Targeting proteases involved in the viral replication of SARS-CoV-2 by sesquiterpene lactones from chicory (Cichorium intybus L.)

Funding Information: This research has been supported by the EU Horizon 2020 research & innovation programme (H2020-NMBP-BIO-2017, project CHIC grant agreement no. 760891). Publisher Copyright: © 2022 The Royal Society of Chemistry.SARS-CoV-2 is a highly transmissible and pathogenic coronavirus causing a respiratory disease that emerged in 2019, leading to a public health emergency situation which continues to date. The treatment options are still very limited and vaccines available are less effective against new variants. SARS-CoV-2 enzymes, namely main protease (Mpro) and papain-like protease (PLpro), play a pivotal role in the viral life cycle, making them a putative drug target. Here, we described for the first time the potential inhibitory activity of chicory extract against both proteases. Besides, we have identified that the four most abundant sesquiterpene lactones in chicory inhibited these proteases, showing an effective binding in the active sites of Mpro and PLpro. This paper provides new insight for further drug development or food-based strategies for the prevention of SARS-CoV-2 by targeting viral proteases.publishersversionepub_ahead_of_prin

Supercritical CO2 Extraction as a Tool to Isolate Anti-Inflammatory Sesquiterpene Lactones from Cichorium intybus L. Roots

Funding: This work was conducted under the “CHIC” project (H2020-NMBP-BIO-2017) with financial support received from the EU Horizon 2020 research and innovation programme under grant agreement N. 760891.Cichorium intybus L. or chicory plants are a natural source of health-promoting compounds in the form of supplements such as inulin, as well as other bioactive compounds such as sesquiterpene lactones (SLs). After inulin extraction, chicory roots are considered waste, with most SLs not being harnessed. We developed and optimized a new strategy for SL extraction that can contribute to the conversion of chicory root waste into valuable products to be used in human health-promoting applications. In our work, rich fractions of SLs were recovered from chicory roots using supercritical CO2. A response surface methodology was used to optimize the process parameters (pressure, temperature, flow rate, and co-solvent percentage) for the extraction performance. The best operating conditions were achieved at 350 bar, 40 °C, and 10% EtOH as a co-solvent in a 15 g/min flow rate for 120 min. The extraction with supercritical CO2 revealed to be more selective for the SLs than the conventional solid-liquid extraction with ethyl acetate. In our work, 1.68% mass and a 0.09% sesquiterpenes yield extraction were obtained, including the recovery of two sesquiterpene lactones (8-deoxylactucin and 11β,13-dihydro-8-deoxylactucin), which, to the best of our knowledge, are not commercially available. A mixture of the abovementioned compounds were tested at different concentrations for their toxic profile and anti-inflammatory potential towards a human calcineurin/NFAT orthologue pathway in a yeast model, the calcineurin/Crz1 pathway. The SFE extract obtained, rich in SLs, yielded results of inhibition of 61.74 ± 6.87% with 50 µg/mL, and the purified fraction containing 8-deoxylactucin and 11β,13-dihydro-8-deoxylactucin inhibited the activation of the reporter gene up to 53.38 ± 3.9% at 10 µg/mL. The potential activity of the purified fraction was also validated by the ability to inhibit Crz1 nuclear translocation and accumulation. These results reveal a possible exploitable green technology to recover potential anti-inflammatory compounds from chicory roots waste after inulin extraction.publishersversionpublishe

Assessing the intestinal permeability and anti-inflammatory potential of sesquiterpene lactones from chicory

Funding: This research and the article processing cost were funded by EU Horizon 2020 research & innovation programme under grant agreement N. 760891 project CHIC. M.S.M. also acknowledges the financial support from Fundação para a Ciência e Tecnologia for her PhD scholarship (SFRD/BD/145551/2019).Cichorium intybus L. has recently gained major attention due to large quantities of health-promoting compounds in its roots, such as inulin and sesquiterpene lactones (SLs). Chicory is the main dietary source of SLs, which have underexplored bioactive potential. In this study, we assessed the capacity of SLs to permeate the intestinal barrier to become physiologically available, using in silico predictions and in vitro studies with the well-established cell model of the human intestinal mucosa (differentiated Caco-2 cells). The potential of SLs to modulate inflammatory responses through modulation of the nuclear factor of activated T-cells (NFAT) pathway was also evaluated, using a yeast reporter system. Lactucopicrin was revealed as the most permeable chicory SL in the intestinal barrier model, but it had low anti-inflammatory potential. The SL with the highest anti-inflammatory potential was 11β,13-dihydrolactucin, which inhibited up to 54% of Calcineurin-responsive zinc finger (Crz1) activation, concomitantly with the impairment of the nuclear accumulation of Crz1, the yeast orthologue of human NFAT.publishersversionpublishe

Postmortem Findings for 7 Neonates with Congenital Zika Virus Infection

Postmortem examination of 7 neonates with congenital Zika virus infection in Brazil revealed microcephaly, ventriculomegaly, dystrophic calcifications, and severe cortical neuronal depletion in all and arthrogryposis in 6. Other findings were leptomeningeal and brain parenchymal inflammation and pulmonary hypoplasia and lymphocytic infiltration in liver and lungs. Findings confirmed virus neurotropism and multiple organ infection

Expressão nuclear do P53 em carcinoma de células transicionais da bexiga Nuclear expression of P53 protein in transitional cell carcinoma of the bladder

OBJETIVO: Investigar a expressão imuno-histoquímica da p53 com fator de risco em carcinoma de células transicionais da bexiga (CCT). MÉTODOS: Foram estudados restrospectivamente 90 pacientes com CCT com idade média de 71 anos: G1 - 45, G2 - 29, G3 - 16, pTa-1 - 62 e pT2-4 - 28. Entre os pacientes com tumores não invasivos houve recidiva vesical em 35 casos (55,5%). Os tumores superficiais foram tratados por ressecção trans-uretral associados ao BCG (>G1), e os invasivos por cistectomia radical. O tempo médio de seguimento dos pacientes foi de 55 meses e 25 deles faleceram da doença. A expressão imuno-histoquímica foi estudada em peças preservadas em formol 10% e blocos de parafina pelo método da avidina-biotina-imunoperoxidase. Considerou-se p53 positivo o tumor com índice de marcação nuclear superior a 10%. RESULTADOS: A expressão da p53 mostrou associação com o grau do tumor e com o estádio da lesão primária (p=0,01), mas não com o tamanho do tumor vesical (p=0,25) ou com a taxa de recidiva dos tumores superficiais (p=0,81). Houve forte correlação entre o padrão de marcação da p53 com metástases (p=0,002) e com a sobrevida dos pacientes (p=0,003). CONCLUSÃO: A expressão da p53 mostrou valor preditivo para grau tumoral, estádio, incidência de metástases e sobrevida dos pacientes, mas não para recidiva vesical dos tumores superficiais.<br>OBJECTIVE: To investigate the immunoexpression of p53 protein as a risk factor in transitional cell carcinoma of the bladder (TCC). METHODS: We analyzed retrospectively 90 patients with TCC and mean age of 71 years: G1 - 45, G1 - 29, G3 - 16, pTa-1 - 62 and pT2-4 - 28. The superficial TCC were treated TUR plus intravesical BCG (>G1), and the invasives by radical cystectomy and urinary diversion. The mean time of followup was 55 months and 25 patients died of the disease. The rate or reccurence in superficial tumors was 55.5%. The p53 immunoexpression was determined in formalin fixed paraffin embedded tumors sections by the avidin-biotin-immunoperoxidase method (DO - 7, Dako). We considered p53 positive TCC when the labeling nuclear index was >10%. RESULTS: P53 immunoexpression exhibited a positive association with tumor grade and stage (p=0.01), but not with the size of primary lesion (p=0.25) or rate of reccurence of pTa-1 tumors (p=0.81). A strong correlation was seen with metastases (p=0.002) and survival (p=0.003). CONCLUSION: A positive reaction for p53 showed correlation with tumor grade, T stage, metastases rate and parient survival, but had no predictive value for local reccurence rate of superficial tumors

Cross-sectional study defines difference in malaria morbidity in two Yanomami communities on Amazonian boundary between Brazil and Venezuela

It is well established that immunity to malaria is short-lived and is maintained by the continuous contact with the parasite. We now show that the stable transmission of malaria in Yanomami Amerindian communities maintains a degree of immunity in the exposed population capable to reduce prevalence and morbidity of malaria. We examined 508 Yanomami Amerindians living along Orinoco (407) and Mucajaí (101) rivers, on the Venezuelan and Brazilian Amazon region, respectively. At Orinoco villages, malaria was hyperendemic and presented stable transmission, while at Mucajaí villages it was mesoendemic and showed unstable transmission. The frequency of Plasmodium vivax and P. falciparum was roughly comparable in Venezuelan and Brazilian communities. Malaria presented different profiles at Orinoco and Mucajaí villages. In the former communities, malaria showed a lower prevalence (16% x 40.6%), particularly among those over 10 years old (5.2% x 34.8%), a higher frequency of asymptomatic cases (38.5% x 4.9%), and a lower frequency of cases of severe malaria (9.2% x 36.5%). Orinoco villagers also showed a higher reactivity of the immune system, measured by the frequency of splenomegaly (72.4% x 29.7%) and by the splenic index (71.4% over level 1 x 28.6), and higher prevalence (91.1% x 72.1%) and mean titer (1243 x 62) of antiplasmodial IgG antibodies, as well as a higher prevalence (77.4% x 24.7%) and mean titer (120 x 35) of antiplasmodial IgM antibodies. Our findings show that in isolated Yanomami communities the stability of malaria transmission, and the consequent continuous activation of the immune system of the exposed population, leads to the reduction of malaria prevalence and morbidity