Analysis of translation of 5’ untranslated regions in cancer

Short upstream open reading frames (uORFs) are cis-acting elements located within the 5'-leader sequence of transcripts. Recent genome-wide ribosome profiling (RiboSeq) studies have demonstrated the widespread presence of uORFs in the transcriptome and have shown that many uORFs can initiate with non-AUG codons. uORFs can impact gene expression of the downstream main open reading frame (mORF) by triggering messenger RNA (mRNA) decay or by regulating translation. Thus, disruption, elimination or creation of uORFs can elicit the development of several genetic diseases, such as cancer. The ATP-binding cassette subfamily E member 1 (ABCE1) gene belongs to the ABC gene transporter superfamily. However, it does not behave as a drug transporter like the other members of this family. ABCE1 actively participates in the different stages of the translation process and is involved in cell proliferation and anti-apoptotic signaling processes, associating ABCE1 to a potential oncogenic function. RiboSeq occupancy profiles of the ABCE1 mRNA 5’-leader sequence indicate an active translation associated with the presence of uORFs, which is suggestive of a high translational regulation. Our aim was to study the translational regulation mediated by the five AUG and five non-AUG uORFs present in the human ABCE1 5’-leader sequence in colorectal cancer. With this purpose, we constructed a set of Firefly luciferase (FLuc) reporter vectors derived from the wild-type one containing the native configuration of the human ABCE1 5’-leader sequence upstream of the FLuc ORF, and transiently transfected colorectal cancer HCT116 cells. Here we show that ABCE1 mORF expression is regulated by its uORFs. Our results are consistent with a model wherein uORF1 recruits ribosomes onto the mRNA, behaving like a ribosomal barrier. The ribosomes that efficiently bypass uORF1 and/or uORF2, must probably reinitiate at uORF3 and/or uORF5, while uORF4 is greatly bypassed. uORF3 and uORF5 function as repressive uORFs that may cooperate to reach a maximum repression of the mORF. Thus, both bypass and reinitiation events of the AUG uORFs within ABCE1 5’-leader sequence contribute for the translational control of the mORF. In constrast, the non-AUG uORFs seem to be devoid of a significant inhibitory activity. The AUG uORF-mediated translational control is maintained in normal and in endoplasmic reticulum (ER) stress conditions, which keeps the expression level of ABCE1 at a minimum, showing that ABCE1 is a stress-resistant transcript whose functions are equally essential in normal and in coditions of global translation impairment. In addition, we show that ABCE1 uORF-mediated translational regulation is preserved in non-tumorigenic and cancerous cells, which is consistent with a lack of an oncogenic function by the uORFs, as well as ABCE1 itself, in the colorectal cancer cell line tested. This study contributes with an additional example of how uORF-mediated translational regulation can occur. In addition, it reveals how important is to screen the 5’-leader sequence of the transcripts in search for potential disease-related variants. This information might be relevant for the implementation of new diagnostic and/or therapeutic tools for diseases associated with the deregulation of uORF-mediated translational control.BioISI – Biosystems & Integrative Sciences Institute da Faculdade de Ciências da Universidade de Lisboa (UID/MULTI/04046/2013

Caracterização do potencial citotóxico e mecanismos de acção de um complexo de platina de configuração trans em células animais

Dissertação apresentada para obtenção do Grau de Mestre em Genética Molecular e Biomedicina, pela Universidade Nova de Lisboa, Faculdade de Ciências e Tecnologi

Preclinical assessment of mesenchymal-stem-cell-based therapies in spinocerebellar ataxia type 3

The low regeneration potential of the central nervous system (CNS) represents a challenge for the development of new therapeutic strategies for neurodegenerative diseases, including spinocerebellar ataxias. Spinocerebellar ataxia type 3 (SCA3)—or Machado–Joseph disease (MJD)—is the most common dominant ataxia, being mainly characterized by motor deficits; however, SCA3/MJD has a complex and heterogeneous pathophysiology, involving many CNS brain regions, contributing to the lack of effective therapies. Mesenchymal stem cells (MSCs) have been proposed as a potential therapeutic tool for CNS disorders. Beyond their differentiation potential, MSCs secrete a broad range of neuroregulatory factors that can promote relevant neuroprotective and immunomodulatory actions in different pathophysiological contexts. The objective of this work was to study the effects of (1) human MSC transplantation and (2) human MSC secretome (CM) administration on disease progression in vivo, using the CMVMJD135 mouse model of SCA3/MJD. Our results showed that a single CM administration was more beneficial than MSC transplantation—particularly in the cerebellum and basal ganglia—while no motor improvement was observed when these cell-based therapeutic approaches were applied in the spinal cord. However, the effects observed were mild and transient, suggesting that continuous or repeated administration would be needed, which should be further tested.This research was funded by the National Ataxia Foundation (NAF) and by Portuguese national funds, through the Foundation for Science and Technology (FCT)—projects UIDB/50026/2020, UIDP/50026/2020, POCI-01-0145-FEDER-029206, and through the Santa Casa Neuroscience Awards (Santa Casa da Misericórdia Lisboa)—project MC-04/17. Additionally, this project was funded by the ICVS Scientific Microscopy Platform, a member of the national infrastructure PPBI—Portuguese Platform of Bioimaging (PPBI-POCI-01-0145-FEDER-022122). S.C.S. received an individual fellowship within the project TUBITAK/0007/2014. The FCT funded individual fellowships to J.S C., A.N.-C., B.M.- P., F.G.T., R.L., S.M., N.A.S., C.S.-C., and S.D.-S. (SFRH/BD/140624/2018, SFRH/BPD/118779/2016, SFRH/BD/120124/2016, SFRH/BPD/118408/2016, PD/BDE/127836/2016, CEECIND/01902/2017, CEECIND/04794/2017, CEECIND/03887/2017, and CEECIND/00685/2020)

MAMMALS IN PORTUGAL : A data set of terrestrial, volant, and marine mammal occurrences in P ortugal

Mammals are threatened worldwide, with 26% of all species being includedin the IUCN threatened categories. This overall pattern is primarily associatedwith habitat loss or degradation, and human persecution for terrestrial mam-mals, and pollution, open net fishing, climate change, and prey depletion formarine mammals. Mammals play a key role in maintaining ecosystems func-tionality and resilience, and therefore information on their distribution is cru-cial to delineate and support conservation actions. MAMMALS INPORTUGAL is a publicly available data set compiling unpublishedgeoreferenced occurrence records of 92 terrestrial, volant, and marine mam-mals in mainland Portugal and archipelagos of the Azores and Madeira thatincludes 105,026 data entries between 1873 and 2021 (72% of the data occur-ring in 2000 and 2021). The methods used to collect the data were: live obser-vations/captures (43%), sign surveys (35%), camera trapping (16%),bioacoustics surveys (4%) and radiotracking, and inquiries that represent lessthan 1% of the records. The data set includes 13 types of records: (1) burrowsjsoil moundsjtunnel, (2) capture, (3) colony, (4) dead animaljhairjskullsjjaws, (5) genetic confirmation, (6) inquiries, (7) observation of live animal (8),observation in shelters, (9) photo trappingjvideo, (10) predators dietjpelletsjpine cones/nuts, (11) scatjtrackjditch, (12) telemetry and (13) vocalizationjecholocation. The spatial uncertainty of most records ranges between 0 and100 m (76%). Rodentia (n=31,573) has the highest number of records followedby Chiroptera (n=18,857), Carnivora (n=18,594), Lagomorpha (n=17,496),Cetartiodactyla (n=11,568) and Eulipotyphla (n=7008). The data setincludes records of species classified by the IUCN as threatened(e.g.,Oryctolagus cuniculus[n=12,159],Monachus monachus[n=1,512],andLynx pardinus[n=197]). We believe that this data set may stimulate thepublication of other European countries data sets that would certainly contrib-ute to ecology and conservation-related research, and therefore assisting onthe development of more accurate and tailored conservation managementstrategies for each species. There are no copyright restrictions; please cite thisdata paper when the data are used in publications.info:eu-repo/semantics/publishedVersio

Mammals in Portugal: a data set of terrestrial, volant, and marine mammal occurrences in Portugal

Mammals are threatened worldwide, with ~26% of all species being included in the IUCN threatened categories. This overall pattern is primarily associated with habitat loss or degradation, and human persecution for terrestrial mammals, and pollution, open net fishing, climate change, and prey depletion for marine mammals. Mammals play a key role in maintaining ecosystems functionality and resilience, and therefore information on their distribution is crucial to delineate and support conservation actions. MAMMALS IN PORTUGAL is a publicly available data set compiling unpublished georeferenced occurrence records of 92 terrestrial, volant, and marine mammals in mainland Portugal and archipelagos of the Azores and Madeira that includes 105,026 data entries between 1873 and 2021 (72% of the data occurring in 2000 and 2021). The methods used to collect the data were: live observations/captures (43%), sign surveys (35%), camera trapping (16%), bioacoustics surveys (4%) and radiotracking, and inquiries that represent less than 1% of the records. The data set includes 13 types of records: (1) burrows | soil mounds | tunnel, (2) capture, (3) colony, (4) dead animal | hair | skulls | jaws, (5) genetic confirmation, (6) inquiries, (7) observation of live animal (8), observation in shelters, (9) photo trapping | video, (10) predators diet | pellets | pine cones/nuts, (11) scat | track | ditch, (12) telemetry and (13) vocalization | echolocation. The spatial uncertainty of most records ranges between 0 and 100 m (76%). Rodentia (n =31,573) has the highest number of records followed by Chiroptera (n = 18,857), Carnivora (n = 18,594), Lagomorpha (n = 17,496), Cetartiodactyla (n = 11,568) and Eulipotyphla (n = 7008). The data set includes records of species classified by the IUCN as threatened (e.g., Oryctolagus cuniculus [n = 12,159], Monachus monachus [n = 1,512], and Lynx pardinus [n = 197]). We believe that this data set may stimulate the publication of other European countries data sets that would certainly contribute to ecology and conservation-related research, and therefore assisting on the development of more accurate and tailored conservation management strategies for each species. There are no copyright restrictions; please cite this data paper when the data are used in publications

Paraganglioma de grandes dimensões com invasão intracraniana

Head and neck paragangliomas usually present as a vascular, slow – growing and painless mass, localized in tympanic, jugular, vagal or common carotid artery area. The symptoms are variable and depend essentially on the affected site and the endocranial involvement. Case report: Male of 81 years-old, was send to our institution with a fever of unknown etiology, headache and asteny, with four weeks of evolution. The complementary study with CT, confirmed by MRI showed the presence of an expansive/infiltrative right jugulo-tympanic mass with intracranial extension, suggestive of a paraganglioma. Tumor was classified as class Di3 according to the Fisch’s modified classification. Other causes of fever were excluded, and paraneoplasic origin was assumed. Assessing the patient's age, the intracranial invasion and the unresectability of the tumor, the patient was proposed to start radiotherapy. Conclusions: Jugulo - tympanic paragangliomas are uncommon, requiring a high degree of clinical suspicion when the most frequent symptoms (pulsatile tinnitus and hearing loss) are absent. Although the primary therapeutic option is complete excision with preoperative embolization, in some cases this approach is not possible, and other clinical individual treatments should be considered.Os paragangliomas (PG) da cabeça e do pescoço apresentam-se habitualmente como massas vasculares, indolentes e não dolorosas, localizadas na região timpânica, jugular, vagal ou da artéria carótida comum. Os sintomas são variáveis e dependem essencialmente do local afetado. Caso Clínico: Homem de 81 anos, foi referenciado ao nosso hospital com febre de origem não determinada, cefaleias e astenia com quatro semanas de evolução. O estudo complementar com TC, e confirmado por RM, revelou a presença de uma massa expansiva/infiltrativa jugulo -timpânica direita com extensão intracraniana, sugestiva da presença de um paraganglioma. O tumor foi classificado como um tipo Di3, de acordo com a classificação modificada de Fisch. Outras causas para a febre foram excluídas, sendo assumida uma origen paraneoplásica. Tendo em conta a idade do doente, a invasão intracraniana e a irressecabilidade do tumor, o doente foi proposto para iniciar radioterapia. Conclusão: Os paragangliomas jugulo-timpânicos são incomuns, exigindo um elevado grau de suspeição clínica, principalmente quando os síntomas mais frequentes (acufenos pulsáteis e hipoacúsia) não estão presentes. Embora a opção terapéutica mais comumente utilizada seja a excisão completa com embolização pré-operatória, em alguns casos essa abordagem não é possível, sendo necessária uma avaliação individualizada, com ponderação de todas opções terapêuticas existente

Posterior orbital cellulitis as a complication of sphenoid sinusitis: a case report

Posterior orbital cellulitis is a clinical syndrome in which early severe visual loss dominates or precedes accompanying inflammatory orbital signs. This syndrome occurs as a complication of sphenoethmoiditis with involvement of the orbital apical segment of the optic nerve, and therefore it may be considered as a partial orbital apex syndrome. If a Posterior orbital cellulitis is suspected, an imaging diagnosis should be established and early treatment initiated to prevent irreversible blindness. We present a case of a 14-year-old patient, which fulfilled the criteria of a “posterior orbital cellulitis”A celulite orbitária posterior é uma síndrome clínica caracterizada por uma perda de visão severa que precede ou domina eventuais sinais inflamatórios orbitários. Esta síndrome ocorre como uma complicação de uma esfeno-etmoidite com envolvimento do segmento apical orbitário do nervo óptico, e portanto pode ser considerada como uma síndrome parcial do ápex orbitário. Se houver suspeita de uma celulite orbitária posterior, um diagnóstico de imagem deve ser estabelecido e deve ser iniciado tratamento precoce de forma a prevenir uma amaurose irreversível. Apresentamos um caso de uma doente de 14 anos, que cumpriu os critérios de uma "celulite orbitária posterior"

Cap. IV : Transformação digital e novas tecnologias militares

A Guerra ao Terror foi a agenda norte-americana que abriu um precedente para a forma como os reclusos da prisão de Guantánamo foram tratados, sendo considerados, sob a perspetiva dos EUA como “combatentes inimigos ilegais’’. O principal objetivo deste paper prende-se com uma análise da presença de terrorismo de estado na Prisão de Guantánamo, consistindo numa violação dos direitos humanos, sob a lente da Segurança Humana. Para esse fim, foi elaborada a seguinte pergunta de investigação que servirá de auxílio para conduzir a investigação: De que forma o terrorismo de Estado na prisão de Guantánamo constituiu uma violação dos direitos humanos? Foi ainda elaborada uma delimitação temporal com início em 2002, após a criação da prisão, e cujo término será em 2022 para que seja possível fazer um paralelo com a atualidade do status deste estabelecimento prisional.Na sequência da Resolução 1325 (2000) das Nações Unidas sobre Mulheres, Paz e Segurança (MPS), que apelou aos Estados para integrarem uma perspetiva de género nas suas políticas de segurança e construção da paz, a União Europeia (UE) tem dados alguns passos no sentido de integrar as questões de género na sua política externa e de segurança. Apesar destes desenvolvimentos empíricos, a literatura sobre segurança no espaço europeu não tem prestado a atenção devida aos processos através dos quais as questões de género e a agenda MPS podem contribuir para a construção da UE como um ator global. Este artigo procura aprofundar o debate sobre global actorness numa perspetiva de género, tendo em conta as seguintes questões: (1) que dimensões são relevantes para conceptualizar a UE como um actor global? (2) como é que a igualdade de género e a agenda MPS estão integradas na política externa e de segurança da UE? (3) como é que a UE moldou e promoveu a agenda MPS na arena global? Para responder a estas questões, partimos do modelo conceptual de global actorness de Bretherton e Vogler (2006) e na literatura sobre as dimensões de género da política externa e de segurança da União Europeia.Improvement of security governance on climate security is critical if fast-paced threats caused by the emission of greenhouse gases are to be dealt with. This article envisions climate security as a global public good, moving the debate beyond traditional security discussions and refocusing attention upon climate security governance for its effective provision. Additionally, having in mind the recognition granted to sub-state authorities under the Paris Agreement, it suggests that the provision of security governance in matters of climate change requires a multilevel governance (MLG) approach whereby sub-state actors become inescapable agents of climate security governance. In doing so, it is our wish to move the debate forward in two complementary ways: first, by expanding the notion of security in order to perceive climate security as a global public good and second, by shedding light on the reinforced role of sub-state authorities under the Paris agreement using the framework of MLGEste artigo analisa a noção de ataque constante do art.º 49.º do I PA às CG e a sua adequabilidade em relação às ciberoperações militares. A metodologia assentou na revisão do estado de arte, no recurso às leis e às regras jurídicas aplicáveis, à jurisprudência, à doutrina e à soft law. Foram analisadas as diferentes correntes doutrinárias (Schmitt, Dörmann e Melzer) e conclui-se que o conceito de hostilidade, embora não seja perfeito, todavia, é o que oferece mais garantias para proteger os civis, população civil e objetos civis.No mundo globalizado atual, o ciberespaço apresenta inúmeras oportunidades de cooperação e progresso, as também de ameaças e ataques. A cibersegurança dos Estados pode-se encontrar em risco, na falta de formação pertinente, recursos financeiros ou cooperação eficaz. O presente trabalho analisa de que maneira Portugal se encontra preparado para enfrentar as ameaças cibernéticas. Para isso, são apresentadas as principais tendências em cibercriminalidade em Portugal e os mecanismos de lidar com os desafios. Sugere-se que os ciberincidentes agravam as vulnerabilidades estatais, criam alguma desconfiança nas instituições nacionais. Porém, simultaneamente, impulsionam os avanços tecnológicos em ciberproteção. As capacidades nacionais estão, por isso, a ser direcionados para a formação tecnológica, a consolidação do corpo jurídico e o desenvolvimento de medidas de proteção, em parcerias regionais e internacionaisinfo:eu-repo/semantics/publishedVersio

A Case of XX Disorder of Sexual Development in a Female-Phenotype Roe Deer (<i>Capreolus capreolus</i> L.) Associated with Antlers Growth with Retained Velvet

A 3-to-4-year-old roe deer (Capreolus capreolus L.) was admitted to the Veterinary Hospital. Although it showed well-developed antlers with retained velvet, an external female appearance and genitalia were evident. External biometrical measurements were taken for the antlers, and a computed tomography was performed. Molecular studies targeting the SRY gene were performed, and a PIS (polled intersex syndrome) mutation diagnosis was implemented. The gonads consisted of a right testicle paired with a left ovotestis. Histologically, the ovary-like structures in the ovotestis were functional, but the testis, as the testis-like structure in the ovotestis, did not show active spermatogenesis. No evidence of SRY gene was detected by PCR, suggesting an XX-chromosome constitution. Additionally, polled intersex syndrome (PIS) deletion was not detected in the case under study. The clinical and histopathological findings confirmed the DSD with the presence of a testicle and a contralateral ovotestis