Improving the downstream processing of interferon alfa-2b using alternative purification platforms based on ionic liquids

Improvements on human life expectancy and the lack of effective therapies has led to an increment of chronic diseases, being the application of biopharmaceuticals an efficient strategy to mitigate this scenario. Among the current available biopharmaceuticals, the role of interferon α-2b (IFNα-2b) should be highlighted, as it has been marketed over 30 years with a considerable impact on the global therapeutic proteins market (Castro et al, Vaccines, 2021). IFN manufacturing requires the use of the recombinant DNA technology, involving two main stages, the upstream and downstream stages. The first includes recombinant protein production in a suitable host microorganism, such as Escherichia coli (Castro et al, Sep. Purif. Technol., 2020), while the second comprises protein recovery, isolation, purification and polishing. Due to the high demands of the pharmaceutical industry for products with high purity and biological activity, the downstream stage is responsible for the majority of the production costs of biopharmaceuticals (50–90%), often including time-consuming and multi-step processes. Therefore, there is an immediate need to develop more efficient, cost-effective, and sustainable protein purification methodologies. In this work, two ionic-liquid-(IL)-based strategies were investigated for the purification of IFNα-2b recombinantly produced from E. coli fermentation broth, namely as adjuvants in aqueous biphasic systems or as chromatographic ligands immobilized in solid materials. Overall, the obtained results demonstrate that by tailoring IL’s chemical structures, improved protein purification processes are obtained and that the secondary structure of proteins is preserved.publishe

Sustainable ionic-liquid-based strategies for the downstream processing of interferon α-2b from Echerichia coli

Over the last decades, society has been facing an increment of chronic diseases due to the higher human life expectancy and the lack of efficient treatments for several pathologies. In this regard, biopharmaceuticals have become one of the most effective clinical treatments for a broad range of diseases, including cancer, metabolic and neurodegenerative disorders [1]. Among biopharmaceuticals, the role of interferons, particularly interferon α-2b (IFNα-2b), should be underlined, as they have been marketed for over 30 years with a considerable impact on the global therapeutic proteins market [2]. Usually based on the recombinant DNA technology, the manufacturing process of biopharmaceuticals encompasses two main stages: the upstream and downstream stages. Typically, the upstream phase includes recombinant protein production processes in a suitable host microorganism, such as Escherichia coli [3], while the general downstream processing of biopharmaceuticals comprises four stages - recovery, isolation, purification and polishing -, which are responsible for the majority of the production costs of biopharmaceuticals (50–90%) [3]. The downstream processing is a time-consuming and multi-step process, for which the development of cost-effective purification processes is mandatory to decrease their costs and environmental impact. In this context, two ionic-liquid-(IL)-based strategies were investigated in this work for the purification of IFNα-2b recombinantly produced from E. coli fermentation broth. ILs have been used as adjuvants in aqueous two-phase systems (ATPS) and applied in supported materials as alternative ligands. The obtained results demonstrate that ILs have a tailoring ability and contribute to the development of more effective and sustainable downstream processes of biopharmaceuticals.publishe

Purification of interferon alpha 2b-based biopharmaceuticals using ionic liquid-based technologies

Protein biopharmaceuticals, among which interferon alpha-2b (IFNα-2b) that can be used in the treatment of chronic hepatitis C, have become an indispensable product of current medicine. Aiming at finding new cost-effective, efficient and sustainable technologies for recombinant IFNα-2b purification, ionic liquids were investigated as adjuvants in polymer-polymer aqueous two-phase systems (ATPS) or as chromatographic ligands covalently attached in silica (Supported ionic liquids, SILs). The application of ionic liquids as adjuvants in ATPS composed of polyethylene glycol (PEG 600 g/mol) and polypropylene glycol (PPG 400 g/mol) enhanced the purity of IFNα-2b recovered in the PEG-rich phase. On the other hand, SILs exhibited promising results toward IFNα-2b purification both in conditions favouring ionic or hydrophobic interactions, reinforcing the multimodal character of these novel stationary phases. Also, the secondary structure of IFNα-2b is preserved with both purification processes, as appraised by circular dichroism and western-blot studies. Overall, our results demonstrate the high potential exhibited by ionic liquids toward the preparative purification of the recombinant IFNα-2b biopharmaceuticals.publishe

Dengue Infection Increases the Locomotor Activity of Aedes aegypti Females

BACKGROUND: Aedes aegypti is the main vector of the virus causing Dengue fever, a disease that has increased dramatically in importance in recent decades, affecting many tropical and sub-tropical areas of the globe. It is known that viruses and other parasites can potentially alter vector behavior. We investigated whether infection with Dengue virus modifies the behavior of Aedes aegypti females with respect to their activity level. METHODS/PRINCIPAL FINDINGS: We carried out intrathoracic Dengue 2 virus (DENV-2) infections in Aedes aegypti females and recorded their locomotor activity behavior. We observed an increase of up to ∼50% in the activity of infected mosquitoes compared to the uninfected controls. CONCLUSIONS: Dengue infection alters mosquito locomotor activity behavior. We speculate that the higher levels of activity observed in infected Aedes aegypti females might involve the circadian clock. Further studies are needed to assess whether this behavioral change could have implications for the dynamics of Dengue virus transmission

Epidemiology of Disappearing Plasmodium vivax Malaria: A Case Study in Rural Amazonia

Background: New frontier settlements across the Amazon Basin pose a major challenge for malaria elimination in Brazil. Here we describe the epidemiology of malaria during the early phases of occupation of farming settlements in Remansinho area, Brazilian Amazonia. We examine the relative contribution of low-density and asymptomatic parasitemias to the overall Plasmodium vivax burden over a period of declining transmission and discuss potential hurdles for malaria elimination in Remansinho and similar settings. Methods: Eight community-wide cross-sectional surveys, involving 584 subjects, were carried out in Remansinho over 3 years and complemented by active and passive surveillance of febrile illnesses between the surveys. We used quantitative PCR to detect low-density asexual parasitemias and gametocytemias missed by conventional microscopy. Mixed-effects multiple logistic regression models were used to characterize independent risk factors for P. vivax infection and disease. Principal Findings/Conclusions P. vivax prevalence decreased from 23.8% (March–April 2010) to 3.0% (April–May 2013), with no P. falciparum infections diagnosed after March–April 2011. Although migrants from malaria-free areas were at increased risk of malaria, their odds of having P. vivax infection and disease decreased by 2–3% with each year of residence in Amazonia. Several findings indicate that low-density and asymptomatic P. vivax parasitemias may complicate residual malaria elimination in Remansinho: (a) the proportion of subpatent infections (i.e. missed by microscopy) increased from 43.8% to 73.1% as P. vivax transmission declined; (b) most (56.6%) P. vivax infections were asymptomatic and 32.8% of them were both subpatent and asymptomatic; (c) asymptomatic parasite carriers accounted for 54.4% of the total P. vivax biomass in the host population; (d) over 90% subpatent and asymptomatic P. vivax had PCR-detectable gametocytemias; and (e) few (17.0%) asymptomatic and subpatent P. vivax infections that were left untreated progressed to clinical disease over 6 weeks of follow-up and became detectable by routine malaria surveillance

O Programa de Cidades Históricas (PCH) no âmbito das políticas culturais dos anos 1970: cultura, planejamento e nacional desenvolvimentismo

Este artigo analisa o Programa de Cidades Históricas (PCH) como parte da rede de relações e de poder que implementou as políticas culturais nos anos 1970 no Brasil, de modo integrado ao setor econômico do governo, com base nas linhas do projeto desenvolvimentista orientadas para o regionalismo associado à descentralização. Apresenta, inicialmente, documentos oficiais que orientaram a formulação da Política Nacional de Cultura na década de 1970 e a rede de intelectuais envolvidos na sua elaboração, contemplando diferentes agências do Estado, historicamente atuantes no campo da cultura. Destaca em tais documentos a naturalização da associação entre turismo, cultura e patrimônio, via pela qual se deu a incorporação do campo da cultura nas discussões sobre o desenvolvimento socioeconômico brasileiro. Analisa as parcerias estabelecidas entre o MEC e a Secretaria de Planejamento da Presidência da República para a criação e implantação do PCH, compreendido como um projeto de ações interministeriais integradas. Busca evidenciar os modos de incorporação das políticas culturais nos planos de desenvolvimento do Brasil Grande e apontamos a complexificação das redes e das representações acerca dos valores da brasilidade associados ao tradicional popular nordestino. Foram trazidas para a análise memórias de protagonistas do alto escalão do regime militar e de técnicos que compartilharam vivências, relacionadas àquela conjuntura e ao próprio PCH, cotejadas com uma diversificada documentação.The article analyzes the Program of Historical Cities - PCH (Programa de Cidades Históricas) as part of the network of relations and power that implemented cultural policies in the 1970s in Brazil, in an integrated manner to the government's economic sector, based on the lines of development project, oriented to a regionalism associated with decentralization. It presents first official documents that guided the formulation of the National Cultural Policy in the 1970s and the intellectual network involved in its preparation, comprising officers from different state agencies, historically active in the cultural field. The article highlights the naturalization of the association between tourism, culture and heritage and the inclusion of the cultural field in the discussions about the Brazilian socioeconomic development. It analyzes partnerships between the Ministry of Education and Culture and the Secretariat of Planning of the Presidence of the Republic (Secretaria de Planejamento da Presidência da República) for the creation and implementation of the PCH, understood as a project of integrated ministerial actions. We identify the ways of incorporation of cultural policies in development plans of the ideological program called "Great Brasil" (Brasil Grande), and pointed out the complexity of networks and representations about the Brazilianness values associated with the traditional folk Northeastern. We brought for analysis the memories of high-ranking military regime protagonists and technicians who shared experiences related to the PCH, and collated them with a diverse documentation

Agrotóxicos e saúde: realidade e desafios para mudança de práticas na agricultura.

Este artigo apresenta os resultados de uma pesquisa que teve como objetivo principal caracterizar o contexto e as práticas relacionadas ao uso de agrotóxicos entre agricultores residentes na Serrinha do Mendanha, comunidade agrícola situada em Campo Grande, município do Rio de Janeiro. A proposta metodológica teve uma abordagem qualitativa, sendo a principal fonte de dados a aplicação de questionário semiestruturado. Participaram da pesquisa 38 agricultores familiares. O perfil socioeconômico mostra uma realidade de idosos no trabalho agrícola, percentuais expressivos de analfabetismo e baixa renda. O uso de agrotóxicos é uma rotina para essas famílias. Apesar de existir uma naturalização do uso, a maioria dos informantes acredita que agrotóxicos podem afetar sua saúde, apresenta preocupação com o consumidor e gostaria de conhecer formas alternativas de cultivo. Tais achados apontam para a possibilidade de busca de mudança de práticas. Não se trata de repasse de conhecimento, pois este tipo de informação de alguma forma eles já possuem. Trata–se de desenvolver estratégias objetivando a minimização dos riscos ou mesmo o desuso de agrotóxicos na lavoura

First report of multiple lineages of dengue viruses type 1 in Rio de Janeiro, Brazil

<p>Abstract</p> <p>Background</p> <p>In Brazil dengue has been a major public health problem since DENV-1 introduction and spread in 1986. After a low or silent co-circulation, DENV-1 re-emerged in 2009 causing a major epidemic in the country in 2010 and 2011. In this study, the phylogeny of DENV-1 strains isolated in RJ after its first introduction in 1986 and after its emergence in 2009 and 2010 was performed in order to document possible evolutionary patterns or introductions in a re-emergent virus.</p> <p>Findings</p> <p>The analysis of the E gene sequences demonstrated that DENV-1 isolated during 2009/2010 still belong to genotype V (Americas/Africa) but grouping in a distinct clade (lineage II) of that represented by earlier DENV-1 (lineage I). However, strains isolated in 2011 grouped together forming another distinct clade (lineage III).</p> <p>Conclusions</p> <p>The monitoring of DENV is important to observe the spread of potentially virulent strains as well to evaluate its impact over the population during an outbreak. Whether explosive epidemics reported in Brazil caused mainly by DENV-1 was due to lineage replacement, or due the population susceptibility to this serotype which has not circulated for almost a decade or even due to the occurrence of secondary infections in a hyperendemic country, is not clear. This is the first report of multiple lineages of DENV-1 detected in Brazil.</p

Genetic diversity and structure of Iberian Peninsula cowpeas compared to world-wide cowpea accessions using high density SNP markers

Cowpea (Vigna unguiculata L. Walp) is an important legume crop due to its high protein content, adaptation to heat and drought and capacity to fix nitrogen. Europe has a deficit of cowpea production. Knowledge of genetic diversity among cowpea landraces is important for the preservation of local varieties and is the basis to obtain improved varieties. The aims of this study were to explore diversity and the genetic structure of a set of Iberian Peninsula cowpea accessions in comparison to a worldwide collection and to infer possible dispersion routes of cultivated cowpea.This study was supported by EUROLEGUME project. This project has received funding from the European Union’s Seventh Framework Programme for research, technological development and demonstration under grant agreement no 613781. European Investment Funds by FEDER/COMPETE/ POCI – Operational Competitiveness and Internationalization Programme, under Project POCI-01-0145-FEDER-006958 and National Funds by FCT – Portuguese Foundation for Science and Technology, under the project UID/AGR/04033/2013. MMA was partially supported by the Feed the Future Innovation Lab for Climate Resilient Cowpea (USAID Cooperative Agreement AID-OAA-A-13-00070), which is directed by TJC. The funding entities had no role in the design of the study, collection, analysis and interpretation of data, or in writing the manuscript.info:eu-repo/semantics/publishedVersio