Lúpus Eritematoso: Manifestações Cutâneas e Tratamento

Cutaneous lupus erythematosus (CLE) includes a broad range of dermatologic manifestations, which may or may not be associated with systemic manifestations. Specific CLE - defined by the presence of an interface dermatitis on histopathological evaluation - is divided into several sub-types, namely acute CLE (ACLE), subacute CLE (SCLE) and chronic CLE (CCLE). CCLE includes discoid lupus erythematosus (DLE), as well as other rarer forms such as LE profundus (LEP). Nonspecific skin findings, such as livedo reticularis or purpura are more frequently seen in patients with systemic disease. Diagnosis requires classification of the subtype, through a combination of physical examination, laboratory studies, histology and sometimes direct immunofluorescence, at the same time ensuring to exclude systemic disease. Regarding the treatment of CLE, antimalarials and topical steroids continue to be the standard of care; however, immunosuppressants, thalidomide analogs and monoclonal antibodies are possible systemic therapies for recalcitrant disease. Patient education on proper sun protection and avoidance of triggers is crucial. This paper reviews the clinical manifestations of CLE, as well as the treatment.Lúpus eritematoso cutâneo engloba um vasto leque de manifestações dermatológicas, que podem ou não acompanhar- se de acometimento sistémico. As lesões cutâneas específicas – definidas histologicamente pela presença de dermatite de interface – são ainda dividas em subtipos, concretamente lúpus agudo, subagudo ou crónico. O subtipo crónico inclui ainda o lúpus discóide, bem como variantes mais raras, como o lúpus profundo. As manifestações cutâneas não específicas, nomeadamente o livedo reticular ou purpura, são mais frequentes nos doentes com acometimento sistémico. A abordagem diagnóstica implica a correcta subclassificação do subtipo, através duma combinação de exame físico, estudo laboratorial, análise histológica e ocasionalmente imunofluorescência directa, sendo imperativo a exclusão de atingimento sistémico. Do ponto de vista terapêutico, os corticosteróides tópicos e antimaláricos permanecem como a base da terapêutica; no entanto, imunossupressores, análogos da talidomida e anticorpos monoclonais são terapias sistémicas disponíveis para o tratamento da doença recalcitrante. A educação do doente acerca de medidas de fotoproteção e evicção de factores despoletantes é fulcral

Glycosaminoglycan Storage Disorders: A Review

Impaired degradation of glycosaminoglycans (GAGs) with consequent intralysosomal accumulation of undegraded products causes a group of lysosomal storage disorders known as mucopolysaccharidoses (MPSs). Characteristically, MPSs are recognized by increased excretion in urine of partially degraded GAGs which ultimately result in progressive cell, tissue, and organ dysfunction. There are eleven different enzymes involved in the stepwise degradation of GAGs. Deficiencies in each of those enzymes result in seven different MPSs, all sharing a series of clinical features, though in variable degrees. Usually MPS are characterized by a chronic and progressive course, with different degrees of severity. Typical symptoms include organomegaly, dysostosis multiplex, and coarse facies. Central nervous system, hearing, vision, and cardiovascular function may also be affected. Here, we provide an overview of the molecular basis, enzymatic defects, clinical manifestations, and diagnosis of each MPS, focusing also on the available animal models and describing potential perspectives of therapy for each one

Púrpura Fulminante numa Jovem de 20 Anos

A previously healthy 20-year-old female presented with extensive retiform purpura located at the face, upper and lower limbs, one week after an episode of acute tonsillitis. Despite the exuberance of the cutaneous findings and progression to skin necrosis she had no accompanying symptoms. Laboratory investigation revealed a heterozygous protein C mutation (exon 9, c.1332G> C, p.Trp444Cys), accounting for a partial deficiency of this anticoagulant protein. The patient was started on broad spectrum antibiotics, anticoagulation and systemic corticosteroids, with no lesional progression and complete resolution of cutaneous ulceration within 6 months. This is a singular case of purpura fulminans, since two different causative factors precipitated the events. The previous tonsillitis reported by the patient is significant, because the serum concentration of protein S may also decrease after an infectious event - post-infectious purpura fulminans. This case illustrates that purpura fulminans due to autoantibodies against protein S, although rare, should be considered, especially in the absence of a severe acute infection. It also illustrates how in a given patient different independent factors can act simultaneously, triggering potentially devastating clinical scenarios.Apresentamos o caso de uma jovem do sexo feminino, de 20 anos de idade, previamente saudável, que se apresentou com lesões muito extensas de púrpura retiforme localizadas na face, membros superiores e inferiores, uma semana após um episódio de amigdalite aguda. Apesar das lesões cutâneas exuberantes que evoluiram para necrose cutânea, a doente não apresentava qualquer sintomatologia associada. A investigação laboratorial revelou uma mutação em heterozigotia da proteína C (exon 9, c.1332G> C, p.Trp444Cys), condicionando um deficit parcial desta proteína anticoagulante. Iniciou antibioticoterapia de largo espectro, anticoagulação e corticoterapia sistémica, sem progressão das lesões no imediato, e resolução completa das ulcerações em 6 meses. Este é um caso singular de púrpura fulminante, visto que que dois fatores etiológicos diferentes precipitaram os eventos. A amigdalite prévia rela- tada pela doente é significativa, pois a concentração sérica da proteína S também pode diminuir após um evento infeccioso - purpura fulminans pós-infecciosa. Este caso ilustra que a púrpura fulminante decorrente de auto-anticorpos contra a proteína S, embora rara, deve ser considerada, principalmente na ausência de infecção aguda grave. Também evidencia como, em um determinado doente, diferentes fatores independentes podem atuar simultaneamente, desencadeando cenários clínicos potencialmente devastadores

Metabolic drivers of IC-BEVS productivity: Tackling the production of enveloped viral particles

The Insect Cell-Baculovirus Expression System (IC-BEVS) has a major track record for the production of recombinant proteins and vaccines. Although its widespread use, the physiological aspects that contribute to systems productivity are still to be fully disclosed. In the present work, the metabolic features of the two main insect host cell lines, Sf9 and High Five, were analyzed during cellular growth and after baculovirus infection for the production of enveloped Influenza VLPs (Inf-VLPs). The gathered data were contextualized in a metabolic network representative of central carbon and nitrogen metabolism. Metabolic Flux Analysis (MFA) was performed to have a quantitative overview of the cellular fluxome dynamics that followed infection. In addition, the main carbon sources that contributed most to flux activity were identified. The impact of baculovirus infection on the physiology of High Five and Sf9 host cell lines was assessed by metabolomics, aiming at the identification of metabolic markers of productivity. The information herein generated was used to design tailored supplementation schemes that could boost IC-BEVS production yields of two enveloped viral particles: influenza VLPs (Inf-VLP) as a vaccine candidate and the recombinant baculovirus (BV). The strong correlation observed between the metabolic state of the host cell and baculovirus infection highlights the capacity of this virus to act as a metabolic engineer, re-directing the cellular fluxome to support virus replication and production. The results also show that the viral load influence the cellular responsiveness to the supplements, with lower MOIs retrieving higher improvements in specific productivity. The careful selection of the MOI, along with the supplementation of culture medium with compounds altering cellular redox state and cholesterol metabolism, yielded a 6-fold improvement of specific productivity. These results pave the way to deepen our knowledge on the relationship between host cell and virus, contributing to the disclosure of the metabolic determinants that contribute to productivity

Less is More: an overview on the use of RNAi as a tool to achieve Substrate Reduction in Mucopolysaccharidoses

Lysosomal storage diseases (LSDs) are a group of genetic disorders caused by dysfunction in enzymes responsible for the intralysosomal degradation of particular compounds. Given their complex nature and the limitations of available therapies, the shift towards the development of combination treatments to counteract more effectively the pathological burden of these disorders is in the agenda of current research viewing to improve the clinical outcome of LSD patients. We consider that treatment strategies relying on RNA interference (RNAi), as well as in other RNA-based methodologies, may be feasible and particularly promising if designed in the context of a synergistic combinatorial therapeutic approach. We are currently evaluating an RNAi-dependent strategy based upon the selective downregulation of genes involved in the biosynthesis of glycosaminoglycans (GAGs), the major substrates that accumulate in patients suffering from a subset of LSDs called mucopolysaccharidoses (MPSs). Although enzyme replacement therapy is already available for some MPSs, it has some serious drawbacks, justifying the challenge of developing additional therapies targeting this group of disorders. Our goal is to promote an effective reduction of the accumulating substrate, ultimately decreasing or delaying MPSs’ symptoms. It should be noticed that, even though some substrate reduction therapy (SRT) drugs have already been approved (miglustat for GD) or are undergoing clinical trial (genistein and/or rhodamine B for MPSs), clinical evaluation of those same drugs has unveiled a few side effects, the most well-known being those observed for miglustat, which included osmotic diarrhea and weight loss. Nevertheless, chemical drugs aren’t the only way to achieve substrate reduction. Ours is fully molecular, drug-free approach, whose major focus relies on the biosynthetic pathways giving origin to each one of the GAGs whose degradation is impaired in MPS. Taking advantage of the RNAi technology potential, we have designed and assayed specific siRNAs targeting genes on those biosynthetic cascades to decrease the levels of production of each one of the four substrates. Their efficiency is currently being evaluated in vitro. Here we present an overview of the preliminary results of this project and unveil its next steps towards a full characterization/evaluation of its potential therapeutic effect.This work was partially supported by Fundação Millennium bcp (bcp/LIM/DGH/2014). Coutinho MF is grantee from the FCT (SFRH/BPD/101965/2014).N/

Coutinho et al. Less Is More: Substrate Reduction Therapy for Lysosomal Storage Disorders. Int. J. Mol. Sci. 2016, 17, 1065

Erratum for Less Is More: Substrate Reduction Therapy for Lysosomal Storage Disorders. [Int J Mol Sci. 2016]n/a.info:eu-repo/semantics/publishedVersio

Fronteiras na cidade de Tokio

Tese de Mestrado em Arquitectura com Especialização em Urbanismo Planeamento Urbano e Territorial