Lipid Nanoparticles as Carriers for RNAi against Viral Infections: Current Status and Future Perspectives

The efforts made to develop RNAi-based therapies have led to productive research in the field of infections in humans, such as hepatitis C virus (HCV), hepatitis B virus (HBV), human immunodeficiency virus (HIV), human cytomegalovirus (HCMV), herpetic keratitis, human papillomavirus, or influenza virus. Naked RNAi molecules are rapidly digested by nucleases in the serum, and due to their negative surface charge, entry into the cell cytoplasm is also hampered, which makes necessary the use of delivery systems to exploit the full potential of RNAi therapeutics. Lipid nanoparticles (LNP) represent one of the most widely used delivery systems for in vivo application of RNAi due to their relative safety and simplicity of production, joint with the enhanced payload and protection of encapsulated RNAs. Moreover, LNP may be functionalized to reach target cells, and they may be used to combine RNAi molecules with conventional drug substances to reduce resistance or improve efficiency. This review features the current application of LNP in RNAi mediated therapy against viral infections and aims to explore possible future lines of action in this field.

Development of nucleic acid vaccines: use of self-amplifying RNA in lipid nanoparticles

Self-amplifying RNA or RNA replicon is a form of nucleic acid-based vaccine derived from either positive-strand or negative-strand RNA viruses. The gene sequences encoding structural proteins in these RNA viruses are replaced by mRNA encoding antigens of interest as well as by RNA polymerase for replication and transcription. This kind of vaccine has been successfully assayed with many different antigens as vaccines candidates, and has been shown to be potent in several animal species, including mice, nonhuman primates, and humans. A key challenge to realizing the broad potential of self-amplifying vaccines is the need for safe and effective delivery methods. Ideally, an RNA nanocarrier should provide protection from blood nucleases and extended blood circulation, which ultimately would increase the possibility of reaching the target tissue. The delivery system must then be internalized by the target cell and, upon receptor-mediated endocytosis, must be able to escape from the endosomal compartment into the cell cytoplasm, where the RNA machinery is located, while avoiding degradation by lysosomal enzymes. Further, delivery systems for systemic administration ought to be well tolerated upon administration. They should be safe, enabling the multiadministration treatment modalities required for improved clinical outcomes and, from a developmental point of view, production of large batches with reproducible specifications is also desirable. In this review, the concept of self-amplifying RNA vaccines and the most promising lipid-based delivery systems are discussed.

Halocinese e tectônica de rafts na bacia emersa de Benguela (Angola): dados de campo, modelagem física e restauração estrutural

Este trabalho foi feito com dados de campo da Bacia de Benguela, na costa sul de Angola. Nesta área aflora uma plataforma carbonática de idade albiana, análoga às formações produtoras de petróleo de mesma idade na Bacia de Campos. Duas unidades principais compõem a seção albiana na área: uma unidade inferior caracterizada por carbonatos de águas rasas com intercalações de conglomerados e arenitos na base, totalizando 300 m de espessura total, e uma unidade superior formada por sedimentos carbonáticos de águas mais profundas com espessura de 100 m. Esta seção se apresenta em dois domínios estruturais. O norte é caracterizado por uma plataforma de cerca de 5 km de largura, 35 km de comprimento na direção NE e que se encontra próxima ao embasamento. No domínio sul, essa plataforma se apresenta em fragmentos de menos de 10 km de comprimento por até 5 km de largura, alguns deles afastados do embasamento. Dado que há evaporitos sob essa plataforma, fez-se este trabalho com o objetivo de se avaliar a influência da tectônica salífera na estruturação das rochas albianas. A utilização de imagens de satélite de domínio público permitiu a extrapolação dos dados pontuais e a construção de perfis topográficos que possibilitaram a montagem das seções geológicas. Os dados de campo indicam que a estruturação na parte norte, onde há menos sal, é influenciada pelas estruturas locais do embasamento. Já na parte sul, onde a espessura de sal é maior, a deformação é descolada das estruturas do embasamento. Foi feita uma seção geológica em cada domínio e a restauração dessas duas seções mostrou que a unidade albiana na parte sul sofreu uma distensão de 1,25 (125%) e a de norte se distendeu em 1,05 (105%). Essa diferença foi causada por uma maior espessura original de sal na parte sul, que fez com que a cobertura albiana se distendesse mais em consequência da fluência do sal mergulho abaixo, em resposta ao basculamento da bacia em direção a offshore. A intensidade da deformação na parte sul foi tal que os blocos falhados perderam os contatos entre si, formando rafts. Este efeito foi reproduzido em modelos físicos, indicando que a variação na espessura original de sal tem grande influência na estruturação das rochas que o cobrem. As variações de espessura de sal são controladas por zonas de transferência do embasamento e estas mesmas estruturas controlam os limites dos blocos que sofrem soerguimento diferencial no Cenozoico.

A study on the lipid distribution pattern in muscles of two teleosts, Arius dussumieri and Ophiocephalus striatus

The muscles of the various regions and zones of the body of the two teleosts, A. dussumieri and O. striatus have been analyzed for lipid contents. There is a significant dorsum-ventral gradient in lipid concentration exhibited by both the fishes with higher lipid values in the ventral aspect of the body, especially the belly flaps. As regards the vertical series, both the fishes exhibit comparatively higher lipid contents at the dorsal aspects of the caudal region and at anterior portion of the dorsal fin area with lower lipid values at cephalic and middle portions of the body. The red muscle of Arius exhibits higher lipid content than the white muscle.

Analysis of lipid and lipid-fractions of some freshwater fishes and their inter-relationship

Two fish species each from carnivorous (Clarias batrachus, Channa punctatus), omnivorous (Cyprinus carpio, Cirrhinus reba), and plankton feeder (Catla catla, Labeo rohita) were collected from freshwater sources under natural habitat to study their total lipid (TL) and lipid-fractions. Significant relationship between these parameters was also worked out. The variation of total lipid and lipid-fractions in tissues of freshwater fishes were not significantly different (P>0.05). But a higher trend of total lipid and glyceride (TGL) contents were found in carnivores followed by omnivores and least in plankton feeders. The trend was reverse for total phospholipid, cholesterol and free fatty acids. TGL content in all class of fishes was significantly related with TL (P<0.01), phospholipid (PL) (P<0.001), cholesterol (P<0.05), free fatty acids (P<0.05) and monoglycerides (P<0.001). Similarly total lipid was linearly related with total glycerides (TL=-3.02 + 0.10 TGL) and phospholipid (TL=7.13-0.12 PL). From this study it is concluded that almost all lipid-fractions of freshwater fishes can be predicted easily from total lipid content of the tissue.

Short-term preservation of Brachionus calyciflorus: effects of storage on protein, lipid and carbohydrate contents

The short-term preservation of Brachionus calyciflorus for 45 days at three different temperatures (4, -4 and -20°C) led to decrease in protein, lipid and carbohydrate contents in all the three cases. However, the rate of deterioration was much higher at 4C than at -4 and -20°C. At 4C, protein, lipid and carbohydrate contents reduced by 76.78, 81.11 and 62.83%, respectively, and at -4°C, these were 27.94, 37.46 and 18.42%, respectively, whereas at -20°C, the deterioration was limited to 9.28, 16.44 and 11.35%, respectively, when compared with the control values. Thus, preservation at -20°C is comparatively better as it exerts limited effect on the protein, lipid and carbohydrate contents of B. calycijlorus.