Cabbage and fermented vegetables : From death rate heterogeneity in countries to candidates for mitigation strategies of severe COVID-19

Large differences in COVID-19 death rates exist between countries and between regions of the same country. Some very low death rate countries such as Eastern Asia, Central Europe, or the Balkans have a common feature of eating large quantities of fermented foods. Although biases exist when examining ecological studies, fermented vegetables or cabbage have been associated with low death rates in European countries. SARS-CoV-2 binds to its receptor, the angiotensin-converting enzyme 2 (ACE2). As a result of SARS-CoV-2 binding, ACE2 downregulation enhances the angiotensin II receptor type 1 (AT(1)R) axis associated with oxidative stress. This leads to insulin resistance as well as lung and endothelial damage, two severe outcomes of COVID-19. The nuclear factor (erythroid-derived 2)-like 2 (Nrf2) is the most potent antioxidant in humans and can block in particular the AT(1)R axis. Cabbage contains precursors of sulforaphane, the most active natural activator of Nrf2. Fermented vegetables contain many lactobacilli, which are also potent Nrf2 activators. Three examples are: kimchi in Korea, westernized foods, and the slum paradox. It is proposed that fermented cabbage is a proof-of-concept of dietary manipulations that may enhance Nrf2-associated antioxidant effects, helpful in mitigating COVID-19 severity.Peer reviewe

Nrf2-interacting nutrients and COVID-19 : time for research to develop adaptation strategies

There are large between- and within-country variations in COVID-19 death rates. Some very low death rate settings such as Eastern Asia, Central Europe, the Balkans and Africa have a common feature of eating large quantities of fermented foods whose intake is associated with the activation of the Nrf2 (Nuclear factor (erythroid-derived 2)-like 2) anti-oxidant transcription factor. There are many Nrf2-interacting nutrients (berberine, curcumin, epigallocatechin gallate, genistein, quercetin, resveratrol, sulforaphane) that all act similarly to reduce insulin resistance, endothelial damage, lung injury and cytokine storm. They also act on the same mechanisms (mTOR: Mammalian target of rapamycin, PPAR gamma:Peroxisome proliferator-activated receptor, NF kappa B: Nuclear factor kappa B, ERK: Extracellular signal-regulated kinases and eIF2 alpha:Elongation initiation factor 2 alpha). They may as a result be important in mitigating the severity of COVID-19, acting through the endoplasmic reticulum stress or ACE-Angiotensin-II-AT(1)R axis (AT(1)R) pathway. Many Nrf2-interacting nutrients are also interacting with TRPA1 and/or TRPV1. Interestingly, geographical areas with very low COVID-19 mortality are those with the lowest prevalence of obesity (Sub-Saharan Africa and Asia). It is tempting to propose that Nrf2-interacting foods and nutrients can re-balance insulin resistance and have a significant effect on COVID-19 severity. It is therefore possible that the intake of these foods may restore an optimal natural balance for the Nrf2 pathway and may be of interest in the mitigation of COVID-19 severity

Rationale, study design, and analysis plan of the Alveolar Recruitment for ARDS Trial (ART): Study protocol for a randomized controlled trial

Background: Acute respiratory distress syndrome (ARDS) is associated with high in-hospital mortality. Alveolar recruitment followed by ventilation at optimal titrated PEEP may reduce ventilator-induced lung injury and improve oxygenation in patients with ARDS, but the effects on mortality and other clinical outcomes remain unknown. This article reports the rationale, study design, and analysis plan of the Alveolar Recruitment for ARDS Trial (ART). Methods/Design: ART is a pragmatic, multicenter, randomized (concealed), controlled trial, which aims to determine if maximum stepwise alveolar recruitment associated with PEEP titration is able to increase 28-day survival in patients with ARDS compared to conventional treatment (ARDSNet strategy). We will enroll adult patients with ARDS of less than 72 h duration. The intervention group will receive an alveolar recruitment maneuver, with stepwise increases of PEEP achieving 45 cmH(2)O and peak pressure of 60 cmH2O, followed by ventilation with optimal PEEP titrated according to the static compliance of the respiratory system. In the control group, mechanical ventilation will follow a conventional protocol (ARDSNet). In both groups, we will use controlled volume mode with low tidal volumes (4 to 6 mL/kg of predicted body weight) and targeting plateau pressure <= 30 cmH2O. The primary outcome is 28-day survival, and the secondary outcomes are: length of ICU stay; length of hospital stay; pneumothorax requiring chest tube during first 7 days; barotrauma during first 7 days; mechanical ventilation-free days from days 1 to 28; ICU, in-hospital, and 6-month survival. ART is an event-guided trial planned to last until 520 events (deaths within 28 days) are observed. These events allow detection of a hazard ratio of 0.75, with 90% power and two-tailed type I error of 5%. All analysis will follow the intention-to-treat principle. Discussion: If the ART strategy with maximum recruitment and PEEP titration improves 28-day survival, this will represent a notable advance to the care of ARDS patients. Conversely, if the ART strategy is similar or inferior to the current evidence-based strategy (ARDSNet), this should also change current practice as many institutions routinely employ recruitment maneuvers and set PEEP levels according to some titration method.Hospital do Coracao (HCor) as part of the Program 'Hospitais de Excelencia a Servico do SUS (PROADI-SUS)'Brazilian Ministry of Healt

Estudo da modulação das células mielóides supressoras no adenocarcinoma metastático após terapia antiangiogênica

Renal cell carcinoma (RCC), the third leading cause of death in genitourinary cancers. RCCs are highly vascularized and resistant to radiotherapy and chemotherapy. Antiangiogenic drugs are promising and widely used in clinical, on the other hand, mechanism of evasive resistance has been seen to treatment with anti-VEGF antibody. Recent reports suggest that treatment resistance of patients with cancer is related to the presence of myeloid-derived suppressor cells. Endostatin (ES) is a fragment of collagen XVIII that mediates antiangiogenic activity, however, its mechanisms of action are unclear. In this study, we tracked Gr-1+ cells in different organs, evaluated the role of CD11b+Gr-1+ cells and their subsets during tumoral progression and examined the therapeutic potential of ES in modulating CD11b+Gr-1+ cells and their subsets as well as their immunosuppressive activities in lung metastasis. Healthy Balb/c mice were used to detect Gr-1+ cells. CCRm-bearing mice were nephrectomized on day 7 and collected metastatic lung, spleen and bone marrow after 3, 7 and 10 days. Quantification of CD11b+Gr-1+ cells and their monocytic and granulocytic subsets was performed by flow cytometry and microscopic analysis were viewed in HE staining. CCRm-bearing mice were treated with NIH/3T3-LendSN-clone 5 or with NIH/3T3-LXSN cells as a control. ES and G-CSF levels were measured by ELISA and Milliplex, respectively. Quantification of CD11b+Gr-1+ cells and their subsets was performed by flow cytometry. Gr-1+ cells magnetically separated were evaluated in vitro the nitrite levels in supernatant and arginase activity in cells by colorimetric assays. ROS production was measured in CD11b+Gr-1+ cells using the DCFDA marker by flow cytometry. Our data showed the presence of CD11b+Gr-1+ cells in the bone marrow as well as in the kidney, lung and spleen, confirming the monocytic and granulocytic morphologies In metastatic progression, saw the expansion of CD11b+Gr-1+ cells and, preferably, the granulocytic subtype in the bone marrow. Our data demonstrate the progressive accumulation of splenic CD11b+Gr-1+ cells and the opposite was seen in metastatic lungs.Gene therapy with ES reduced the number of pulmonary metastatic lesions, the number of CD11b+Gr-1+ cells and the number of granulocytic cells. G-CSF levels were also reduced and ROS production by granulocytic cells was affected after the treatment with ES. Here, we demonstrate that the metastatic progression is inversely proportional to the number of CD11b+Gr-1+ cells present in the tumor microenvironment, showing that the granulocytic subtype is involved in advanced metastasis. Treatment with ES induced relevant antitumor immune response abrogating the reduction of ROS-producing myeloid-derived suppressor cells in the metastatic local.O carcinoma de células renais (CCR) é a terceira causa de morte de câncer geniturinário. Esse câncer é altamente vascularizado e resistente a radioterapia e quimioterapia. As drogas antiangiogênicas são promissoras e amplamente utilizadas na clínica, porém mecanismo de resistência anti-tumoral tem sido desencadeado com o anticorpo anti-VEGF. Estudos relatam que a resistência ao tratamento de pacientes com câncer está relacionada à presença de células mielóides supressoras. A endostatina (ES) é um fragmento de colágeno XVIII que possui atividade antiangiogênica, porém seus mecanismos de ação não são elucidados. Nesse estudo, rastreamos células Gr-1+ em específicos órgãos do camundongo sadio, avaliamos a importância das células CD11b+Gr-1+ e seus subtipos durante a progressão metastática e examinamos o potencial da terapia com ES na modulação de células CD11b+Gr-1+ e em seus subtipos bem como a atividade imunossupressora na metástase pulmonar. Camundongos Balb/c sadios foram utilizados na detecção de células Gr-1+ separadas magneticamente. Camundongos Balb/c com CCRm foram nefrectomizados no dia 7 e coletados o pulmão metastático, baço e medula óssea após 3, 7 e 10 dias. A quantificação das células CD11b+Gr-1+ e os subtipos, monocítica e granulocítica, foi realizada por citometria de fluxo e análises microscópicas foram visualizadas em coloração de HE. Camundongos Balb/c com CCRm foram tratados com células NIH/3T3-LendSN-clone 5 ou com NIH/3T3-LXSN, como controle. A dosagem de ES foi realizada através de ELISA e do G-CSF através do milliplex. A quantificação de células CD11b+Gr-1+ e os subtipos foi realizada por citometria de fluxo. As células Gr-1+ separadas, magneticamente, foram avaliadas in vitro a dosagem de nitrito no sobrenadante e da atividade de arginase nas células, através de ensaios colorimétricos. E, a produção de EROs foi quantificada em células CD11b+Gr-1+ com o marcador DCFDA por citometria de fluxo. Nossos dados, mostraram a presença de células Gr-1+ tanto na medula óssea quanto no rim, pulmão e baço, confirmando as morfologias mononuclear e polimorfonuclear. No modelo de progressão metastática, verificamos a expansão de células CD11b+Gr-1+ e, preferencialmente, do subtipo granulocítica na medula óssea. Notamos o progressivo acúmulo de células CD11b+Gr-1+ no baço e o oposto nos pulmões metastáticos. A terapia gênica com ES induziu à redução de lesões metastáticas no pulmão, de células CD11b+Gr-1+ e do subtipo granulocítica. A redução dessas células foi refletida na diminuição dos níveis plasmáticos de G-CSF, levando à diminuição da produção de EROs pelas células granulocíticas. Assim, mostramos que a progressão metastática é inversamente proporcional ao número de células CD11b+Gr-1+ presentes no microambiente tumoral, mostrando que o subtipo granulocítica tem participação na metástase avançada. O tratamento com ES induziu uma resposta imune anti-tumoral prejudicando o recrutamento de células mielóides supressoras produtoras de EROs ao sítio metastático.Dados abertos - Sucupira - Teses e dissertações (2013 a 2016

Evaluation of the expression of adhesion molecules (VCAM-1 and ICAM-1) in metastatic renal cell carcinomalung of animals sujected to gene therapy with endostatin

O carcinoma de celulas renais (CCR) e a terceira causa de morte de cancer genitourinario. CCR e elevadamente vascularizado e sensivel a terapia antiangiogenica. Endostatina (ES) e um fragmento de colageno XVIII que possui atividade antiangiogenica. Neste estudo, examinamos o potencial da terapia com ES para ativar celulas endoteliais associadas ao tumor em carcinoma de celulas renais metastatico (CCRm). Camundongos Balb/c com CCRm foram tratados com celulas NIH/3T3-LendSN-clone 3 ou com NIH/3T3-LXSN, como controle. Os subtipos de celulas T no tumor, linfonodo mediastinal e baco foram avaliados por citometria de fluxo. A expressao da molecula de adesao intercelular-1 (ICAM-1) e moleculas de adesao de celula vascular-1 (VCAM-1) foram avaliadas por qPCR em tempo real, citometria de fluxo e analise por imuno-histoquimica. Terapia com ES levou ao aumento na porcentagem de celulas infiltrantes CD4+INF-&#947;+, CD8+INF&#947;+ e CD49b+TNF-&#945;+. Alem disso, ES causou um aumento nos niveis de RNAm de ICAM-1 (1,4 vezes, P<0,01) e VCAM-1 (1,5 vezes) (controle vs ES, P<0,001). Atraves da citometria de fluxo, analisamos um significante aumento das celulas CD34+ICAM-1+ (8,1 vezes, P<0,001) e CD34+VCAM-1+ (1,6 vezes, P<0,05). Terapia genica com ES induziu um significante aumento de ambas as celulas T CD4 e CD8 no linfonodo e baco, sugerindo que ES pode facilitar a sobrevivencia celular ou expansao clonal. Celulas CD49b foram presentes em quantidades aumentadas em todos esses orgaos. Assim, demonstramos que um efeito inflamatorio anti-tumoral de ES em um modelo de CCRm e mediado pelo aumento da expressao de ICAM-1 e VCAM-1 em celulas endoteliais associadas ao tumorBV UNIFESP: Teses e dissertaçõe

Involvement of the NF-kappa B/p50/Bcl-3 complex in response to antiangiogenic therapy in a mouse model of metastatic renal cell carcinoma

Renal cell carcinoma (RCC) represents approximately 2-3% of human malignancies. Nuclear transcription factor kappa B (NF-kappa B) is composed of a family of transcription factors that have been associated with the development and progression of RCC. Endostatin (ES) is a fragment of collagen XVIII that possesses antiangiogenic activity. in this study, we evaluated the expression of NF-kappa B in metastatic tumor cells from animals treated with ES. Balb/c-bearing Renca-EGFP cells were treated with NIH/3T3-LendSN or NIH/3T3-LXSN cells as a control. At the end of the in vivo experiment, plasma Renca-EGFP-sorted cells and tissue lung samples were collected. A real-time PCR array for NF-kappa B target genes revealed that ES therapy led to down regulation of Bcl-3 (P < 0.031), NF-kappa B1 (P < 0.001) and c-Rel (P < 0.004) in the ES-treated group. Using an electrophoretic mobility shift assay (EMSA), we observed a reduction in NF-kappa B binding activity in ES-treated Renca-EGP cells. Furthermore, a supershift assay showed a clear shift of the NF-kappa B DNA band in samples incubated with a p50 antibody. By immunohistochemistry analysis, ES treatment resulted in a significant reduction in expression of p50. (ES vs. control P < 0.05). the immunoprecipitation experiments confirmed the presence of a p50/Bcl-3 complex in nuclear extracts from cells of metastatic lung tissues. Our findings indicate that p50 and Bcl-3 plays a regulatory role in gene transcription in RCC. (C) 2014 Elsevier Masson SAS. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Universidade Federal de São Paulo, Div Nephrol, São Paulo, BrazilUniv São Paulo, Sch Dent, Dept Oral Pathol, São Paulo, BrazilUniv São Paulo, Inst Biomed Sci, Dept Pharmacol, São Paulo, BrazilIPEN CNEN, Dept Biotechnol, São Paulo, BrazilUniversidade Federal de São Paulo, Div Nephrol, São Paulo, BrazilFAPESP: 2010/18969-0CAPES: 72-71859Web of Scienc

Fibronectin expression is decreased in metastatic renal cell carcinoma following endostatin gene therapy

Tumor cells induce the disruption of homeostasis between cellular and extracellular compartments to favor tumor progression. the expression of fibronectin (FN), a matrix glycoprotein, is increased in several carcinoma cell types, including renal cell carcinoma (RCC). RCC are highly vascularized tumors and are often amenable to antiangiogenic therapy. Endostatin (ES) is a fragment of collagen XVIII that possesses antiangiogenic activity. in this study, we examined the modulation of FN gene expression by ES gene therapy in a murine metastatic renal cell carcinoma (mRCC) model. Balb/C mice bearing Renca cells were treated with NIH/3T3-LXSN cells or NIH/3T3-LendSN cells. At the end of the experiment, the ES serum levels were measured, and the FN gene expression was assessed using real-time PCR. the tissue FN was evaluated by western blotting and by immunofluorescence analysis. the ES serum levels in treated mice were higher than those in the control group (P < 0.05). ES treatment led to significant decreases at the FN mRNA (P < 0.001) and protein levels (P < 0.01). Here, we demonstrate the ES antitumor effect that is mediated by down-regulation of FN expression in mRCC. (C) 2012 Elsevier Masson SAS. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Federal de São Paulo, Dept Med, Div Nephrol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biophys, São Paulo, BrazilUniv São Paulo, Sch Vet Med & Anim Sci, Dept Pathol, São Paulo, BrazilUniv São Paulo, Dept Radiol, São Paulo, BrazilUniv São Paulo, IPEN CNEN, Dept Biotechnol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Med, Div Nephrol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biophys, São Paulo, BrazilFAPESP: 2010/18969-0FAPESP: 2009/12518-9Web of Scienc

Fibronectin expression is decreased in metastatic renal cell carcinoma following endostatin gene therapy

Tumor cells induce the disruption of homeostasis between cellular and extracellular compartments to favor tumor progression. The expression of fibronectin (FN), a matrix glycoprotein, is increased in several carcinoma cell types, including renal cell carcinoma (RCC). RCC are highly vascularized tumors and are often amenable to antiangiogenic therapy. Endostatin (ES) is a fragment of collagen XVIII that possesses antiangiogenic activity. In this study, we examined the modulation of FN gene expression by ES gene therapy in a murine metastatic renal cell carcinoma (mRCC) model. Balb/C mice bearing Renca cells were treated with NIH/3T3-LXSN cells or NIH/3T3-LendSN cells. At the end of the experiment, the ES serum levels were measured, and the FN gene expression was assessed using real-time PCR. The tissue FN was evaluated by western blotting and by immunofluorescence analysis. The ES serum levels in treated mice were higher than those in the control group (P &lt; 0.05). ES treatment led to significant decreases at the FN mRNA (P &lt; 0.001) and protein levels (P &lt; 0.01). Here, we demonstrate the ES antitumor effect that is mediated by down-regulation of FN expression in mRCC. (C) 2012 Elsevier Masson SAS. All rights reserved.FAPESP [2010/18969-0, 2009/12518-9]FAPES

Endostatin gene therapy enhances the efficacy of IL-2 in suppressing metastatic renal cell carcinoma in mice

We investigated whether the administration of IL-2 combined with endostatin gene therapy was able to produce additive or even synergistic immunomodulatory activity in a mouse model of metastatic renal carcinoma. Renca cells were injected into the tail vein of BALB/c mice. After 24 h, the animals were randomly divided into four groups (5 mice/group). One group of mice was the control, the second group received treatment with 100,000 UI of Recombinant IL-2 (Proleukin, Chiron) twice a day, 1 day per week during 2 weeks (IL-2), the third group received treatment with a subcutaneous inoculation of 3.6 x 10(6) endostatin-producing cells, and the fourth group received both therapies (IL-2 + ES). Mice were treated for 2 weeks. in the survival studies, 10 mice/group daily, mice were monitored daily until they died. the presence of metastases led to a twofold increase in endostatin levels. Subcutaneous inoculation of NIH/3T3-LendSN cells resulted in a 2.75 and 2.78-fold increase in endostatin levels in the ES and IL-2 + ES group, respectively. At the end of the study, there was a significant decrease in lung wet weight, lung nodules area, and microvascular area (MVA) in all treated groups compared with the control group (P < 0.001). the significant difference in lung wet weight and lung nodules area between groups IL-2 and IL-2 + ES revealed a synergistic antitumor effect of the combined treatment (P < 0.05). the IL-2 + ES therapy Kaplan-Meier survival curves showed that the probability of survival was significantly higher for mice treated with the combined therapy (log-rank test, P = 0.0028). Conjugated therapy caused an increase in the infiltration of CD4, CD8 and CD49b lymphocytes. An increase in the amount of CD8 cells (P < 0.01) was observed when animals received both ES and IL-2, suggesting an additive effect of ES over IL-2 treatment. A synergistic effect of ES on the infiltration of CD4 (P < 0.001) and CD49b cells (P < 0.01) was also observed over the effect of IL-2. Here, we show that ES led to an increase in CD4 T helper cells as well as cytotoxic lymphocytes, such as NK cells and CD8 cells, within tumors of IL-2 treated mice. This means that ES plays a role in supporting the actions of T cells.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Federal de São Paulo, Dept Med, Div Nephrol, São Paulo, BrazilUniv São Paulo, Dept Radiol, São Paulo, BrazilUniv São Paulo, Inst Biomed Sci, Dept Immunol, São Paulo, BrazilAlbert Einstein Jewish Inst Educ & Res, São Paulo, BrazilIPEN CNEN, Dept Biotechnol, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Med, Div Nephrol, São Paulo, BrazilFAPESP: 2007/54253-6Web of Scienc