ABSTRACT
The 3-chymotrypsin-like cysteine protease (3CLpro) of severe acute respiratory syndrome conoravirus 2 (SARS-CoV-2) remains a promising therapeutic target to combat COVID-19. Our group recently described a novel duplexed biochemical assay that combines self-assembled monolayers of alkanethiolates on gold with matrix assisted laser desorption ionization (MALDI) time of flight (TOF) mass spectrometry (MS) to simultaneously measure 3CLpro and human rhinovirus 3C protease activities. This study describes applying the assay for the completion of a high-throughput duplexed screen of 300,000 diverse, drug-like small molecules in 3 days. The hits were confirmed and evaluated in dose response analyses against recombinant 3CLpro, HRV3C, and the human Cathepsin L proteases. The 3CLpro specific inhibitors were further assessed for activity in cellular cytotoxicity and anti-viral assays. Structure activity relationship studies informed on structural features required for activity and selectivity to 3CLpro over HRV3C. These results will guide the optimization of 3CLpro selective inhibitors to combat COVID-19 along with antiviral compounds against coronaviruses and rhinoviruses.
Subject(s)
COVID-19 Drug Treatment , SARS-CoV-2 , Antiviral Agents/chemistry , Antiviral Agents/pharmacology , Humans , Mass Spectrometry , Protease Inhibitors/chemistry , Protease Inhibitors/pharmacology , RhinovirusABSTRACT
Hypercapnia (HC), elevation of the partial pressure of CO2 in blood and tissues, is a risk factor for mortality in patients with severe acute and chronic lung diseases. We previously showed that HC inhibits multiple macrophage and neutrophil antimicrobial functions and increases the mortality of bacterial pneumonia in mice. In this study, we show that normoxic HC increases viral replication, lung injury, and mortality in mice infected with influenza A virus (IAV). Elevated CO2 increased IAV replication and inhibited antiviral gene and protein expression in macrophages in vivo and in vitro. HC potentiated IAV-induced activation of Akt, whereas specific pharmacologic inhibition or short hairpin RNA knockdown of Akt1 in alveolar macrophages blocked HC's effects on IAV growth and the macrophage antiviral response. Our findings suggest that targeting Akt1 or the downstream pathways through which elevated CO2 signals could enhance macrophage antiviral host defense and improve clinical outcomes in hypercapnic patients with advanced lung disease.
Subject(s)
Hypercapnia/immunology , Influenza A virus/physiology , Influenza, Human/immunology , Lung/pathology , Macrophages/immunology , Oncogene Protein v-akt/metabolism , Orthomyxoviridae Infections/immunology , Animals , Cells, Cultured , Gene Expression Regulation , Humans , Immunity, Cellular , Immunosuppression Therapy , Lung/virology , Macrophage Activation , Mice , Mice, Inbred C57BL , Signal Transduction , Virus ReplicationABSTRACT
Tendo em vista as transformações nas macroestruturas e seus impactos sobre a forma como os indivíduos - jovens ou não - relacionam-se com o trabalho e o futuro (especialmente o profissional) nas sociedades capitalistas, este artigo analisa a maneira como os jovens universitários de estratos médios do Rio de Janeiro atuam e se posicionam diante da demanda de se tornarem produtivos, bem como o lugar e as condições de construção de um projeto profissional no contemporâneo. Utilizou-se o paradigma metodológico de pesquisa-ação, cuja abordagem empírica constituiu-se de uma intervenção junto a jovens estudantes de duas universidades, no âmbito de uma disciplina especificamente criada para se discutir o projeto de vida profissional. Entre os principais resultados desse estudo, destacam-se as mudanças relativas à construção de um projeto profissional baseado em princípios de racionalização, antecipação e adequação. O projeto passa a ter algum sentido se entendido como forma de construção, interlocução e representação de desejos de realização dos indivíduos, num processo que os leva a reexaminarem suas possibilidades e limitações, e sua própria posição diante da vida. Neste reexame as incertezas dos jovens diante das novas engrenagens produtivas são apreendidas como fonte de compreensão da inter-relação que estabelecem com a conjuntura social, e não apenas como algo que deva ser corrigido ou eliminado(AU)
In the context of long-standing macro-structural transformations and of their impact on how individuals - young or not - relate themselves to work and to their professional future in capitalist societies, this article analyses how young undergraduate students of the middle classes in Rio de Janeiro, respond to the societal demand of becoming professionally active and productive. It also focuses on the conditions which allow for the construction of a professional future. A methodological paradigm of action-research was used, whose empirical approach involved an intervention project in which these young people studying at two different universities were invited to discuss their professional project in a discipline oriented towards this aim. Among the results of the project could be found changes concerning the construction of a professional future based on principles such as, rationalization, anticipation and modeling. It was observed that the professional project begins to make sense once it is understood as a form of construction, interlocution and representation of the objectives and desires of the individuals, a process that leads them to revisit and examine their possibilities and limitations, as well as their subjective position in life. Alongside this examination, young people's uncertainties concerning macro productive mechanisms and outcomes can be understood as a source of help to position them in the overall productive conjuncture, rather than something to be corrected or eliminated(AU)
Subject(s)
Young Adult , Project Formulation , Professional Role/psychology , Work/psychologyABSTRACT
Tendo em vista as transformações nas macroestruturas e seus impactos sobre a forma como os indivíduos - jovens ou não - relacionam-se com o trabalho e o futuro (especialmente o profissional) nas sociedades capitalistas, este artigo analisa a maneira como os jovens universitários de estratos médios do Rio de Janeiro atuam e se posicionam diante da demanda de se tornarem produtivos, bem como o lugar e as condições de construção de um projeto profissional no contemporâneo. Utilizou-se o paradigma metodológico de pesquisa-ação, cuja abordagem empírica constituiu-se de uma intervenção junto a jovens estudantes de duas universidades, no âmbito de uma disciplina especificamente criada para se discutir o projeto de vida profissional. Entre os principais resultados desse estudo, destacam-se as mudanças relativas à construção de um projeto profissional baseado em princípios de racionalização, antecipação e adequação. O projeto passa a ter algum sentido se entendido como forma de construção, interlocução e representação de desejos de realização dos indivíduos, num processo que os leva a reexaminarem suas possibilidades e limitações, e sua própria posição diante da vida. Neste reexame as incertezas dos jovens diante das novas engrenagens produtivas são apreendidas como fonte de compreensão da inter-relação que estabelecem com a conjuntura social, e não apenas como algo que deva ser corrigido ou eliminado.
In the context of long-standing macro-structural transformations and of their impact on how individuals - young or not - relate themselves to work and to their professional future in capitalist societies, this article analyses how young undergraduate students of the middle classes in Rio de Janeiro, respond to the societal demand of becoming professionally active and productive. It also focuses on the conditions which allow for the construction of a professional future. A methodological paradigm of action-research was used, whose empirical approach involved an intervention project in which these young people studying at two different universities were invited to discuss their professional project in a discipline oriented towards this aim. Among the results of the project could be found changes concerning the construction of a professional future based on principles such as, rationalization, anticipation and modeling. It was observed that the professional project begins to make sense once it is understood as a form of construction, interlocution and representation of the objectives and desires of the individuals, a process that leads them to revisit and examine their possibilities and limitations, as well as their subjective position in life. Alongside this examination, young people's uncertainties concerning macro productive mechanisms and outcomes can be understood as a source of help to position them in the overall productive conjuncture, rather than something to be corrected or eliminated.
