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  1. Article ; Online: Trained innate immunity, COVID-19 therapeutic dilemma, and fake science.

    Belizário, José Ernesto

    Clinics (Sao Paulo, Brazil)

    2020  Volume 75, Page(s) e2124

    MeSH term(s) BCG Vaccine/therapeutic use ; Betacoronavirus ; COVID-19 ; Chloroquine/therapeutic use ; Clinical Trials as Topic ; Coronavirus Infections/immunology ; Coronavirus Infections/therapy ; Humans ; Immunity, Innate ; Pandemics ; Pneumonia, Viral/immunology ; Pneumonia, Viral/therapy ; SARS-CoV-2 ; Vaccination
    Chemical Substances BCG Vaccine ; Chloroquine (886U3H6UFF)
    Keywords covid19
    Language English
    Publishing date 2020-07-10
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2182801-5
    ISSN 1980-5322 ; 1807-5932
    ISSN (online) 1980-5322
    ISSN 1807-5932
    DOI 10.6061/clinics/2020/e2124
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Book ; Online: Trained innate immunity, COVID-19 therapeutic dilemma, and fake science

    Belizário, José Ernesto

    Clinics v.75 2020

    2020  

    Keywords covid19
    Language English
    Publishing date 2020-01-01
    Publisher Faculdade de Medicina / USP
    Publishing country br
    Document type Book ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  4. Article ; Online: Trained innate immunity, COVID-19 therapeutic dilemma, and fake science

    José Ernesto Belizário

    Clinics, Vol

    2020  Volume 75

    Keywords Medicine (General) ; R5-920
    Language English
    Publishing date 2020-07-01T00:00:00Z
    Publisher Elsevier España
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  5. Article ; Online: Trained innate immunity, COVID-19 therapeutic dilemma, and fake science

    Belizário, José Ernesto

    Clinics; v.; e2124 ; Clinics; Vol. 75 (2020); e2124 ; Clinics; Vol 75 (2020); e2124 ; 1980-5322 ; 1807-5932

    2020  Volume 75

    Keywords covid19
    Language English
    Publishing date 2020-07-31
    Publisher Hospital das Clínicas, Faculdade de Medicina, Universidade de São Paulo
    Publishing country br
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  6. Article ; Online: Characterization of post-edited cells modified in the TFAM gene by CRISPR/Cas9 technology in the bovine model.

    de Oliveira, Vanessa Cristina / Gomes Mariano Junior, Clésio / Belizário, José Ernesto / Krieger, José Eduardo / Fernandes Bressan, Fabiana / Roballo, Kelly Cristine Santos / Fantinato-Neto, Paulo / Meirelles, Flávio Vieira / Chiaratti, Marcos Roberto / Concordet, Jean-Paul / Ambrósio, Carlos Eduardo

    PloS one

    2020  Volume 15, Issue 7, Page(s) e0235856

    Abstract: Gene editing in large animal models for future applications in translational medicine and food production must be deeply investigated for an increase of knowledge. The mitochondrial transcription factor A (TFAM) is a member of the HMGB subfamily that ... ...

    Abstract Gene editing in large animal models for future applications in translational medicine and food production must be deeply investigated for an increase of knowledge. The mitochondrial transcription factor A (TFAM) is a member of the HMGB subfamily that binds to mtDNA promoters. This gene maintains mtDNA, and it is essential for the initiation of mtDNA transcription. Lately, we generated a new cell line through the disruption of the TFAM gene in bovine fibroblast cells by CRISPR/Cas 9 technology. We showed that the CRISPR/Cas9 design was efficient through the generation of heterozygous mutant clones. In this context, once this gene regulates the mtDNA replication specificity, the study aimed to determine if the post-edited cells are capable of in vitro maintenance and assess if they present changes in mtDNA copies and mitochondrial membrane potential after successive passages in culture. The post-edited cells were expanded in culture, and we performed a growth curve, doubling time, cell viability, mitochondrial DNA copy number, and mitochondrial membrane potential assays. The editing process did not make cell culture unfeasible, even though cell growth rate and viability were decreased compared to control since we observed the cells grow well when cultured in a medium supplemented with uridine and pyruvate. They also exhibited a classical fibroblastoid appearance. The RT-qPCR to determine the mtDNA copy number showed a decrease in the edited clones compared to the non-edited ones (control) in different cell passages. Cell staining with Mitotracker Green and red suggests a reduction in red fluorescence in the edited cells compared to the non-edited cells. Thus, through characterization, we demonstrated that the TFAM gene is critical to mitochondrial maintenance due to its interference in the stability of the mitochondrial DNA copy number in different cell passages and membrane potential confirming the decrease in mitochondrial activity in cells edited in heterozygosis.
    MeSH term(s) Animals ; CRISPR-Cas Systems ; Cattle/genetics ; Cells, Cultured ; DNA Replication ; DNA, Mitochondrial/genetics ; DNA-Binding Proteins/genetics ; Fibroblasts/metabolism ; Gene Dosage ; Gene Editing ; Mitochondria/genetics ; Mitochondrial Proteins/genetics ; Transcription Factors/genetics
    Chemical Substances DNA, Mitochondrial ; DNA-Binding Proteins ; Mitochondrial Proteins ; Transcription Factors ; mitochondrial transcription factor A
    Language English
    Publishing date 2020-07-10
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 1932-6203
    ISSN (online) 1932-6203
    DOI 10.1371/journal.pone.0235856
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article ; Online: Characterization of post-edited cells modified in the TFAM gene by CRISPR/Cas9 technology in the bovine model.

    Vanessa Cristina de Oliveira / Clésio Gomes Mariano Junior / José Ernesto Belizário / José Eduardo Krieger / Fabiana Fernandes Bressan / Kelly Cristine Santos Roballo / Paulo Fantinato-Neto / Flávio Vieira Meirelles / Marcos Roberto Chiaratti / Jean-Paul Concordet / Carlos Eduardo Ambrósio

    PLoS ONE, Vol 15, Iss 7, p e

    2020  Volume 0235856

    Abstract: Gene editing in large animal models for future applications in translational medicine and food production must be deeply investigated for an increase of knowledge. The mitochondrial transcription factor A (TFAM) is a member of the HMGB subfamily that ... ...

    Abstract Gene editing in large animal models for future applications in translational medicine and food production must be deeply investigated for an increase of knowledge. The mitochondrial transcription factor A (TFAM) is a member of the HMGB subfamily that binds to mtDNA promoters. This gene maintains mtDNA, and it is essential for the initiation of mtDNA transcription. Lately, we generated a new cell line through the disruption of the TFAM gene in bovine fibroblast cells by CRISPR/Cas 9 technology. We showed that the CRISPR/Cas9 design was efficient through the generation of heterozygous mutant clones. In this context, once this gene regulates the mtDNA replication specificity, the study aimed to determine if the post-edited cells are capable of in vitro maintenance and assess if they present changes in mtDNA copies and mitochondrial membrane potential after successive passages in culture. The post-edited cells were expanded in culture, and we performed a growth curve, doubling time, cell viability, mitochondrial DNA copy number, and mitochondrial membrane potential assays. The editing process did not make cell culture unfeasible, even though cell growth rate and viability were decreased compared to control since we observed the cells grow well when cultured in a medium supplemented with uridine and pyruvate. They also exhibited a classical fibroblastoid appearance. The RT-qPCR to determine the mtDNA copy number showed a decrease in the edited clones compared to the non-edited ones (control) in different cell passages. Cell staining with Mitotracker Green and red suggests a reduction in red fluorescence in the edited cells compared to the non-edited cells. Thus, through characterization, we demonstrated that the TFAM gene is critical to mitochondrial maintenance due to its interference in the stability of the mitochondrial DNA copy number in different cell passages and membrane potential confirming the decrease in mitochondrial activity in cells edited in heterozygosis.
    Keywords Medicine ; R ; Science ; Q
    Subject code 570
    Language English
    Publishing date 2020-01-01T00:00:00Z
    Publisher Public Library of Science (PLoS)
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  8. Article ; Online: FAM3B/PANDER inhibits cell death and increases prostate tumor growth by modulating the expression of Bcl-2 and Bcl-X

    Maciel-Silva, Paula / Caldeira, Izabela / de Assis Santos, Icaro / Carreira, Ana Claudia Oliveira / Siqueira, Flavia Ramos / Antonioli, Eliane / Goldberg, Anna Carla / Belizário, José Ernesto / Garay-Malpartida, Humberto Miguel

    BMC cancer

    2018  Volume 18, Issue 1, Page(s) 90

    Abstract: Background: FAM3B/PANDER is a novel cytokine-like protein that induces apoptosis in insulin-secreting beta-cells. Since in silico data revealed that FAM3B can be expressed in prostate tumors, we evaluated the putative role of this cytokine in prostate ... ...

    Abstract Background: FAM3B/PANDER is a novel cytokine-like protein that induces apoptosis in insulin-secreting beta-cells. Since in silico data revealed that FAM3B can be expressed in prostate tumors, we evaluated the putative role of this cytokine in prostate tumor progression.
    Methods: FAM3B expression was analyzed by quantitative PCR in tumor tissue clinical samples and prostate tumor cell lines. Culture growth and viability of DU145 cell line were evaluated after treatment with either exogenous FAM3B protein obtained from conditioned media (CM) of 293 T cells overexpressing FAM3B or a recombinant FAM3B protein produced in a bacterial host. DU145 cells overexpressing FAM3B protein were produced by lentiviral-mediated transduction of full-length FAM3B cDNA. Cell viability and apoptosis were analyzed in DU145/FAM3B cells after treatment with several cell death inducers, such as TNF-alpha, staurosporine, etoposide, camptothecin, and serum starvation conditions. Anchorage-independent growth in soft agarose assay was used to evaluate in vitro tumorigenicity. In vivo tumorigenicity and invasiveness were evaluated by tumor xenograft growth in nude mice.
    Results: We observed an increase in FAM3B expression in prostate tumor samples when compared to normal tissues. DU145 cell viability and survival increased after exogenous treatment with recombinant FAM3B protein or FAM3B-secreted protein. Overexpression of FAM3B in DU145 cells promoted inhibition of DNA fragmentation and phosphatidylserine externalization in a time and dose-dependent fashion, upon apoptosis triggered by TNF-alpha. These events were accompanied by increased gene expression of anti-apoptotic Bcl-2 and Bcl-XL, decreased expression of pro-apoptotic Bax and diminished caspase-3, -8 and -9 proteolytic activities. Furthermore, inhibition of Bcl-2 anti-apoptotic family proteins with small molecules antagonists decreases protective effects of FAM3B in DU145 cells. When compared to the respective controls, cells overexpressing FAM3B displayed a decreased anchorage- independent growth in vitro and increased tumor growth in xenografted nude mice. The immunohistochemistry analysis of tumor xenografts revealed a similar anti-apoptotic phenotype displayed by FAM3B-overexpressing tumor cells.
    Conclusions: Taken together, by activating pro-survival mechanisms FAM3B overexpression contributes to increased resistance to cell death and tumor growth in nude mice, highlighting a putative role for this cytokine in prostate cancer progression.
    MeSH term(s) Animals ; Apoptosis/drug effects ; Apoptosis/genetics ; Biomarkers, Tumor/genetics ; Camptothecin/administration & dosage ; Cell Death/drug effects ; Cell Line, Tumor ; Cell Survival/genetics ; Cytokines/genetics ; Humans ; Male ; Mice ; Neoplasm Proteins/genetics ; Prostate/metabolism ; Prostatic Neoplasms/drug therapy ; Prostatic Neoplasms/genetics ; Prostatic Neoplasms/pathology ; Proto-Oncogene Proteins c-bcl-2/genetics ; Tumor Necrosis Factor-alpha/genetics ; Xenograft Model Antitumor Assays ; bcl-X Protein/genetics
    Chemical Substances BCL2 protein, human ; BCL2L1 protein, human ; Biomarkers, Tumor ; Cytokines ; FAM3B protein, human ; Neoplasm Proteins ; Proto-Oncogene Proteins c-bcl-2 ; Tumor Necrosis Factor-alpha ; bcl-X Protein ; Camptothecin (XT3Z54Z28A)
    Language English
    Publishing date 2018--22
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 1471-2407
    ISSN (online) 1471-2407
    DOI 10.1186/s12885-017-3950-9
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  9. Article ; Online: Role of SOCS-1 Gene on Melanoma Cell Growth and Tumor Development.

    Scutti, Jorge A Borin / Matsuo, Alisson Leonardo / Pereira, Felipe Valença / Massaoka, Mariana Hiromi / Figueiredo, Carlos Rogério / Moreira, Dayson Friaça / Belizário, José Ernesto / Travassos, Luiz R

    Translational oncology

    2011  Volume 4, Issue 2, Page(s) 101–109

    Abstract: Melanoma is the most aggressive form of skin cancer, and its incidence has increased dramatically over the years. The murine B16F10 melanoma in syngeneic C57Bl/6 mice has been used as a highly aggressive model to investigate tumor development. Presently, ...

    Abstract Melanoma is the most aggressive form of skin cancer, and its incidence has increased dramatically over the years. The murine B16F10 melanoma in syngeneic C57Bl/6 mice has been used as a highly aggressive model to investigate tumor development. Presently, we demonstrate in the B16F10-Nex2 subclone that silencing of SOCS-1, a negative regulator of Jak/Stat pathway, leads to reversal of the tumorigenic phenotype and inhibition of melanoma cell metastasis. SOCS-1 silencing with short hairpin RNA affected tumor growth and cell cycle regulation with arrest at the S phase with large-sized nuclei, reduced cell motility, and decreased melanoma cell invasion through Matrigel. A clonogenic assay showed that SOCS-1 acted as a modulator of resistance to anoikis. In addition, downregulation of SOCS-1 decreased the expression of epidermal growth factor receptor (mainly the phosphorylated-R), Ins-Rα, and fibroblast growth factor receptor. In vivo, silencing of SOCS-1 inhibited subcutaneous tumor growth and metastatic development in the lungs. Because SOCS-1 is expressed in most melanoma cell lines and bears a relation with tumor invasion, thickness, and stage of disease, the present results on the effects of SOCS-1 silencing in melanoma suggest that this regulating protein can be a target of cancer therapy.
    Language English
    Publishing date 2011-04-01
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2443840-6
    ISSN 1936-5233 ; 1936-5233
    ISSN (online) 1936-5233
    ISSN 1936-5233
    DOI 10.1593/tlo.10250
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  10. Article: Expression of the selectable marker gene bsrm in BALB/MK cells induces apoptosis by overproduction of hydrogen peroxide.

    Takeshita, Daniela / Bento, Fernanda Mara / Chammas, Roger / Belizário, José Ernesto / Carmona, Adriana Karaoglanovic / Konno, Katsuhiro / Lopes de Melo, Robson / Molina, Gustavo / Lisboa, Bianca Cristina Garcia / Han, Sang Won

    Biochemistry and cell biology = Biochimie et biologie cellulaire

    2007  Volume 85, Issue 5, Page(s) 573–581

    Abstract: Transduction of the retroviral vector LBmSN, which expresses the blasticidin S resistance gene bsrm in the murine keratinocyte cell line BALB/MK, induces death in these cells. Cell death is caused by a factor called DOKEB (death factor obtained from ... ...

    Abstract Transduction of the retroviral vector LBmSN, which expresses the blasticidin S resistance gene bsrm in the murine keratinocyte cell line BALB/MK, induces death in these cells. Cell death is caused by a factor called DOKEB (death factor obtained from keratinocytes expressing bsrm), which is released before the cells' death. In this report we describe and discuss the purification and characterization of DOKEB. Our results were as follows. (i) The 5-day-old medium from the modified BALB/MK cells with LBmSN was used for purification and characterization by filtration and chromatography: DOKEB was a stable and highly hydrophilic compound, with a molecular mass less than that of 1 amino acid. (ii) The conditioned medium containing DOKEB was reactive against thiobarbituric acid and dichlorofluorescein diacetate. (iii) DOKEB activity was neutralized by the incubation of the conditioned medium with catalase. Therefore, our conclusion is that the BALB/MK cells expressing bsrm produce a large amount of hydrogen peroxide, which catalyzes the process of apoptosis of those cells.
    MeSH term(s) Aminohydrolases/genetics ; Aminohydrolases/metabolism ; Animals ; Apoptosis ; Apoptosis Regulatory Proteins/isolation & purification ; Apoptosis Regulatory Proteins/metabolism ; Cell Death ; Cell Line ; Drug Resistance/genetics ; Genetic Markers ; Genetic Vectors/genetics ; Hydrogen Peroxide/metabolism ; Keratinocytes/metabolism ; Mice ; Mice, Inbred BALB C ; Retroviridae/genetics ; Transduction, Genetic
    Chemical Substances Apoptosis Regulatory Proteins ; Genetic Markers ; Hydrogen Peroxide (BBX060AN9V) ; Aminohydrolases (EC 3.5.4.-) ; blasticidin S deaminase (EC 3.5.4.23)
    Language English
    Publishing date 2007-10
    Publishing country Canada
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 54104-7
    ISSN 0829-8211
    ISSN 0829-8211
    DOI 10.1139/o07-058
    Database MEDical Literature Analysis and Retrieval System OnLINE

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