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Article ; Online: Protocol for designing and bioprinting multi-layered constructs to reconstruct an endothelial-epithelial 3D model.

Backes, Eduardo Henrique / Zamproni, Laura Nicoleti / Delgado-Garcia, Lina Maria / Pinto, Leonardo Alves / Lemes, Robertha Mariana Rodrigues / Bartolomeo, Cynthia Silva / Porcionatto, Marimélia Aparecida

STAR protocols

2023 Volume 4, Issue 3, Page(s) 102467

Abstract: 3D bioprinting has opened new possibilities and elevated tissue engineering complexity. Here, we present a protocol to design a 3D model with two cell lineage layers (A549 and HUVEC) to recreate multi-cell constructs. We describe the steps for slicing ... ...

Abstract	3D bioprinting has opened new possibilities and elevated tissue engineering complexity. Here, we present a protocol to design a 3D model with two cell lineage layers (A549 and HUVEC) to recreate multi-cell constructs. We describe the steps for slicing the constructs, handling hydrogels, and detailing the bioprinting setup. These 3D-bioprinted constructs can be adapted to various cell models-from primary cell cultures to commercial cell lines and induced pluripotent stem cells (IPCs)-and applications, including drug screening and disease modeling. For complete details on the use and execution of this protocol, please refer to Cruz et al.
MeSH term(s)	Bioprinting/methods ; Tissue Engineering/methods ; Hydrogels
Chemical Substances	Hydrogels
Language	English
Publishing date	2023-08-15
Publishing country	United States
Document type	Journal Article
ISSN	2666-1667
ISSN (online)	2666-1667
DOI	10.1016/j.xpro.2023.102467
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Article ; Online: Protocol for designing and bioprinting multi-layered constructs to reconstruct an endothelial-epithelial 3D model

Eduardo Henrique Backes / Laura Nicoleti Zamproni / Lina Maria Delgado-Garcia / Leonardo Alves Pinto / Robertha Mariana Rodrigues Lemes / Cynthia Silva Bartolomeo / Marimélia Aparecida Porcionatto

STAR Protocols, Vol 4, Iss 3, Pp 102467- (2023)

2023

Abstract: Summary: 3D bioprinting has opened new possibilities and elevated tissue engineering complexity. Here, we present a protocol to design a 3D model with two cell lineage layers (A549 and HUVEC) to recreate multi-cell constructs. We describe the steps for ... ...

Abstract	Summary: 3D bioprinting has opened new possibilities and elevated tissue engineering complexity. Here, we present a protocol to design a 3D model with two cell lineage layers (A549 and HUVEC) to recreate multi-cell constructs. We describe the steps for slicing the constructs, handling hydrogels, and detailing the bioprinting setup. These 3D-bioprinted constructs can be adapted to various cell models—from primary cell cultures to commercial cell lines and induced pluripotent stem cells (IPCs)—and applications, including drug screening and disease modeling.For complete details on the use and execution of this protocol, please refer to Cruz et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
Keywords	Cell Culture ; Health Sciences ; Material Sciences ; Tissue Engineering ; Science (General) ; Q1-390
Subject code	571
Language	English
Publishing date	2023-09-01T00:00:00Z
Publisher	Elsevier
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Overexpression of estrogen receptor GPER1 and G1 treatment reduces SARS-CoV-2 infection in BEAS-2B bronchial cells.

Costa, Angelica Jardim / Lemes, Robertha Mariana Rodrigues / Bartolomeo, Cynthia Silva / Nunes, Tamires Alves / Pereira, Gabriela Cruz / Oliveira, Rafaela Brito / Gomes, Alexandre Lopes / Smaili, Soraya Soubhi / Maciel, Rui Monteiro de Barros / Newson, Louise / Ramirez, Ana Lopez / Okuda, Liria Hiromi / Prado, Carla Máximo / Stilhano, Roberta Sessa / Ureshino, Rodrigo Portes

Molecular and cellular endocrinology

2022 Volume 558, Page(s) 111775

Abstract: Gender-bias in COVID-19 severity has been suggested by clinical data. Experimental data in cell and animal models have demonstrated the role of sex hormones, particularly estrogens, in viral infections such as in COVID-19. SARS-CoV-2 uses ACE2 as a ... ...

Abstract	Gender-bias in COVID-19 severity has been suggested by clinical data. Experimental data in cell and animal models have demonstrated the role of sex hormones, particularly estrogens, in viral infections such as in COVID-19. SARS-CoV-2 uses ACE2 as a receptor to recognize host cells, and the protease TMPRSS2 for priming the Spike protein, facilitating virus entry into cells. However, the involvement of estrogenic receptors in SARS-CoV-2 infection are still being explored. Thus, in order to investigate the role of estrogen and its receptors in COVID-19, the estrogen receptors ERα, ERβ and GPER1 were overexpressed in bronchial BEAS-2B cell, and then infected with SARS-CoV-2. Interestingly, the levels of ACE2 and TMPRSS2 mRNA were higher in SARS-CoV-2-infected cells, but no difference was observed in cells with estrogen receptors overexpression. GPER1 can be involved in virus infection or replication, since its higher levels reduces SARS-CoV-2 load. On the other hand, pharmacological antagonism of GPER1 enhanced viral load. Those data suggest that GPER1 has an important role in SARS-CoV-2 infection.
MeSH term(s)	Animals ; COVID-19 ; SARS-CoV-2 ; Angiotensin-Converting Enzyme 2 ; Spike Glycoprotein, Coronavirus/genetics ; Spike Glycoprotein, Coronavirus/metabolism ; Receptors, Estrogen ; Estrogen Receptor beta ; Estrogen Receptor alpha ; Peptidyl-Dipeptidase A/metabolism ; RNA, Messenger/genetics ; Estrogens
Chemical Substances	Angiotensin-Converting Enzyme 2 (EC 3.4.17.23) ; Spike Glycoprotein, Coronavirus ; Receptors, Estrogen ; Estrogen Receptor beta ; Estrogen Receptor alpha ; Peptidyl-Dipeptidase A (EC 3.4.15.1) ; RNA, Messenger ; Estrogens ; spike protein, SARS-CoV-2
Language	English
Publishing date	2022-09-09
Publishing country	Ireland
Document type	Journal Article
ZDB-ID	187438-x
ISSN	1872-8057 ; 0303-7207
ISSN (online)	1872-8057
ISSN	0303-7207
DOI	10.1016/j.mce.2022.111775
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Article ; Online: SARS-CoV-2 infection and replication kinetics in different human cell types: The role of autophagy, cellular metabolism and ACE2 expression.

Bartolomeo, Cynthia Silva / Lemes, Robertha Mariana Rodrigues / Morais, Rafael Leite / Pereria, Gabriela Cruz / Nunes, Tamires Alves / Costa, Angelica Jardim / de Barros Maciel, Rui Monteiro / Braconi, Carla Torres / Maricato, Juliana Terzi / Janini, Luiz Mario Ramos / Okuda, Liria Hiromi / Lee, Kil Sun / Prado, Carla Máximo / Ureshino, Rodrigo Portes / Stilhano, Roberta Sessa

Life sciences

2022 Volume 308, Page(s) 120930

Abstract: Aims: This study evaluated SARS-CoV-2 replication in human cell lines derived from various tissues and investigated molecular mechanisms related to viral infection susceptibility and replication.: Main methods: SARS-CoV-2 replication in BEAS-2B and ... ...

Abstract	Aims: This study evaluated SARS-CoV-2 replication in human cell lines derived from various tissues and investigated molecular mechanisms related to viral infection susceptibility and replication. Main methods: SARS-CoV-2 replication in BEAS-2B and A549 (respiratory tract), HEK-293 T (kidney), HuH7 (liver), SH-SY5Y (brain), MCF7 (breast), Huvec (endothelial) and Caco-2 (intestine) was evaluated by RT-qPCR. Concomitantly, expression levels of ACE2 (Angiotensin Converting Enzyme) and TMPRSS2 were assessed through RT-qPCR and western blot. Proteins related to autophagy and mitochondrial metabolism were monitored in uninfected cells to characterize the cellular metabolism of each cell line. The effect of ACE2 overexpression on viral replication in pulmonary cells was also investigated. Key findings: Our data show that HuH7, Caco-2 and MCF7 presented a higher viral load compared to the other cell lines. The increased susceptibility to SARS-CoV-2 infection seems to be associated not only with the differential levels of proteins intrinsically related to energetic metabolism, such as ATP synthase, citrate synthase, COX and NDUFS2 but also with the considerably higher TMPRSS2 mRNA expression. The two least susceptible cell types, BEAS-2B and A549, showed drastically increased SARS-CoV-2 replication capacity when ACE2 was overexpressed. These modified cell lines are relevant for studying SARS-CoV-2 replication in vitro. Significance: Our data not only reinforce that TMPRSS2 expression and cellular energy metabolism are important molecular mechanisms for SARS-CoV-2 infection and replication, but also indicate that HuH7, MCF7 and Caco-2 are suitable models for mechanistic studies of COVID-19. Moreover, pulmonary cells overexpressing ACE2 can be used to understand mechanisms associated with SARS-CoV-2 replication.
MeSH term(s)	Adenosine Triphosphate ; Angiotensin-Converting Enzyme 2/genetics ; Autophagy ; COVID-19 ; Caco-2 Cells ; Citrate (si)-Synthase ; HEK293 Cells ; Humans ; Neuroblastoma ; Peptidyl-Dipeptidase A/metabolism ; RNA, Messenger/genetics ; SARS-CoV-2
Chemical Substances	RNA, Messenger ; Adenosine Triphosphate (8L70Q75FXE) ; Citrate (si)-Synthase (EC 2.3.3.1) ; Peptidyl-Dipeptidase A (EC 3.4.15.1) ; Angiotensin-Converting Enzyme 2 (EC 3.4.17.23)
Language	English
Publishing date	2022-09-06
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	3378-9
ISSN	1879-0631 ; 0024-3205
ISSN (online)	1879-0631
ISSN	0024-3205
DOI	10.1016/j.lfs.2022.120930
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Article ; Online: Effect of Mycobacterium leprae on neurotrophins expression in human Schwann cells and mouse sciatic nerves.

Nogueira, Maria Renata Sales / Amôr, Nádia Ghinelli / Michellin, Letícia Baccaro / Cury Filho, Milton / Rosa, Patrícia Sammarco / Latini, Ana Carla Pereira / Rodrigues, Luciana Silva / Lemes, Robertha Mariana Rodrigues / Lara, Flavio Alves / Pessolani, Maria Cristina Vidal

Memorias do Instituto Oswaldo Cruz

2020 Volume 115, Page(s) e200075

Abstract: Background: Although Mycobacterium leprae (ML) is well characterised as the causative agent of leprosy, the pathophysiological mechanisms underlying peripheral nerve damage still need further understanding. In vitro and in vivo studies have yielded ... ...

Abstract	Background: Although Mycobacterium leprae (ML) is well characterised as the causative agent of leprosy, the pathophysiological mechanisms underlying peripheral nerve damage still need further understanding. In vitro and in vivo studies have yielded insights into molecular mechanisms of ML interaction with Schwann cells (SC), indicating the regulation of genes and proteins crucial to neural plasticity. Objectives: We aimed to investigate the effect of ML on neurotrophins expression in human SC (hSC) and mice sciatic nerves to better understand their role in leprosy neuropathy, and aiming to contribute to future therapeutic approaches. Methods: We evaluated mRNA and protein expression of BDNF, NGF, NT-3, NT-4 in hSC from amputation nerve fragments, as well as in athymic nude mice, infected by ML for eight months. Findings and main conclusions: Our in vitro results showed a trend to decline in NGF and BDNF mRNA in ML-treated hSC, compared to controls. The immunodetection of BDNF and NT-4 was significantly downregulated in ML-treated hSC. Conversely, ML-infected mice demonstrated upregulation of NT-3, compared to non-infected animals. Our findings indicate that ML may be involved in neurotrophins regulation, suggesting that a pathogen-related imbalance of these growth factors may have a role in the neural impairment of leprosy.
MeSH term(s)	Animals ; Humans ; Mice ; Mice, Nude ; Mycobacterium leprae ; Nerve Growth Factors/metabolism ; Schwann Cells/metabolism ; Sciatic Nerve/metabolism
Chemical Substances	Nerve Growth Factors
Language	English
Publishing date	2020-07-17
Publishing country	Brazil
Document type	Journal Article
ZDB-ID	953293-6
ISSN	1678-8060 ; 0074-0276
ISSN (online)	1678-8060
ISSN	0074-0276
DOI	10.1590/0074-02760200075
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Article: Overexpression of estrogen receptor GPER1 and G1 treatment reduces SARS-CoV-2 infection in BEAS-2B bronchial cells

Molecular and cellular endocrinology. 2022 Dec. 01, v. 558

2022

Abstract	Gender-bias in COVID-19 severity has been suggested by clinical data. Experimental data in cell and animal models have demonstrated the role of sex hormones, particularly estrogens, in viral infections such as in COVID-19. SARS-CoV-2 uses ACE2 as a receptor to recognize host cells, and the protease TMPRSS2 for priming the Spike protein, facilitating virus entry into cells. However, the involvement of estrogenic receptors in SARS-CoV-2 infection are still being explored. Thus, in order to investigate the role of estrogen and its receptors in COVID-19, the estrogen receptors ERα, ERβ and GPER1 were overexpressed in bronchial BEAS-2B cell, and then infected with SARS-CoV-2. Interestingly, the levels of ACE2 and TMPRSS2 mRNA were higher in SARS-CoV-2-infected cells, but no difference was observed in cells with estrogen receptors overexpression. GPER1 can be involved in virus infection or replication, since its higher levels reduces SARS-CoV-2 load. On the other hand, pharmacological antagonism of GPER1 enhanced viral load. Those data suggest that GPER1 has an important role in SARS-CoV-2 infection.
Keywords	COVID-19 infection ; Severe acute respiratory syndrome coronavirus 2 ; animals ; antagonism ; estrogen receptors ; estrogens ; proteinases ; viral load ; viruses
Language	English
Dates of publication	2022-1201
Publishing place	Elsevier B.V.
Document type	Article
ZDB-ID	187438-x
ISSN	1872-8057 ; 0303-7207
ISSN (online)	1872-8057
ISSN	0303-7207
DOI	10.1016/j.mce.2022.111775
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Article ; Online: Effect of Mycobacterium leprae on neurotrophins expression in human Schwann cells and mouse sciatic nerves

Maria Renata Sales Nogueira / Nádia Ghinelli Amôr / Letícia Baccaro Michellin / Milton Cury Filho / Patrícia Sammarco Rosa / Ana Carla Pereira Latini / Luciana Silva Rodrigues / Robertha Mariana Rodrigues Lemes / Flavio Alves Lara / Maria Cristina Vidal Pessolani

Memórias do Instituto Oswaldo Cruz., Vol

2020 Volume 115

Abstract: BACKGROUND Although Mycobacterium leprae (ML) is well characterised as the causative agent of leprosy, the pathophysiological mechanisms underlying peripheral nerve damage still need further understanding. In vitro and in vivo studies have yielded ... ...

Abstract	BACKGROUND Although Mycobacterium leprae (ML) is well characterised as the causative agent of leprosy, the pathophysiological mechanisms underlying peripheral nerve damage still need further understanding. In vitro and in vivo studies have yielded insights into molecular mechanisms of ML interaction with Schwann cells (SC), indicating the regulation of genes and proteins crucial to neural plasticity. OBJECTIVES We aimed to investigate the effect of ML on neurotrophins expression in human SC (hSC) and mice sciatic nerves to better understand their role in leprosy neuropathy, and aiming to contribute to future therapeutic approaches. METHODS We evaluated mRNA and protein expression of BDNF, NGF, NT-3, NT-4 in hSC from amputation nerve fragments, as well as in athymic nude mice, infected by ML for eight months. FINDINGS and MAIN CONCLUSIONS Our in vitro results showed a trend to decline in NGF and BDNF mRNA in ML-treated hSC, compared to controls. The immunodetection of BDNF and NT-4 was significantly downregulated in ML-treated hSC. Conversely, ML-infected mice demonstrated upregulation of NT-3, compared to non-infected animals. Our findings indicate that ML may be involved in neurotrophins regulation, suggesting that a pathogen-related imbalance of these growth factors may have a role in the neural impairment of leprosy.
Keywords	leprosy ; Schwann cells ; neurotrophins ; Arctic medicine. Tropical medicine ; RC955-962 ; Microbiology ; QR1-502
Subject code	570
Language	English
Publishing date	2020-07-01T00:00:00Z
Publisher	Instituto Oswaldo Cruz, Ministério da Saúde
Document type	Article ; Online
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Article: SARS-CoV-2 infection and replication kinetics in different human cell types: The role of autophagy, cellular metabolism and ACE2 expression

Life sciences. 2022 Nov. 01, v. 308

2022

Abstract: This study evaluated SARS-CoV-2 replication in human cell lines derived from various tissues and investigated molecular mechanisms related to viral infection susceptibility and replication. SARS-CoV-2 replication in BEAS-2B and A549 (respiratory tract), ... ...

Abstract	This study evaluated SARS-CoV-2 replication in human cell lines derived from various tissues and investigated molecular mechanisms related to viral infection susceptibility and replication. SARS-CoV-2 replication in BEAS-2B and A549 (respiratory tract), HEK-293 T (kidney), HuH7 (liver), SH-SY5Y (brain), MCF7 (breast), Huvec (endothelial) and Caco-2 (intestine) was evaluated by RT-qPCR. Concomitantly, expression levels of ACE2 (Angiotensin Converting Enzyme) and TMPRSS2 were assessed through RT-qPCR and western blot. Proteins related to autophagy and mitochondrial metabolism were monitored in uninfected cells to characterize the cellular metabolism of each cell line. The effect of ACE2 overexpression on viral replication in pulmonary cells was also investigated. Our data show that HuH7, Caco-2 and MCF7 presented a higher viral load compared to the other cell lines. The increased susceptibility to SARS-CoV-2 infection seems to be associated not only with the differential levels of proteins intrinsically related to energetic metabolism, such as ATP synthase, citrate synthase, COX and NDUFS2 but also with the considerably higher TMPRSS2 mRNA expression. The two least susceptible cell types, BEAS-2B and A549, showed drastically increased SARS-CoV-2 replication capacity when ACE2 was overexpressed. These modified cell lines are relevant for studying SARS-CoV-2 replication in vitro. Our data not only reinforce that TMPRSS2 expression and cellular energy metabolism are important molecular mechanisms for SARS-CoV-2 infection and replication, but also indicate that HuH7, MCF7 and Caco-2 are suitable models for mechanistic studies of COVID-19. Moreover, pulmonary cells overexpressing ACE2 can be used to understand mechanisms associated with SARS-CoV-2 replication.
Keywords	COVID-19 infection ; H-transporting ATP synthase ; Severe acute respiratory syndrome coronavirus 2 ; Western blotting ; autophagy ; brain ; breasts ; cell lines ; citrate (si)-synthase ; energy metabolism ; gene expression ; humans ; intestines ; kidneys ; liver ; mitochondria ; respiratory system ; viral load ; virus replication
Language	English
Dates of publication	2022-1101
Publishing place	Elsevier Inc.
Document type	Article
ZDB-ID	3378-9
ISSN	1879-0631 ; 0024-3205
ISSN (online)	1879-0631
ISSN	0024-3205
DOI	10.1016/j.lfs.2022.120930
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Article ; Online: 17β-estradiol reduces SARS-CoV-2 infection in vitro.

Lemes, Robertha Mariana Rodrigues / Costa, Angelica Jardim / Bartolomeo, Cynthia Silva / Bassani, Taysa Bervian / Nishino, Michelle Sayuri / Pereira, Gustavo Jose da Silva / Smaili, Soraya Soubhi / Maciel, Rui Monteiro de Barros / Braconi, Carla Torres / da Cruz, Edgar Ferreira / Ramirez, Ana Lopez / Maricatto, Juliana Terzi / Janini, Luiz Mario Ramos / Prado, Carla Máximo / Stilhano, Roberta Sessa / Ureshino, Rodrigo Portes

Physiological reports

2021 Volume 9, Issue 2, Page(s) e14707

Abstract: The COVID-19 has originated from Wuhan, China, in December 2019 and has been affecting the public health system, society, and economy in an unheard-of manner. There is no specific treatment or vaccine available for COVID-19. Previous data showed that men ...

Abstract	The COVID-19 has originated from Wuhan, China, in December 2019 and has been affecting the public health system, society, and economy in an unheard-of manner. There is no specific treatment or vaccine available for COVID-19. Previous data showed that men are more affected than women by COVID-19, then we hypothesized whether sex hormones could be protecting the female organism against the infection. VERO E6 cells have been commonly used as in vitro model for SARS-CoV-2 infection. In our experimental approach, we have treated VERO E6 cells with 17β-estradiol to evaluate the modulation of SARS-CoV-2 infection in this cell line. Here we demonstrated that estrogen protein receptors ERα, ERβ, and GPER1 are expressed by VERO E6 cells and could be used to study the effects of this steroid hormone. Previous and 24-hours post-infection, cells treated with 17β-estradiol revealed a reduction in the viral load. Afterward, we found that SARS-CoV-2 infection per se results in ACE2 and TMPRSS2 increased gene expression in VERO E6-cell, which could be generating a cycle of virus infection in host cells. The estrogen treatment reduces the levels of the TMPRSS2, which are involved with SARS-CoV-2 infectiveness capacity, and hence, reducing the pathogenicity/genesis. These data suggest that estrogen could be a potential therapeutic target promoting cell protection against SARS-CoV-2. This opens new possibilities for further studies on 17β-estradiol in human cell lines infected by SARS-CoV-2 and at least in part, explain why men developed a more severe COVID-19 compared to women.
MeSH term(s)	Angiotensin-Converting Enzyme 2/genetics ; Angiotensin-Converting Enzyme 2/metabolism ; Animals ; Antiviral Agents/pharmacology ; COVID-19/metabolism ; COVID-19/virology ; Chlorocebus aethiops ; Estradiol/pharmacology ; Host-Pathogen Interactions ; Receptors, Virus/genetics ; Receptors, Virus/metabolism ; SARS-CoV-2/drug effects ; SARS-CoV-2/pathogenicity ; Serine Endopeptidases/genetics ; Serine Endopeptidases/metabolism ; Vero Cells ; COVID-19 Drug Treatment
Chemical Substances	Antiviral Agents ; Receptors, Virus ; Estradiol (4TI98Z838E) ; Angiotensin-Converting Enzyme 2 (EC 3.4.17.23) ; Serine Endopeptidases (EC 3.4.21.-)
Language	English
Publishing date	2021-01-18
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2724325-4
ISSN	2051-817X ; 2051-817X
ISSN (online)	2051-817X
ISSN	2051-817X
DOI	10.14814/phy2.14707
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Article ; Online: STING-Dependent 2'-5' Oligoadenylate Synthetase-Like Production Is Required for Intracellular Mycobacterium leprae Survival.

de Toledo-Pinto, Thiago Gomes / Ferreira, Anna Beatriz Robottom / Ribeiro-Alves, Marcelo / Rodrigues, Luciana Silva / Batista-Silva, Leonardo Ribeiro / Silva, Bruno Jorge de Andrade / Lemes, Robertha Mariana Rodrigues / Martinez, Alejandra Nóbrega / Sandoval, Felipe Galvan / Alvarado-Arnez, Lucia Elena / Rosa, Patrícia Sammarco / Shannon, Edward Joseph / Pessolani, Maria Cristina Vidal / Pinheiro, Roberta Olmo / Antunes, Sérgio Luís Gomes / Sarno, Euzenir Nunes / Lara, Flávio Alves / Williams, Diana Lynn / Ozório Moraes, Milton

The Journal of infectious diseases

2016 Volume 214, Issue 2, Page(s) 311–320

Abstract: Cytosolic detection of nucleic acids elicits a type I interferon (IFN) response and plays a critical role in host defense against intracellular pathogens. Herein, a global gene expression profile of Mycobacterium leprae-infected primary human Schwann ... ...

Abstract	Cytosolic detection of nucleic acids elicits a type I interferon (IFN) response and plays a critical role in host defense against intracellular pathogens. Herein, a global gene expression profile of Mycobacterium leprae-infected primary human Schwann cells identified the genes differentially expressed in the type I IFN pathway. Among them, the gene encoding 2'-5' oligoadenylate synthetase-like (OASL) underwent the greatest upregulation and was also shown to be upregulated in M. leprae-infected human macrophage cell lineages, primary monocytes, and skin lesion specimens from patients with a disseminated form of leprosy. OASL knock down was associated with decreased viability of M. leprae that was concomitant with upregulation of either antimicrobial peptide expression or autophagy levels. Downregulation of MCP-1/CCL2 release was also observed during OASL knock down. M. leprae-mediated OASL expression was dependent on cytosolic DNA sensing mediated by stimulator of IFN genes signaling. The addition of M. leprae DNA enhanced nonpathogenic Mycobacterium bovis bacillus Calmette-Guerin intracellular survival, downregulated antimicrobial peptide expression, and increased MCP-1/CCL2 secretion. Thus, our data uncover a promycobacterial role for OASL during M. leprae infection that directs the host immune response toward a niche that permits survival of the pathogen.
MeSH term(s)	2',5'-Oligoadenylate Synthetase/metabolism ; Cells, Cultured ; Epithelial Cells/microbiology ; Gene Expression Profiling ; Gene Knockdown Techniques ; Host-Pathogen Interactions ; Humans ; Leprosy/microbiology ; Leprosy/pathology ; Macrophages/microbiology ; Membrane Proteins/metabolism ; Microbial Viability ; Mycobacterium bovis/physiology ; Mycobacterium leprae/physiology ; Schwann Cells/microbiology
Chemical Substances	Membrane Proteins ; STING1 protein, human ; OASL protein, human (EC 2.7.7.-) ; 2',5'-Oligoadenylate Synthetase (EC 2.7.7.84)
Language	English
Publishing date	2016-05-14
Publishing country	United States
Document type	Journal Article
ZDB-ID	3019-3
ISSN	1537-6613 ; 0022-1899
ISSN (online)	1537-6613
ISSN	0022-1899
DOI	10.1093/infdis/jiw144
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