Silibinin and SARS-CoV-2: Dual Targeting of Host Cytokine Storm and Virus Replication Machinery for Clinical Management of COVID-19 Patients

COVID-19, the illness caused by infection with the novel coronavirus SARS-CoV-2, is a rapidly spreading global pandemic in urgent need of effective treatments. Here we present a comprehensive examination of the host- and virus-targeted functions of the flavonolignan silibinin, a potential drug candidate against COVID-19/SARS-CoV-2. As a direct inhibitor of STAT3-a master checkpoint regulator of inflammatory cytokine signaling and immune response-silibinin might be expected to phenotypically integrate the mechanisms of action of IL-6-targeted monoclonal antibodies and pan-JAK1/2 inhibitors to limit the cytokine storm and T-cell lymphopenia in the clinical setting of severe COVID-19. As a computationally predicted, remdesivir-like inhibitor of RNA-dependent RNA polymerase (RdRp)-the central component of the replication/transcription machinery of SARS-CoV-2-silibinin is expected to reduce viral load and impede delayed interferon responses. The dual ability of silibinin to target both the host cytokine storm and the virus replication machinery provides a strong rationale for the clinical testing of silibinin against the COVID-19 global public health emergency. A randomized, open-label, phase II multicentric clinical trial (SIL-COVID19) will evaluate the therapeutic efficacy of silibinin in the prevention of acute respiratory distress syndrome in moderate-to-severe COVID-19-positive onco-hematological patients at the Catalan Institute of Oncology in Catalonia, Spain

Comparación de dos morfotipos de Zuccagnia punctata (Fabaceae) en los Valles Calchaquíes en Argentina

Zuccagnia punctata Cav. (Fabaceae) is a medicinal aromatic shrub, a monotypic species with a wide distribution in Argentina. Two morphotypes were found in Valles Calchaquíes, one with yellow fruits (YF) and other with red-brown fruits (RBF). The color of fruits varies between individuals within a population, with some plants producing red-brown fruits and other yellow fruits. The cytogenetic differentiation was determined using root tips. Leaves and fruits were used to analyze chemical composition by HPLC-DAD and HPLC/MS/MS. The number of chromosome (2n = 24) was reported for the first time for the genus. The yellow fruits and red-brown fruits morphotypes showed different karyotypes. The compounds common to both morphotypes were identified in leaves and fruits as chalcones, compounds with biological activity and biomarkers used for quality control of Z. punctata products. Two chemotypes related to the color of the fruits, were detected. An anthocyanin, cyanidin 3-glucoside was found in red-brown fruits (4.75 mg.g-1 fruits) but not in yellow fruits. These is the first report on yellow fruits morphotype of Z. punctata and on presence of anthocyanins in Z. punctata red-brown fruits morphotype. This work contributes to the knowledge of Z. punctata, a native plant from Argentina with high economic potential to promote the regional economy.Zuccagnia punctata Cav. (Fabaceae) es un arbusto aromático medicinal monotípico con amplia distribución en Argentina. En los Valles Calchaquíes se encontraron dos morfotipos, uno con frutos amarillos y otro con frutos marrón rojizo. El color de los frutos varía entre individuos de una misma población, observándose plantas que tienen frutos de color marrón rojizo y otras con frutos de color amarillo. La diferenciación citogenética fue determinada usando puntas de raíces mientras que las hojas y frutos se usaron para analizar la composición química por HPLC-DAD y HPLC/MS/MS. Se informó por primera vez el número cromosómico (2n = 24) para el género Zuccagnia. Ambos morfotipos mostraron diferentes cariotipos. Se identificaron chalconas como compuestos comunes en las hojas y frutos de ambos morfotipos. Las chalconas son consideradas biomarcadores en el control de calidad de productos a base de Z. punctata. Se detectaron dos quimiotipos relacionados al color de los frutos. La antocianina, cianidin 3-glucosido se encontró en frutos marrón rojizos (4.75 mg.g-1 fruto) pero no en frutos amarillos. Este es el primer informe sobre un morfotipo de Z. punctata con frutos amarillos y sobre la presencia de antocianinas en el morfotipo de frutos rojos. Este trabajo contribuye al conocimiento de Z. punctata, una planta nativa de Argentina con alto potencial económico para promover el desarrollo regional.Fil: Álvarez, María A.. Universidad Nacional de Tucumán. Instituto de Bioprospección y Fisiología Vegetal. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto de Bioprospección y Fisiología Vegetal; ArgentinaFil: Correa Uriburu, Florencia Maria. Universidad Nacional de Tucumán. Instituto de Bioprospección y Fisiología Vegetal. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto de Bioprospección y Fisiología Vegetal; ArgentinaFil: Barrera, María Celina. Universidad Nacional de Tucumán. Instituto de Bioprospección y Fisiología Vegetal. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto de Bioprospección y Fisiología Vegetal; ArgentinaFil: Enrico, Roxana Judith. Universidad Nacional de Tucumán. Instituto de Bioprospección y Fisiología Vegetal. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto de Bioprospección y Fisiología Vegetal; Argentina. Universidad Nacional de Tucumán. Facultad de Ciencias Naturales e Instituto Miguel Lillo; ArgentinaFil: Andrada, Aldo Rubén. Instituto de Genetica y Microbiologia ; Direccion de Biologia Integrativa ; Fundacion Miguel Lillo;Fil: Silenzi Usandivaras, Gabriela M.. Universidad Nacional de Tucumán. Facultad de Ciencias Naturales e Instituto Miguel Lillo; Argentina. Instituto de Genetica y Microbiologia ; Direccion de Biologia Integrativa ; Fundacion Miguel Lillo;Fil: Páez, Valeria A.. Instituto de Genetica y Microbiologia ; Direccion de Biologia Integrativa ; Fundacion Miguel Lillo;Fil: Caro, María S.. Universidad Nacional de Tucumán. Facultad de Ciencias Naturales e Instituto Miguel Lillo; Argentina. Instituto de Genetica y Microbiologia ; Direccion de Biologia Integrativa ; Fundacion Miguel Lillo;Fil: Zampini, Iris Catiana. Universidad Nacional de Tucumán. Instituto de Bioprospección y Fisiología Vegetal. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto de Bioprospección y Fisiología Vegetal; Argentina. Universidad Nacional de Tucumán. Facultad de Ciencias Naturales e Instituto Miguel Lillo; ArgentinaFil: Isla, Maria Ines. Universidad Nacional de Tucumán. Instituto de Bioprospección y Fisiología Vegetal. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Instituto de Bioprospección y Fisiología Vegetal; Argentina. Universidad Nacional de Tucumán. Facultad de Ciencias Naturales e Instituto Miguel Lillo; Argentin

Low-Scale Expression and Purification of an Active Putative Iduronate 2-Sulfate Sulfatase-Like Enzyme from Escherichia coli K12

The sulfatase family involves a group of enzymes with a large degree of similarity. Until now, sixteen human sulfatases have been identified, most of them found in lysosomes. Human deficiency of sulfatases generates various genetic disorders characterized by abnormal accumulation of sulfated intermediate compounds. Mucopolysaccharidosis type II is characterized by the deficiency of iduronate 2-sulfate sulfatase (IDS), causing the lysosomal accumulation of heparan and dermatan sulfates. Currently, there are several cases of genetic diseases treated with enzyme replacement therapy, which have generated a great interest in the development of systems for recombinant protein expression. In this work we expressed the human recombinant IDS-Like enzyme (hrIDS-Like) in Escherichia coli DH5α. The enzyme concentration revealed by ELISA varied from 78. 13 to 94. 35 ng/ml and the specific activity varied from 34. 20 to 25. 97 nmol/h/mg. Western blotting done after affinity chromatography purification showed a single band of approximately 40 kDa, which was recognized by an IgY polyclonal antibody that was developed against the specific peptide of the native protein. Our 100 ml-shake-flask assays allowed us to improve the enzyme activity seven fold, compared to the E. coli JM109/pUC13-hrIDS-Like system. Additionally, the results obtained in the present study were equal to those obtained with the Pichia pastoris GS1115/pPIC-9-hrIDS-Like system (3 L bioreactor scale). The system used in this work (E. coli DH5α/pGEX-3X-hrIDS-Like) emerges as a strategy for improving protein expression and purification, aimed at recombinant protein chemical characterization, future laboratory assays for enzyme replacement therapy, and as new evidence of active putative sulfatase production in E. coli. © 2013 The Microbiological Society of Korea and Springer-Verlag Berlin Heidelberg.Fil: Morales Álvarez, Edwin David. Universidad del Quindio; ColombiaFil: Rivera Hoyos, Claudia Marcela. Universidad del Quindio. Facultad de Medicina; ColombiaFil: Baena Moncada, Angelica Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Landázuri, Patricia. Universidad del Quindio. Facultad de Medicina. Centro de Investig. Biomédicas; ColombiaFil: Poutou Piñales, Raúl A.. Pontificia Universidad Javeriana; ColombiaFil: Sáenz Suárez, Homero. Universidad del Quindio; ColombiaFil: Barrera, Luis A.. Pontificia Universidad Javeriana; ColombiaFil: Echeverri Peña, Olga Y.. Pontificia Universidad Javeriana; Colombi

Instantaneous transport of salt, nutrients, suspended matter and chlorophyll-a in the tropical estuarine system of Santos

A contribuição dos canais estuarinos de Santos e São Vicente para a eutrofização da baía de Santos foi avaliada quantificando-se o transporte instantâneo de sal, fosfato e nitrogênio inorgânico dissolvido (NID), material em suspensão orgânico (MSO) e inorgânico (MSI) e clorofila-a, durante a estação seca (inverno austral- Agosto/1999) e chuvosa (verão austral- Janeiro/ 2000). As amostragens foram realizadas em períodos de sizígia e quadratura, durante as marés enchentes e vazantes, nas secções transversais das bocas dos canais de São Vicente e Santos. Os valores de transporte instantâneo obtidos durante o período de amostragem indicaram exportação de NID, principalmente sobre a forma de N-NH4 (valor máximo de 1155,1 g s-1) na estação chuvosa; importação de fosfato durante a estação seca (máximo de 385,6 g s-1) e exportação de MSI, MSO e clorofila-a em períodos de maior contribuição fluvial. Estes resultados indicam uma importante contribuição dos canais estuarinos de Santos e São Vicente para a eutrofização da baía de Santos, especialmente durante o período chuvoso.The contribution of the polluted São Vicente and Santos estuarine channels to the eutrophication of Santos bay was assessed through the quantification of instantaneous transport of salt, dissolved inorganic nitrogen (DIN) and phosphate, organic and inorganic matter (OSM and ISM) and chlorophyll-a (Chl-a), during dry (austral winter- August/ 1999) and rainy (austral summer- January/2000) seasons. Samplings were carried out during spring and neap tides, in flood and ebb phases, in two transversal sections at the mouths of the São Vicente and Santos channels. Instantaneous transport values generally indicated importation of salt to the estuarine channels, exportation of DIN to the bay, mainly as N-NH4, at a maximum rate of 1155.1 g s-1 during the rainy season; importation of phosphate during the dry season (maximum of 385 g s-1) and exportation of ISM, OSM and Chl-a during periods of greater freshwater discharge. These results demonstrate the great contribution made by the Santos and São Vicente estuaries to the eutrophication of Santos bay, especially in the rainy season

Silibinin Suppresses Tumor Cell-Intrinsic Resistance to Nintedanib and Enhances Its Clinical Activity in Lung Cancer

The anti-angiogenic agent nintedanib has been shown to prolong overall and progression-free survival in patients with advanced non-small-cell lung cancer (NSCLC) who progress after first-line platinum-based chemotherapy and second-line immunotherapy. Here, we explored the molecular basis and the clinical benefit of incorporating the STAT3 inhibitor silibinin-a flavonolignan extracted from milk thistle-into nintedanib-based schedules in advanced NSCLC. First, we assessed the nature of the tumoricidal interaction between nintedanib and silibinin and the underlying relevance of STAT3 activation in a panel of human NSCLC cell lines. NSCLC cells with poorer cytotoxic responses to nintedanib exhibited a persistent, nintedanib-unresponsive activated STAT3 state, and deactivation by co-treatment with silibinin promoted synergistic cytotoxicity. Second, we tested whether silibinin could impact the lysosomal sequestration of nintedanib, a lung cancer cell-intrinsic mechanism of nintedanib resistance. Silibinin partially, but significantly, reduced the massive lysosomal entrapment of nintedanib occurring in nintedanib-refractory NSCLC cells, augmenting the ability of nintedanib to reach its intracellular targets. Third, we conducted a retrospective, observational multicenter study to determine the efficacy of incorporating an oral nutraceutical product containing silibinin in patients with NSCLC receiving a nintedanib/docetaxel combination in second- and further-line settings (n = 59). Overall response rate, defined as the combined rates of complete and partial responses, was significantly higher in the study cohort receiving silibinin supplementation (55%) than in the control cohort (22%, p = 0.011). Silibinin therapy was associated with a significantly longer time to treatment failure in multivariate analysis (hazard ratio 0.43, p = 0.013), despite the lack of overall survival benefit (hazard ratio 0.63, p = 0.190). Molecular mechanisms dictating the cancer cell-intrinsic responsiveness to nintedanib, such as STAT3 activation and lysosomal trapping, are amenable to pharmacological intervention with silibinin. A prospective, powered clinical trial is warranted to confirm the clinical relevance of these findings in patients with advanced NSCLC

Identification of a cytokine network sustaining neutrophil and Th17 activation in untreated early rheumatoid arthritis

© 2010 Cascão et al.; licensee BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.Introduction: Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune disease characterized by sustained synovitis. Recently, several studies have proposed neutrophils and Th17 cells as key players in the onset and perpetuation of this disease. The main goal of this work was to determine whether cytokines driving neutrophil and Th17 activation are dysregulated in very early rheumatoid arthritis patients with less than 6 weeks of disease duration and before treatment (VERA). Methods: Cytokines related to neutrophil and Th17 activation were quantified in the serum of VERA and established RA patients and compared with other very early arthritis (VEA) and healthy controls. Synovial fluid (SF) from RA and osteoarthritis (OA) patients was also analyzed. Results: VERA patients had increased serum levels of cytokines promoting Th17 polarization (IL-1b and IL-6), as well as IL-8 and Th17-derived cytokines (IL-17A and IL-22) known to induce neutrophil-mediated inflammation. In established RA this pattern is more evident within the SF. Early treatment with methotrexate or corticosteroids led to clinical improvement but without an impact on the cytokine pattern. Conclusions: VERA patients already display increased levels of cytokines related with Th17 polarization and neutrophil recruitment and activation, a dysregulation also found in SF of established RA. 0 Thus, our data suggest that a cytokine-milieu favoring Th17 and neutrophil activity is an early event in RA pathogenesis.This work was supported by a grant from Sociedade Portuguesa de Reumatologia/Schering-Plough 2005. RAM and RC were funded by Fundação para a Ciência e a Tecnologia (FCT) SFRH/BD/30247/2006 and SFRH/BD/40513/2007, respectively. MMS-C was funded by Marie Curie Intra-European Fellowship PERG-2008-239422 and a EULAR Young Investigator Award

Transactive Response DNA-Binding Protein (TARDBP/TDP-43) Regulates Cell Permissivity to HIV-1 Infection by Acting on HDAC6

The transactive response DNA-binding protein (TARDBP/TDP-43) influences the processing of diverse transcripts, including that of histone deacetylase 6 (HDAC6). Here, we assessed TDP-43 activity in terms of regulating CD4+ T-cell permissivity to HIV-1 infection. We observed that overexpression of wt-TDP-43 increased both mRNA and protein levels of HDAC6, resulting in impaired HIV-1 infection independently of the viral envelope glycoprotein complex (Env) tropism. Consistently, using an HIV-1 Env-mediated cell-to-cell fusion model, the overexpression of TDP-43 levels negatively affected viral Env fusion capacity. Silencing of endogenous TDP-43 significantly decreased HDAC6 levels and increased the fusogenic and infection activities of the HIV-1 Env. Using pseudovirus bearing primary viral Envs from HIV-1 individuals, overexpression of wt-TDP-43 strongly reduced the infection activity of Envs from viremic non-progressors (VNP) and rapid progressors (RP) patients down to the levels of the inefficient HIV-1 Envs observed in long-term non-progressor elite controllers (LTNP-EC). On the contrary, silencing endogenous TDP-43 significantly favored the infectivity of primary Envs from VNP and RP individuals, and notably increased the infection of those from LTNP-EC. Taken together, our results indicate that TDP-43 shapes cell permissivity to HIV-1 infection, affecting viral Env fusion and infection capacities by altering the HDAC6 levels and associated tubulin-deacetylase anti-HIV-1 activity.This work is supported by the Spanish AIDS network “Red Temática Cooperativa de Investigación en SIDA” RD12/0017/0002, RD12/0017/0028, RD12/0017/0034, RD16/0025/0011, RDCIII16/0002/0005 and RD16/0025/0041 as part of the Plan Nacional R + D+I and co-funded by the Spanish “Instituto de Salud Carlos III (ISCIII)-Subdirección General de Evaluación y el Fondo Europeo de Desarrollo Regional (FEDER)”. J.B. is a researcher from “Fundació Institut de Recerca en Ciències de la Salut Germans Trias i Pujol” supported by the Health Department of the Catalonian Government/Generalitat de Catalunya and ISCIII grant numbers PI17/01318 and PI20/00093 (to J.B.). Work in CC Lab was supported by grants SAF (2010-17226) and (2016-77894-R) from MINECO (Spain), FIS (PI 13/02269, ISCIII) and PI20/00093. Work in CF Lab was supported by the Cabildo Insular de Tenerife (grants CGIEU0000219140 and “Apuestas científicas del ITER para colaborar en la lucha contra la COVID-19”); the agreement with the Instituto Tecnológico y de Energías Renovables (ITER) to strengthen scientific and technological education, training research, development and innovation in Genomics, Personalized Medicine and Biotechnology (grant number OA17/008). A.V.-F.’s Lab is supported by the European Regional Development Fund (ERDF), RTI2018-093747-B-100 (“Ministerio de Ciencia e Innovación”, Spain), “Ministerio de Ciencia, Innovación y Universidades” (Spain), ProID2020010093 (“Agencia Canaria de Investigación, Innovación y Sociedad de la Información” and European Social Fund), UNLL10-3E-783 (ERDF and “Fundación CajaCanarias”) and “SEGAI-ULL”. S.P-Y is funded by “Fundación Doctor Manuel Morales” (La Palma, Spain) and “Contrato Predoctoral Ministerio-ULL Formación de Doctores” (2019 Program) (“Ministerio de Ciencia, Innovación y Universidades”, Spain). R.C.-R. is funded by RD16/0025/0011 and ProID2020010093 (“Agencia Canaria de Investigación, Innovación y Sociedad de la Información” and European Social Fund). J.G.-L. is funded by the “Juan de la Cierva de Incorporación” Spanish Program (IJC2019-038902-I) (“Ayudas Juan de la Cierva de incorporación; Agencia Estatal de Investigación. Ministerio de Ciencia e Innovación”).S