared between cutaneous and systemic variants of LE. This doctoral study has contributed IRF5, TYK2, ITGAM and several other novel genes to the so far short list of genes implicated in CLE susceptibility. Detailed examination of the function of these genes in CLE pathogenesis warrants further studies. Furthermore, the results support the need of subphenotype analysis with sample sizes large enough to reveal possible specific disease associations in order to better understand the heterogeneous nature and clinical specificities of the disease. Comprehensive analysis of clinical data suggests that smoking is an environmental triggering factor.

Ribosomal phosphoproteins in Microsporum canis

Ribosomal phosphoproteins of Microsporum canis labelled in vivo were characterised by two-dimensional and SDS polyacrylamide gel electrophoresis. A small subunit protein, S6, was the only phosphoprotein identified in 40S and 80S in basic-acidic two-dimensional gels. Three different forms of phosphorylated S6 were also observed in 40S subunit. On SDS gels five phosphoproteins were identified in 80S; of these three were present in 40S and two in 60S. S6 was the only basic phosphoprotein, while the other four were acidic.

IgG4 Autoantibodies to DNA in Systemic Lupus erythematosus Patients

The presence of DNA-specific IgG4 antibodies was demonstrated in the sera of patients with systemic lupus erythematosus (SLE) by a microtiter solid-phase radioimmunoassay. A patient with distal inter-phalangeal swelling and extensive ulcers in the oral cavity, seronegative for anti-DNA antibodies of the IgG isotype, was found to have anti-DNA autoantibodies exclusively of the IgG4 subclass. These autoantibodies directed against the dsDNA conformation cross-reacted with chondroitin sulfate, dermatan sulfate and heparin.