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  1. Article ; Online: Short-Term Functional and Morphological Changes in the Primary Cultures of Trigeminal Ganglion Cells.

    Veríssimo, Carla Pires / Acosta Filha, Lionete Gall / Moreira da Silva, Fábio Jorge / Westgarth, Harrison / Coelho Aguiar, Juliana De Mattos / Pontes, Bruno / Moura-Neto, Vivaldo / Gazerani, Parisa / DosSantos, Marcos F

    Current issues in molecular biology

    2022  Volume 44, Issue 3, Page(s) 1257–1272

    Abstract: Several studies have proved that glial cells, as well as neurons, play a role in pain pathophysiology. Most of these studies have focused on the contribution of central glial cells (e.g., microglia and astrocytes) to neuropathic pain. Likewise, some ... ...

    Abstract Several studies have proved that glial cells, as well as neurons, play a role in pain pathophysiology. Most of these studies have focused on the contribution of central glial cells (e.g., microglia and astrocytes) to neuropathic pain. Likewise, some works have suggested that peripheral glial cells, particularly satellite glial cells (SGCs), and the crosstalk between these cells and the sensory neurons located in the peripheral ganglia, play a role in the phenomenon that leads to pain. Nonetheless, the study of SGCs may be challenging, as the validity of studying those cells in vitro is still controversial. In this study, a research protocol was developed to examine the potential use of primary mixed neuronal-glia cell cultures obtained from the trigeminal ganglion cells (TGCs) of neonate mice (P10-P12). Primary cultures were established and analyzed at 4 h, 24 h, and 48 h. To this purpose, phase contrast microscopy, immunocytochemistry with antibodies against anti-βIII-tubulin and Sk3, scanning electron microscopy, and time-lapse photography were used. The results indicated the presence of morphological changes in the cultured SGCs obtained from the TGCs. The SGCs exhibited a close relationship with neurons. They presented a round shape in the first 4 h, and a more fusiform shape at 24 h and 48 h of culture. On the other hand, neurons changed from a round shape to a more ramified shape from 4 h to 48 h. Intriguingly, the expression of SK3, a marker of the SGCs, was high in all samples at 4 h, with some cells double-staining for SK3 and βIII-tubulin. The expression of SK3 decreased at 24 h and increased again at 48 h in vitro. These results confirm the high plasticity that the SGCs may acquire in vitro. In this scenario, the authors hypothesize that, at 4 h, a group of the analyzed cells remained undifferentiated and, therefore, were double-stained for SK3 and βIII-tubulin. After 24 h, these cells started to differentiate into SCGs, which was clearer at 48 h in the culture. Mixed neuronal-glial TGC cultures might be implemented as a platform to study the plasticity and crosstalk between primary sensory neurons and SGCs, as well as its implications in the development of chronic orofacial pain.
    Language English
    Publishing date 2022-03-08
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2000024-8
    ISSN 1467-3045 ; 1467-3037
    ISSN (online) 1467-3045
    ISSN 1467-3037
    DOI 10.3390/cimb44030084
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Cyanovirin-N binds to select SARS-CoV-2 spike oligosaccharides outside of the receptor binding domain and blocks infection by SARS-CoV-2.

    Muñoz-Basagoiti, Jordana / Monteiro, Fábio Luís Lima / Krumpe, Lauren R H / Armario-Najera, Victoria / Shenoy, Shilpa R / Perez-Zsolt, Daniel / Westgarth, Harrison James / Villorbina, Gemma / Bomfim, Larissa Maciel / Raïch-Regué, Dàlia / Nogueras, Lara / Henrich, Curtis J / Gallemí, Marçal / Moreira, Filipe Romero Rebello / Torres, Pascual / Wilson, Jennifer / D'arc, Mirela / Marfil, Silvia / Herlinger, Alice Laschuk /
    Pradenas, Edwards / Higa, Luiza Mendonça / Portero-Otin, Manuel / Trinité, Benjamin / Twyman, Richard M / Capell, Teresa / Tanuri, Amilcar / Blanco, Julià / Izquierdo-Useros, Nuria / Rech, Elibio L / Christou, Paul / O'Keefe, Barry R

    Proceedings of the National Academy of Sciences of the United States of America

    2023  Volume 120, Issue 10, Page(s) e2214561120

    Abstract: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an enveloped positive stranded RNA virus which has caused the recent deadly pandemic called COVID-19. The SARS-CoV-2 virion is coated with a heavily glycosylated Spike glycoprotein which is ... ...

    Abstract Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an enveloped positive stranded RNA virus which has caused the recent deadly pandemic called COVID-19. The SARS-CoV-2 virion is coated with a heavily glycosylated Spike glycoprotein which is responsible for attachment and entry into target cells. One, as yet unexploited strategy for preventing SARS-CoV-2 infections, is the targeting of the glycans on Spike. Lectins are carbohydrate-binding proteins produced by plants, algae, and cyanobacteria. Some lectins can neutralize enveloped viruses displaying external glycoproteins, offering an alternative therapeutic approach for the prevention of infection with virulent β-coronaviruses, such as SARS-CoV-2. Here we show that the cyanobacterial lectin cyanovirin-N (CV-N) can selectively target SARS-CoV-2 Spike oligosaccharides and inhibit SARS-CoV-2 infection in vitro and in vivo. CV-N neutralizes Delta and Omicron variants in vitro better than earlier circulating viral variants. CV-N binds selectively to Spike with a Kd as low as 15 nM and a stoichiometry of 2 CV-N: 1 Spike but does not bind to the receptor binding domain (RBD). Further mapping of CV-N binding sites on Spike shows that select high-mannose oligosaccharides in the S1 domain of Spike are targeted by CV-N. CV-N also reduced viral loads in the nares and lungs in vivo to protect hamsters against a lethal viral challenge. In summary, we present an anti-coronavirus agent that works by an unexploited mechanism and prevents infection by a broad range of SARS-CoV-2 strains.
    MeSH term(s) Animals ; Cricetinae ; SARS-CoV-2 ; COVID-19 ; Oligosaccharides/pharmacology ; Lectins
    Chemical Substances cyanovirin N (184539-38-6) ; Oligosaccharides ; Lectins
    Language English
    Publishing date 2023-02-28
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 209104-5
    ISSN 1091-6490 ; 0027-8424
    ISSN (online) 1091-6490
    ISSN 0027-8424
    DOI 10.1073/pnas.2214561120
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Microglia activation in Niemann-Pick disease, type C1 is amendable to therapeutic intervention.

    Cougnoux, Antony / Drummond, Rebecca A / Collar, Amanda L / Iben, James R / Salman, Alexander / Westgarth, Harrison / Wassif, Christopher A / Cawley, Niamh X / Farhat, Nicole Y / Ozato, Keiko / Lionakis, Michail S / Porter, Forbes D

    Human molecular genetics

    2018  Volume 27, Issue 12, Page(s) 2076–2089

    Abstract: Niemann-Pick disease, type C1 (NPC1) is a neurodegenerative disorder with limited treatment options. NPC1 is associated with neuroinflammation; however, attempts to therapeutically target neuroinflammation in NPC1 have had mixed success. We show here ... ...

    Abstract Niemann-Pick disease, type C1 (NPC1) is a neurodegenerative disorder with limited treatment options. NPC1 is associated with neuroinflammation; however, attempts to therapeutically target neuroinflammation in NPC1 have had mixed success. We show here that NPC1 neuroinflammation is characterized by an atypical microglia activation phenotype. Specifically, Npc1-/- microglia demonstrated altered morphology, reduced levels of lineage markers and a shift toward glycolytic metabolism. Treatment with 2-hydroxypropyl-β-cyclodextrin (HPβCD), a drug currently being studied in a phase 2b/3 clinical trial, reversed all microglia-associated defects in Npc1-/- animals. In addition, impairing microglia mediated neuroinflammation by genetic deletion of IRF8 led to decreased symptoms and increased lifespan. We identified CD22 as a marker of dysregulated microglia in Npc1 mutant mice and subsequently demonstrated that elevated cerebrospinal fluid levels of CD22 in NPC1 patients responds to HPβCD administration. Collectively, these data provide the first in-depth analysis of microglia function in NPC1 and suggest possible new therapeutic approaches.
    MeSH term(s) 2-Hydroxypropyl-beta-cyclodextrin/administration & dosage ; Adolescent ; Adult ; Animals ; Cerebellum/drug effects ; Cerebellum/pathology ; Child ; Child, Preschool ; Disease Models, Animal ; Female ; Humans ; Infant ; Inflammation/drug therapy ; Inflammation/genetics ; Inflammation/pathology ; Interferon Regulatory Factors ; Male ; Mice ; Microglia/drug effects ; Microglia/pathology ; Niemann-Pick C1 Protein/genetics ; Niemann-Pick Disease, Type C/cerebrospinal fluid ; Niemann-Pick Disease, Type C/drug therapy ; Niemann-Pick Disease, Type C/genetics ; Niemann-Pick Disease, Type C/pathology ; Sialic Acid Binding Ig-like Lectin 2/cerebrospinal fluid ; Sialic Acid Binding Ig-like Lectin 2/genetics
    Chemical Substances Interferon Regulatory Factors ; Niemann-Pick C1 Protein ; Sialic Acid Binding Ig-like Lectin 2 ; interferon regulatory factor-8 ; 2-Hydroxypropyl-beta-cyclodextrin (1I96OHX6EK)
    Language English
    Publishing date 2018-03-31
    Publishing country England
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, N.I.H., Intramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 1108742-0
    ISSN 1460-2083 ; 0964-6906
    ISSN (online) 1460-2083
    ISSN 0964-6906
    DOI 10.1093/hmg/ddy112
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Identification and characterisation of SARS-CoV-2 and Human alphaherpesvirus 1 from a productive coinfection in a fatal COVID-19 case.

    Herlinger, Alice Laschuk / Monteiro, Fábio Luís Lima / D'arc, Mirela / Moreira, Filipe Romero Rebello / Westgarth, Harrison James / Galliez, Rafael Mello / Mariani, Diana / da Costa, Luciana Jesus / de Almeida, Luiz Gonzaga Paula / Voloch, Carolina Moreira / Melo, Adriana Suely de Oliveira / Aguiar, Renato Santana de / Dos Santos, André Felipe Andrade / Castiñeiras, Terezinha Marta Pereira Pinto / de Vasconcelos, Ana Tereza Ribeiro / João Filho, Esaú Custódio / Escosteguy, Claudia Caminha / Ferreira Junior, Orlando da Costa / Tanuri, Amilcar /
    Higa, Luiza Mendonça

    Memorias do Instituto Oswaldo Cruz

    2022  Volume 116, Page(s) e210176

    Abstract: Background: During routine Coronavirus disease 2019 (COVID-19) diagnosis, an unusually high viral load was detected by reverse transcription real-time polymerase chain reaction (RT-qPCR) in a nasopharyngeal swab sample collected from a patient with ... ...

    Abstract Background: During routine Coronavirus disease 2019 (COVID-19) diagnosis, an unusually high viral load was detected by reverse transcription real-time polymerase chain reaction (RT-qPCR) in a nasopharyngeal swab sample collected from a patient with respiratory and neurological symptoms who rapidly succumbed to the disease. Therefore we sought to characterise the infection.
    Objectives: We aimed to determine and characterise the etiological agent responsible for the poor outcome.
    Methods: Classical virological methods, such as plaque assay and plaque reduction neutralisation test combined with amplicon-based sequencing, as well as a viral metagenomic approach, were performed to characterise the etiological agents of the infection.
    Findings: Plaque assay revealed two distinct plaque phenotypes, suggesting either the presence of two severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) strains or a productive coinfection of two different species of virus. Amplicon-based sequencing did not support the presence of any SARS-CoV-2 genetic variants that would explain the high viral load and suggested the presence of a single SARS-CoV-2 strain. Nonetheless, the viral metagenomic analysis revealed that Coronaviridae and Herpesviridae were the predominant virus families within the sample. This finding was confirmed by a plaque reduction neutralisation test and PCR.
    Main conclusions: We characterised a productive coinfection of SARS-CoV-2 and Herpes simplex virus 1 (HSV-1) in a patient with severe symptoms that succumbed to the disease. Although we cannot establish the causal relationship between the coinfection and the severity of the clinical case, this work serves as a warning for future studies focused on the interplay between SARS-CoV-2 and HSV-1 coinfection and COVID-19 severity.
    MeSH term(s) COVID-19 ; Coinfection ; Herpesvirus 1, Human/genetics ; Humans ; Real-Time Polymerase Chain Reaction ; SARS-CoV-2
    Language English
    Publishing date 2022-01-10
    Publishing country Brazil
    Document type Journal Article
    ZDB-ID 953293-6
    ISSN 1678-8060 ; 0074-0276
    ISSN (online) 1678-8060
    ISSN 0074-0276
    DOI 10.1590/0074-02760210176
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: Yellow fever virus is susceptible to sofosbuvir both in vitro and in vivo.

    de Freitas, Caroline S / Higa, Luiza M / Sacramento, Carolina Q / Ferreira, André C / Reis, Patrícia A / Delvecchio, Rodrigo / Monteiro, Fabio L / Barbosa-Lima, Giselle / James Westgarth, Harrison / Vieira, Yasmine Rangel / Mattos, Mayara / Rocha, Natasha / Hoelz, Lucas Villas Bôas / Leme, Rennan Papaleo Paes / Bastos, Mônica M / Rodrigues, Gisele Olinto L / Lopes, Carla Elizabeth M / Queiroz-Junior, Celso Martins / Lima, Cristiano X /
    Costa, Vivian V / Teixeira, Mauro M / Bozza, Fernando A / Bozza, Patrícia T / Boechat, Nubia / Tanuri, Amilcar / Souza, Thiago Moreno L

    PLoS neglected tropical diseases

    2019  Volume 13, Issue 1, Page(s) e0007072

    Abstract: Yellow fever virus (YFV) is a member of the Flaviviridae family. In Brazil, yellow fever (YF) cases have increased dramatically in sylvatic areas neighboring urban zones in the last few years. Because of the high lethality rates associated with infection ...

    Abstract Yellow fever virus (YFV) is a member of the Flaviviridae family. In Brazil, yellow fever (YF) cases have increased dramatically in sylvatic areas neighboring urban zones in the last few years. Because of the high lethality rates associated with infection and absence of any antiviral treatments, it is essential to identify therapeutic options to respond to YFV outbreaks. Repurposing of clinically approved drugs represents the fastest alternative to discover antivirals for public health emergencies. Other Flaviviruses, such as Zika (ZIKV) and dengue (DENV) viruses, are susceptible to sofosbuvir, a clinically approved drug against hepatitis C virus (HCV). Our data showed that sofosbuvir docks onto YFV RNA polymerase using conserved amino acid residues for nucleotide binding. This drug inhibited the replication of both vaccine and wild-type strains of YFV on human hepatoma cells, with EC50 values around 5 μM. Sofosbuvir protected YFV-infected neonatal Swiss mice and adult type I interferon receptor knockout mice (A129-/-) from mortality and weight loss. Because of its safety profile in humans and significant antiviral effects in vitro and in mice, Sofosbuvir may represent a novel therapeutic option for the treatment of YF. Key-words: Yellow fever virus; Yellow fever, antiviral; sofosbuvir.
    MeSH term(s) Animals ; Antiviral Agents/pharmacology ; Cercopithecus aethiops ; Disease Models, Animal ; Drug Resistance, Viral ; Hep G2 Cells ; Humans ; Mice ; Mice, Knockout ; RNA, Viral/blood ; RNA, Viral/drug effects ; RNA, Viral/genetics ; Sofosbuvir/pharmacology ; Vero Cells ; Yellow Fever/blood ; Yellow Fever/drug therapy ; Yellow Fever/pathology ; Yellow Fever/virology ; Yellow fever virus/drug effects ; Yellow fever virus/genetics
    Chemical Substances Antiviral Agents ; RNA, Viral ; Sofosbuvir (WJ6CA3ZU8B)
    Language English
    Publishing date 2019-01-30
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2429704-5
    ISSN 1935-2735 ; 1935-2727
    ISSN (online) 1935-2735
    ISSN 1935-2727
    DOI 10.1371/journal.pntd.0007072
    Database MEDical Literature Analysis and Retrieval System OnLINE

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