Transcriptional regulation of the mannosyltransferase-encoding gene pigm in inherited glycosylphosphatidylinositol (GPI) deficiency

RESUMO:O glicosilfosfatidilinositol (GPI) é um complexo glicolipídico utlizado por dezenas de proteínas, o qual medeia a sua ancoragem à superfície da célula. Proteínas de superfície celular ancoradas a GPI apresentam várias funções essenciais para a manutenção celular. A deficiência na síntese de GPI é o que caracteriza principalmente a deficiência hereditária em GPI, um grupo de doenças autossómicas raras que resultam de mutações nos genes PIGA, PIGL, PIGM, PIGV, PIGN, PIGO e PIGT, os quais sao indispensáveis para a biossíntese do GPI. Uma mutação pontual no motivo rico em GC -270 no promotor de PIGM impede a ligação do factor de transcrição (FT) Sp1 à sua sequência de reconhecimento, impondo a compactação da cromatina, associada à hipoacetilação de histonas, e consequentemente, impedindo a transcrição de PIGM. Desta forma, a adição da primeira manose ao GPI é comprometida, a síntese de GPI diminui assim como as proteínas ligadas a GPI à superficie das células. Pacientes com Deficiência Hereditária em GPI-associada a PIGM apresentam trombose e epilesia, e ausência de hemólise intravascular e anemia, sendo que estas duas últimas características definem a Hemoglobinúria Paroxística Nocturna (HPN), uma doença rara causada por mutações no gene PIGA. Embora a mutação que causa IGD seja constitutiva e esteja presente em todos os tecidos, o grau de deficiência em GPI varia entre células do mesmo tecido e entre células de tecidos diferentes. Por exemplo nos granulócitos e linfócitos B a deficiência em GPI é muito acentuada mas nos linfócitos T, fibroblastos, plaquetas e eritrócitos é aproximadamente normal, daí a ausência de hemólise intravascular. Os eventos transcricionais que estão na base da expressão diferencial da âncora GPI nas células hematopoiéticas são desconhecidos e constituem o objectivo geral desta tese. Em primeiro lugar, os resultados demonstraram que os níveis de PIGM mRNA variam entre células primárias hematopoiéticas normais. Adicionalmente, a configuração dos nucleossomas no promotor de PIGM é mais compacta em células B do que em células eritróides e tal está correlacionado com os níveis de expressão de PIGM, isto é, inferior nas células B. A presença de vários motivos de ligação para o FT específico da linhagem megacariocítica-eritróide GATA-1 no promotor de PIGM sugeriu que GATA-1 desempenha um papel regulador na sua transcrição. Os resultados mostraram que muito possivelmente GATA-1 desempenha um papel repressor em vez de activador da expressão de PIGM. Resultados preliminares sugerem que KLF1, um factor de transcrição restritamente eritróide, regula a transcrição de PIGM independentemente do motivo -270GC. Em segundo lugar, a investigação do papel dos FTs Sp demonstrou que Sp1 medeia directamente a transcrição de PIGM em ambas as células B e eritróide. Curiosamente, ao contrário do que acontece nas células B, em que a transcrição de PIGM requer a ligação do FT geral Sp1 ao motivo -270GC, nas células eritróides Sp1 regula a transcrição de PIGM ao ligar-se a montante e não ao motivo -270GC. Para além disso, demonstrou-se que Sp2 não é um regulador directo da transcrição de PIGM quer nas células B quer nas células eritróides. Estes resultados explicam a ausência de hemólise intravascular nos doentes com IGD associada a PIGM, uma das principais características que define a HPN. Por último, resultados preliminares mostraram que a repressão da transcrição de PIGM devida à mutação patogénica -270C>G está associada com a diminuição da frequência de interacções genómicas em cis entre PIGM e os seus genes “vizinhos”, sugerindo adicionalmente que a regulação de PIGM e desses genes é partilhada. No seu conjunto, os resultados apresentados nesta tese contribuem para o conhecimento do controlo transcricional de um gene housekeeping, específico-detecido, por meio de FTs genéricos e específicos de linhagem.-------------ABSTRACTC: Glycosylphosphatidylinositol (GPI) is a complex glycolipid used by dozens of proteins for cell surface anchoring. GPI-anchored proteins have various functions that are essential for the cellular maintenance. Defective GPI biosynthesis is the hallmark of inherited GPI deficiency (IGD), a group of rare autosomal diseases caused by mutations in PIGA, PIGL, PIGM, PIGV, PIGN, PIGO and PIGT, all genes indispensable for GPI biosynthesis. A point mutation in the -270GC-rich box in the core promoter of PIGM disrupts binding of the transcription factor (TF) Sp1 to it, imposing nucleosome compaction associated with histone hypoacetylation, thus abrogating transcription of PIGM. As a consequence of PIGM transcriptional repression, addition of the first mannose residue onto the GPI core and thus GPI production are impaired; and expression of GPI-anchored proteins on the surface of cells is severely impaired. Patients with PIGM-associated IGD suffer from life-threatening thrombosis and epilepsy but not intravascular haemolysis and anaemia, two defining features of paroxysmal nocturnal haemoglobinuria (PNH), a rare disease caused by somatic mutations in PIGA. Although the disease-causing mutation in IGD is constitutional and present in all tissues, the degree of GPI deficiency is variable and differs between cells of the same and of different tissues. Accordingly, GPI deficiency is severe in granulocytes and B cells but mild in T cells, fibroblasts, platelets and erythrocytes, hence the lack of intravascular haemolysis.The transcriptional events underlying differential expression of GPI in the haematopoietic cells of PIG-M-associated IGD are not known and constitute the general aim of this thesis. Firstly, I found that PIGM mRNA levels are variable amongst normal primary haematopoietic cells. In addition, the nucleosome configuration in the promoter of PIGM is more compacted in B cells than in erythroid cells and this correlated with the levels of PIGM mRNA expression, i.e., lower in B cells. The presence of several binding sites for GATA-1, a mega-erythroid lineage-specific transcription factor (TF), at the PIGM promoter suggested that GATA-1 has a role on PIGM transcription. My results showed that GATA-1 in erythroid cells is most likely a repressor rather than an activator of PIGM expression. Preliminary data suggested that KLF1, an erythroid-specific TF, regulates PIGM transcription but independently of the -270GC motif. Secondly, investigation of the role of the Sp TFs showed that Sp1 directly mediates PIGM transcriptional regulation in both B and erythroid cells. However, unlike in B cells in which active PIGM transcription requires binding of the generic TF Sp1 to the -270GC-rich box, in erythroid cells, Sp1 regulates PIGM transcription by binding upstream of but not to the -270GC-rich motif. Additionally, I showed that Sp2 is not a direct regulator of PIGM transcription in B and erythroid cells. These findings explain lack of intravascular haemolysis in PIGM-associated IGD, a defining feature of PNH. Lastly, preliminary work shows that transcriptional repression of PIG-M by the pathogenic -270C>G mutation is associated with reduced frequency of in cis genomic interactions between PIGM and its neighbouring genes, suggesting a shared regulatory link between these genes and PIGM. Altogether, the results presented in this thesis provide novel insights into tissuespecific transcriptional control of a housekeeping gene by lineage-specific and generic TFs.

O Conselho de Estado na monarquia constitucional: uma reflexão preliminar

This article summarises the transformations in the State Council’s functions and membership throughout the Portuguese Constitutional Monarchy, and makes a preliminary attempt to scrutinise the political role played by an institution designed since its inception to advise the monarch. In spite of the parsimony of contemporary sources, and even contradictory empirical evidence, it seems indisputable that in several critical occasions the monarch’s political decisions were influenced by the dominant view in the State Council. Finally, the article presents the collective biography of the 73 individuals appointed to the State Council between 1833 and 1910 – who may be defined as the inner circle of the ruling elite - focusing on basic background features (birthplace, age, education, occupation, noble titles and political experience).

Liberdade e tolerância religiosa: "portugueses não católicos" no ultramar do século XIX

The personal legal status of native non catholic people of Portuguese colonial Empire during the nineteenth century, their position in what concerns Portuguese citizenship, is the main subject of this article. While discussing constitutional articles on religion, Portuguese deputies of the nineteenth century were confronted with a set of problems about that status which they find difficult to solve: should non catholic peoples who were born in Portuguese colonial territory be treated as plain Portuguese citizens or where they just “savage people”, “colonial subjects” or, in a more optimistic approach, “civilizing subjects”. The results were not conclusive, giving rise to an “uncertainty principle” which enabled central and local government to decide in an almost casuistic way about native people status and rights.

A Constituição e o Direito Justo

Também publicado em DELGADO, José Pina (org.) e SILVA, Mário Ramos Pereira (org.) "Estudos em Comemoração do XXº Aniversário da Constituição da República de Cabo Verde", Praia: Edições do ISCJS, 2013, [9]-29 pp

A restrição ao sigilo bancário decorrente do regime de branqueamento de capitais

Technologic and socio-economic mutations have always determined challenges not only to lawyers, but to law itself. These phenomena have occurred specially when trying to deal with the hard task of finding solutions for the current increasing mismatch of social interests, for example, bank secrecy and money laundering. Usually occurring simultaneously, they are typical examples of outcomes generated by technological and socio-economic innovations that have become fashionable and captured international attention. At the same time, bank secrecy and money laundering support interests belonging to different dimensions, deserving to be balanced in the light of the heterogeneous mechanisms provided by the law to its practitioners and society as a hole. In order to achieve an outcome in accordance with the Rule of Law´s principles, lawyers´ tools are consequently subordinated to constitutional and social justice. Guided by this purpose, we performed the present study, aiming to analyse bank secrecy and money laundering in the light of the current stablished juridical procedures. We intended to develop a prudent point of view that is also in accordance with social reality. In sum, we demonstrate that bank secrecy should adopt a flexible character, embedding new settings and following the socio-economic path in a globalized world with constant innovations.

A intervenção dos orgãos de polícia criminal em sede de julgamento

Currently, Portugal assumes itself as a democratic rule of substantive law State, sustained by a legal system seeking the right balance between the guarantee of fundamental rights and freedoms constitutional foreseen in Portugal’s Fundamental Law and criminal persecution. The architecture of the penal code lies with, roughly speaking, a accusatory basic structure, “deliberately attached to one of the most remarkable achievements of the civilizational democratic progress, and by obedience to the constitutional commandment”, in balance with the official investigation principle, valid both for the purpose of prosecution and trial. Regarding the principle of non self-incrimination - nemo tenetur se ipsum accusare, briefly defined as the defendant’s right of not being obliged to contribute to the self-incrimination, it should be stressed that there isn’t an explicit consecration in the Portuguese Constitution, being commonly accepted in an implicit constitutional prediction and deriving from other constitutional rights and principles, first and foremost, the meaning and scope of the concept of democratic rule of Law State, embedded in the Fundamental Law, and in the guidelines of the constitutional principles of human person dignity, freedom of action and the presumption of innocence. In any case, about the (in) applicability of the principle of the prohibition of self-incrimination to the Criminal Police Bodies in the trial hearing in Court, and sharing an idea of Guedes Valente, the truth is that the exercise of criminal action must tread a transparent path and non-compliant with methods to obtain evidence that violate the law, the public order or in violation of democratic principles and loyalty (Guedes Valente, 2013, p. 484). Within the framework of the penal process relating to the trial, which is assumed as the true phase of the process, the witness represents a relevant figure for the administration of criminal justice, for the testimonial proof is, in the idea of Othmar Jauernig, the worst proof of evidence, but also being the most frequent (Jauernig, 1998, p. 289). As coadjutant of the Public Prosecutor and, in specific cases, the investigating judge, the Criminal Police Bodies are invested with high responsibility, being "the arms and eyes of Judicial Authorities in pursuing the criminal investigation..." which has as ultimate goal the fulfillment of the Law pursuing the defense of society" (Guedes Valente, 2013, p. 485). It is in this context and as a witness that, throughout operational career, the Criminal Police Bodies are required to be at the trial hearing and clarify the Court with its view about the facts relating to occurrences of criminal context, thus contributing very significantly and, in some cases, decisively for the proper administration of the portuguese criminal justice. With regards to the intervention of Criminal Police Bodies in the trial hearing in Court, it’s important that they pay attention to a set of standards concerning the preparation of the testimony, the very provision of the testimony and, also, to its conclusion. Be emphasized that these guidelines may become crucial for the quality of the police testimony at the trial hearing, thus leading to an improvement of the enforcement of justice system. In this vein, while preparing the testimony, the Criminal Police Bodies must present itself in court with proper clothing, to read before and carefully the case files, to debate the facts being judged with other Criminal Police Bodies and prepare potential questions. Later, while giving his testimony during the trial, the Criminal Police Bodies must, summing up, to take the oath in a convincing manner, to feel comfortable, to start well by convincingly answering the first question, keep an attitude of serenity, to adopt an attitude of collaboration, to avoid the reading of documents, to demonstrate deference and seriousness before the judicial operators, to use simple and objective language, to adopt a fluent speech, to use nonverbal language correctly, to avoid spontaneity responding only to what is asked, to report only the truth, to avoid hesitations and contradictions, to be impartial and to maintain eye contact with the judge. Finally, at the conclusion of the testimony, the Criminal Police Bodies should rise in a smooth manner, avoiding to show relief, resentment or satisfaction, leaving a credible and professional image and, without much formality, requesting the judge permission to leave the courtroom. As final note, it’s important to stress that "The intervention of the Police Criminal Bodies in the trial hearing in Court” encloses itself on a theme of crucial importance not only for members of the Police and Security Forces, who must welcome this subject with the utmost seriousness and professionalism, but also for the proper administration of the criminal justice system in Portugal.

Cumprimento de ordens, obediência hierárquica e disciplina militar versus perpetração (in)voluntária de crimes

Following orders, hierarchical obedience and military discipline are essential values for the survival of the armed forces. Without them, it is not possible to conceive the armed forces as an essential pillar of a democratic state of law and a guarantor of national independence. As issuing orders as well as receiving and following them are inextricably linked to military discipline, and as such injunctions entail the workings of a particular obedience regime within the specific kind of organized power framework which is the Armed Forces, only by analysing the importance of such orders within this microcosm – with its strict hierarchical structure – will it be possible to understand which criminal judicial qualification to ascribe to the individual at the rear by reference to the role of the front line individual (i.e. the one who issues an order vs the one who executes it). That is, of course, when we are faced with the practice of unlawful acts, keeping in mind the organizational framework and its influence over the will of the executor. One thing we take as read, if the orders can be described as unlawful, the boundary line of the duty of obedience, which cannot be overstepped, both because of a legal as well as a constitutional imperative, will have been crossed. And the military have sworn an oath of obedience to the fundamental law. The topic of hierarchical obedience cannot be separated from the analysis of current legislation which pertains to the topic within military institutions. With that in mind, it appeared relevant to address the major norms which regulate the matter within the Portuguese military legal system, and, whenever necessary and required by the reality under analysis, to relate that to civilian law or legal doctrine.

A equidade na indeminização dos danos não patrimoniais

This study examines the quantification of compensation for non-pecuniary damage, awarded by means of judicial decisions based on equity, and seeks to verify whether such calculation safeguards legal certainty and predictability when applying the law, as well as whether it observes the principles of proportionality and equality. Firstly, the limits for discretionary judgment permitted to the judge were determined, by evaluating the criteria established under the law. Then, by examining the grounds of the judicial decisions in cases that had been selected beforehand, this study sought to detect operation modes in concrete considerations of equity used by judges. The examination of the grounds on which these judicial decisions are based permitted the comprehension of the calculation method used in each case and the observation that the criteria of compensatory nature, such as the extent of the damage and the respective consequences, assumed a primary role. Despite discrepancies in viewpoints with regard to certain issues of law, the jurisprudence examined reveals that great care is taken to consider the solutions reached in similar cases, in an attempt to ensure that the different criteria applied in the quantification of compensation are given uniform relevance. The comparison of decisions, reported to cases with similar legal contours, did not reveal relevant discrepancies in the calculation criteria used, nor are they disproportionate regarding the amount of compensation awarded, which means that resorting to equity, in determining the compensation to be awarded due to nonpecuniary damage, does not jeopardize legal certainty or predictability when applying the law, and observes the principle of proportionality, which is anchored in the constitutional principle of equality. The study performed, led to the conclusion that the grounds on which judicial decisions are based, by itemising the elements which are taken into account and the criteria adopted by the judge, allow these to be taken into consideration in similar cases, contributing towards uniform interpretation and application of the law, ensuring legal certainty and predictability when resorting to equity while quantifying compensation.

A consagração dos naturais

“One cannot analyse a legal concept outside the economic and socio-cultural context in which it was applied” – such is the longstanding thesis of António Manuel Hespanha. I argue that Hespanha’s line of argument relative to legal concepts is also applicable, mutatis mutandis, to legal agents: the magistrates, advocates, notaries, solicitors and clerks who lived and exercised their professions in a given time and place. The question, then, is how to understand the actions of these individuals in particular contexts – more specifically in late 18th century and 19th century Goa. The main goal of the present thesis was to comprehend how westernized and Catholic Goan elite of Brahman and Chardó origin who provided the majority of Goan legal agents used Portuguese law to their own advantage. It can be divided into five key points. The first one is the importance of the Constitutional liberalism regime (with all the juridical, judicial, administrative and political changes that it has brought, namely the parliamentary representation) and its relations with the perismo – a local political and ideological tendency nurtured by Goan native Catholic elite. It was explored in the chapter 2 of this thesis. The second key point is the repeated attempts made by Goan native Catholic elite to implement the jury system in local courts. It was studied in the chapter 3. Chapter 4 aims to understand the participation of the native Catholic elite in the codification process of the uses and traditions of the indigenous peoples in New Conquests territory. The fourth key point is the involvement of those elites not only in the conflict of civil and ecclesiastical jurisdictions but also in the succession of the Royal House of Sunda. It was analyzed in the chapter 5. The functions of an advocate could be delegated to someone who, though lacking a law degree, possessed sufficient knowledge to perform this role satisfactorily. Those who held a special licence to practice law were known as provisionários (from provisão, or licence, as opposed to the letrados, or lettered). In the Goa of the second half of the 18th century and the 19th century, such provisionários were abundant, the vast majority coming from the native Catholic elite. The characteristics of those provisionários, the role played by the Portuguese letrados in Goa and the difficult relations between both groups were studied in the chapter 6.

O sigilo bancário e a fraude fiscal

Tax evasion and fraud threaten the economic and social objectives of modern tax systems, precluding the state funding for the satisfaction of collective needs and the fair distribution of wealth, being a violation of basic principles and values of our society. In tax law, to give tax administration the necessary powers to supervise and control the information provided by taxpayers and combat tax evasion and fraud, over the last years the grounds for a derogation of bank secrecy without judicial authorization have been extended, which raises some constitutional compatibility issues. Similarly, this tendency of making this legal regime more flexible and increasing automatic exchange of information has been followed by the European Union and the international community. Banking secrecy, as a professional secrecy, is an instrument to protect the right to privacy but also appears as an anti-abuse and repressive mechanism of evasive and fraudulent behaviors. Because of the conflict of interests will always be necessary to make a practical agreement between them, ensuring the legality and the due guarantees of the taxpayers but also an effective way to combat tax evasion and fraud. Bank secrecy cannot be one method to, behind the right to privacy, taxpayers practice illegal activities. But the practice of these irregular conducts also does not justify a total annihilation of the right to banking secrecy, uncovering all documents and bank information’s. Although considering the legislative changes, the administrative derogation of bank secrecy will always be what the tax administration does of it.

Da intercepção de comunicações, acções encobertas e serviços de informações

The Portuguese Intelligence Services have their operational skills limited due to the grievances caused by the Dictatorship and, in particular, by its political police. With the help of historical elements, and by analyzing current legislation, we demonstrate that such grievances are today unjustified and misplaced, mainly taking into account the Risk Society’s multifaceted threats. Also part of our analysis is the impugnment of the Constitutional Court’s decision nº 413/2015, which pronounced unconstitutional the norm contained in Decree nº 426/XII, of the Republic’s Assembly, article nº 78, nº2, which intended to allow Intelligence Services access to the so-called “metadata”, as well as to tax and banking information. It is our understanding, and we demonstrate it in our dissertation, that should be allowed the access of, not only the above mentioned information, but also the means known as communications interception and undercover operations to the Intelligence Services, as long as properly supervised and inspected.

O processo sumário e a reforma do Código do Processo Penal de 2013

The present work aims to develop the theme "The summary procedure and the reform of 2013". The purpose of its analysis serves the interest to understand the virtues and disadvantages of the changes introduced by Act n.º 20/2013 to our Code of Criminal Procedure, and the main focus of the present reflection is to further the impact of the measures taken by the legislator to the summary proceedings. The opening of the most serious crimes to summary procedure is a reform measure duly highlighted because it is a true innovation in the Portuguese penal system. Therefore, it urges to analyse not only the consequences of this measure, as well as if the objectives of its introduction in the summary procedure system are met. It should be noted that the legislator intends to promote speedy trial, and at the same time, ensure compliance with the Constitutional rights associated to the accused. At this point it is important to realize if there is a restriction of the accused essential guarantees. On the other hand, it should be noted that the typical characteristics of summary proceedings might have been invariably modified, due to the innovative aspect of the reform. That said, the changes might have fostered a mischaracterization of the typical format of the summary procedure, both in terms of the nature of the proceedings and in terms of its space and objectives within the penal system. Reflecting on the above will provide a deeper understanding of the volatile balance between the Portuguese governing prosecution efficiency and the Constitution, as well as the future of the criminal policy in Portugal.

Do Comandante Supremo das Forças Armadas

The President of the Portuguese Republic’s relation with the Armed Forces in the current political system is not confined to the vast legislative and constitutional set. As Chief of State and Supreme Commander of the Armed Forces, the President holds different powers. We develop an approach through the main concepts and themes that regulate the Armed Forces and National Defense, focusing the relations that involve the President directly. The attribution by inherence of the Supreme Command of the Armed Forces to the Chief of State, imposes a peculiar relation with the remainder sovereign institutions. The need to analyze the intermediate services as far as military and defense issues are concerned becomes necessary, in particular the role of the Military Cabinet – as a structure of direct support – and the High Council of National Defense – as an advisory body.

O regime penal das imunidades dos titulares dos órgãos políticos de soberania

Portugal is one of the countries that has a constitutional regime of immunity. This protects certain individuals in political positions from prosecution under the law. These individuals are said to have a privileged status when compared to ordinary citizens. The purpose of this study is to examine the immunities enjoyed by President, the members of Parliament and the government ministers. The regime of immunities can be found to generate a certain sense of injustice and feelings of mistrust since the individual can not, albeit temporarily, be held criminally responsible for criminal conduct. It is urgent, therefore, to find a consistent justification with the principles and values of the Criminal Law. The Parliament is the place of the exercise of democratic power and, therefore, a member of Parliament assumes a central position in parliamentary activity. For this reason, it will be necessary to determine analysis to determine the meaning the prerogative of criminal irresponsibility. One question that must be asked is to know how the dogmatic plan that the immunities of the political organs of sovereignty must be seen.