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1.
Virology ; 596: 110095, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38761641

ABSTRACT

Dengue virus (DENV) is a major global health concern, causing millions of infections annually. Understanding the cellular response to DENV infection is crucial for developing effective therapies. This study provides an in-depth analysis of the cellular response to Dengue virus (DENV) infection, with a specific focus on the interplay between microRNAs (miRNAs), apoptosis, and viral load across different DENV serotypes. Utilizing a variety of cell lines infected with four DENV serotypes, the research methodically quantifies viral load, and the expression levels of miRNA-15, miRNA-16, and BCL2 protein, alongside measuring apoptosis markers. Methodologically, the study employs quantitative PCR for viral load and miRNA expression analysis, and Western blot for apoptosis and BCL2 detection, with a statistical framework that includes ANOVA and correlation analysis to discern significant differences and relationships. The findings reveal that despite similar viral loads across DENV serotypes, DENV-2 exhibits a marginally higher load. A notable upregulation of miRNA-15 and miRNA-16 correlates positively with increased viral load, suggesting their potential role in modulating viral replication. Concurrently, a marked activation of caspases 3 and 7, along with changes in BCL2 protein levels, underscores the role of apoptosis in the cellular response to DENV infection. Conclusively, the study enhances the understanding of miRNA involvement in DENV pathogenesis, highlighting miRNA-15 and miRNA-16 as potential regulatory agents in viral replication and apoptosis. These findings pave the way for further exploration into miRNA-based therapeutic strategies against DENV infection.


Subject(s)
Apoptosis , Dengue Virus , Dengue , MicroRNAs , Proto-Oncogene Proteins c-bcl-2 , Viral Load , Virus Replication , MicroRNAs/genetics , MicroRNAs/metabolism , Dengue Virus/physiology , Dengue Virus/genetics , Humans , Proto-Oncogene Proteins c-bcl-2/genetics , Proto-Oncogene Proteins c-bcl-2/metabolism , Dengue/virology , Cell Line , Caspase 3/metabolism , Caspase 3/genetics , Caspase 7/metabolism , Caspase 7/genetics , Serogroup
2.
PLoS One ; 19(4): e0299993, 2024.
Article in English | MEDLINE | ID: mdl-38568963

ABSTRACT

The selection of proper reference genes is critical for accurate gene expression analysis in all fields of biological and medical research, mainly because there are many distinctions between different tissues and specimens. Given this variability, even in known classic reference genes, demands of a comprehensive analysis platform is needed to identify the most suitable genes for each study. For this purpose, we present an analysis tool for assisting in decision-making in the analysis of reverse transcription-quantitative polymerase chain reaction (RT-qPCR) data. EndoGeneAnalyzer, an open-source web tool for reference gene analysis in RT-qPCR studies, was used to compare the groups/conditions under investigation. This interactive application offers an easy-to-use interface that allows efficient exploration of datasets. Through statistical and stability analyses, EndoGeneAnalyzer assists in the select of the most appropriate reference gene or set of genes for each condition. It also allows researchers to identify and remove unwanted outliers. Moreover, EndoGeneAnalyzer provides a graphical interface to compare the evaluated groups, providing a visually informative differential analysis.


Subject(s)
Gene Expression Profiling , Real-Time Polymerase Chain Reaction , Reference Standards , Reverse Transcriptase Polymerase Chain Reaction
3.
Genes (Basel) ; 14(10)2023 10 05.
Article in English | MEDLINE | ID: mdl-37895256

ABSTRACT

COVID-19 is an infectious disease caused by coronavirus 2 of the severe acute syndrome (SARS-CoV-2). Single nucleotide polymorphisms (SNPs) in genes, such as TLR2, responsible for an effective human immune response, can change the course of infection. The objective of this article was to verify associations between epidemiological factors and TLR2 SNP rs3804100 (Thymine [T] > Cytosine [C]) in professionals from Health Institutions (HI) who worked during the first pandemic wave and COVID-19. A case-control study was conducted with Belém-PA HI workers (Northern Brazil), divided into symptomatology groups (Asymptomatic-AS; n = 91; and Symptomatic-SI; n = 123); and severity groups classified by Chest Computerized Tomography data (symptomatic with pulmonary involvement-SCP; n = 35; symptomatic without pulmonary involvement-SSP; n = 8). Genotyping was performed by Sanger sequencing, and Statistical Analysis was conducted through the SPSS program. Bioinformatics servers predicted the biological functions of the TLR2 SNP. There were associations between the presence of comorbidities and poor prognosis of COVID-19 (especially between symptomatology and severity of COVID-19 and overweight and obesity) and between the sickness in family members and kinship (related to blood relatives). The homozygous recessive (C/C) genotype was not found, and the frequency of the mutant allele (C) was less than 10% in the cohort. No significant associations were found for this SNP in this cohort. The presence of SNP was indicated to be benign and causes a decrease in the stability of the TLR2 protein. These data can help the scientific community and medicine find new forms of COVID-19 containment.


Subject(s)
COVID-19 , Polymorphism, Single Nucleotide , Humans , Toll-Like Receptor 2/genetics , Genetic Predisposition to Disease , Case-Control Studies , Pandemics , Brazil/epidemiology , COVID-19/epidemiology , COVID-19/genetics , SARS-CoV-2/genetics
4.
Pharmaceuticals (Basel) ; 16(9)2023 Aug 30.
Article in English | MEDLINE | ID: mdl-37765037

ABSTRACT

Natural compounds with pharmacological activity, flavonoids have been the subject of an exponential increase in studies in the field of scientific research focused on therapeutic purposes due to their bioactive properties, such as antioxidant, anti-inflammatory, anti-aging, antibacterial, antiviral, neuroprotective, radioprotective, and antitumor activities. The biological potential of flavonoids, added to their bioavailability, cost-effectiveness, and minimal side effects, direct them as promising cytotoxic anticancer compounds in the optimization of therapies and the search for new drugs in the treatment of cancer, since some extensively antineoplastic therapeutic approaches have become less effective due to tumor resistance to drugs commonly used in chemotherapy. In this review, we emphasize the antitumor properties of tangeretin, a flavonoid found in citrus fruits that has shown activity against some hallmarks of cancer in several types of cancerous cell lines, such as antiproliferative, apoptotic, anti-inflammatory, anti-metastatic, anti-angiogenic, antioxidant, regulatory expression of tumor-suppressor genes, and epigenetic modulation.

5.
Curr Issues Mol Biol ; 45(6): 4589-4599, 2023 May 26.
Article in English | MEDLINE | ID: mdl-37367040

ABSTRACT

The World Health Organization has estimated the annual occurrence of approximately 392 million dengue virus (DENV) infections in more than 100 countries where the virus is endemic, which represents a serious threat to humanity. DENV is a serologic group with four distinct serotypes (DENV-1, DENV-2, DENV-3, and DENV-4) belonging to the genus Flavivirus, in the family Flaviviridae. Dengue is the most widespread mosquito-borne disease in the world. The ~10.7 kb DENV genome encodes three structural proteins (capsid (C), pre-membrane (prM), and envelope (E)) and seven non-structural (NS) proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B, and NS5). The NS1 protein is a membrane-associated dimer and a secreted, lipid-associated hexamer. Dimeric NS1 is found on membranes both in cellular compartments and cell surfaces. Secreted NS1 (sNS1) is often present in patient serum at very high levels, which correlates with severe dengue symptoms. This study was conducted to discover how the NS1 protein, microRNAs-15/16 (miRNAs-15/16), and apoptosis are related during DENV-4 infection in human liver cell lines. Huh 7.5 and HepG2 cells were infected with DENV-4, and miRNAs-15/16, viral load, NS1 protein, and caspases-3/7 were quantified after different durations of infection. This study demonstrated that miRNAs-15/16 were overexpressed during the infection of HepG2 and Huh 7.5 cells with DENV-4 and had a relationship with NS1 protein expression, viral load, and the activity of caspases-3/7, thus making these miRNAs potential injury markers during DENV infection in human hepatocytes.

6.
Einstein (Sao Paulo) ; 21: eAO0160, 2023.
Article in English | MEDLINE | ID: mdl-37255058

ABSTRACT

OBJECTIVE: This study verified the replication efficiency of the Rocio virus in a primary culture of mouse neural cells. METHODS: Mixed primary cultures (neurons/glia) obtained from the brains of newborn isogenic BALB/c mice were inoculated with Rocio virus on the 7 th day of culture, and the development of cytopathogenic effects was monitored. The infection was confirmed via immunocytochemistry (anti-ROCV), while viral replication was quantified in infected primary cultures. The titration method used depended on the infection period. RESULTS: Rocio virus efficiently infected primary cultured neural cells, with the highest viral titer causing cytopathic changes was observed at 2 days post infection. The virus-infected primary culture survived for up to 7 days post infection, and viral load quantitation showed viral replication kinetics compatible with the cell death kinetics of cultures. CONCLUSION: The findings of this study suggest that mouse neural cell primary cultures support Rocio virus replication and could be used as an alternative system for studying Flavivirus infection in the central nervous system.


Subject(s)
Flavivirus Infections , Flavivirus , Animals , Mice , Flavivirus Infections/metabolism , Flavivirus Infections/pathology , Brain , Neurons/metabolism , Neurons/pathology , Cells, Cultured
7.
Einstein (Säo Paulo) ; 21: eAO0160, 2023. graf
Article in English | LILACS-Express | LILACS | ID: biblio-1440063

ABSTRACT

ABSTRACT Objective This study verified the replication efficiency of the Rocio virus in a primary culture of mouse neural cells. Methods Mixed primary cultures (neurons/glia) obtained from the brains of newborn isogenic BALB/c mice were inoculated with Rocio virus on the 7 th day of culture, and the development of cytopathogenic effects was monitored. The infection was confirmed via immunocytochemistry (anti-ROCV), while viral replication was quantified in infected primary cultures. The titration method used depended on the infection period. Results Rocio virus efficiently infected primary cultured neural cells, with the highest viral titer causing cytopathic changes was observed at 2 days post infection. The virus-infected primary culture survived for up to 7 days post infection, and viral load quantitation showed viral replication kinetics compatible with the cell death kinetics of cultures. Conclusion The findings of this study suggest that mouse neural cell primary cultures support Rocio virus replication and could be used as an alternative system for studying Flavivirus infection in the central nervous system.

8.
Front Cell Infect Microbiol ; 13: 1320701, 2023.
Article in English | MEDLINE | ID: mdl-38173795

ABSTRACT

The objective of this article was to verify associations between the SNPs rs3775291 (Cytosine [C]>Thymine [T]) and rs3775290 (C>T) of TLR3 in professionals from Health Institutions (HI) who worked during the first pandemic wave and COVID-19. A case-control study was carried out with workers from HI in Belém-PA, Brazil, divided into symptomatology groups (Asymptomatic-AS, n=91; and Symptomatic-SI, n=121), and severity groups, classified by Chest CT scan (symptomatic with lung involvement - SCP, n=34; symptomatic without lung involvement - SSP, n=8). Genotyping was performed by Sanger sequencing and statistical analysis was performed using the SPSS program. In the analysis of SNP rs3775291, the homozygous recessive genotype (T/T) was not found and the frequency of the mutant allele (T) was less than 2% in the cohort. For the rs3775290 SNP, the frequency of the mutant allele (T) was greater than 42% in the cohort. No significant associations were found for these SNPs in this cohort (N= 212 individuals). The scientific community and physicians can use these facts to find new methods of managing COVID-19.


Subject(s)
COVID-19 , Polymorphism, Single Nucleotide , Humans , Toll-Like Receptor 3/genetics , Genetic Predisposition to Disease , Case-Control Studies , Brazil/epidemiology , COVID-19/genetics , Genotype
9.
Genes (Basel) ; 13(11)2022 11 18.
Article in English | MEDLINE | ID: mdl-36421821

ABSTRACT

The COVID-19 pandemic initiated a race to determine the best measures to control the disease and to save as many people as possible. Efforts to implement social distancing, the use of masks, and massive vaccination programs turned out to be essential in reducing the devastating effects of the pandemic. Nevertheless, the high mutation rates of SARS-CoV-2 challenge the vaccination strategy and maintain the threat of new outbreaks due to the risk of infection surges and even lethal variations able to resist the effects of vaccines and upset the balance. Most of the new therapies tested against SARS-CoV-2 came from already available formulations developed to treat other diseases, so they were not specifically developed for SARS-CoV-2. In parallel, the knowledge produced regarding the molecular mechanisms involved in this disease was vast due to massive efforts worldwide. Taking advantage of such a vast molecular understanding of virus genomes and disease mechanisms, a targeted molecular therapy based on siRNA specifically developed to reach exclusive SARS-CoV-2 genomic sequences was tested in a non-transformed human cell model. Since coronavirus can escape from siRNA by producing siRNA inhibitors, a complex strategy to simultaneously strike both the viral infectious mechanism and the capability of evading siRNA therapy was developed. The combined administration of the chosen produced siRNA proved to be highly effective in successfully reducing viral load and keeping virus replication under control, even after many days of treatment, unlike the combinations of siRNAs lacking this anti-anti-siRNA capability. Additionally, the developed therapy did not harm the normal cells, which was demonstrated because, instead of testing the siRNA in nonhuman cells or in transformed human cells, a non-transformed human thyroid cell was specifically chosen for the experiment. The proposed siRNA combination could reduce the viral load and allow the cellular recovery, presenting a potential innovation for consideration as an additional strategy to counter or cope COVID-19.


Subject(s)
COVID-19 , SARS-CoV-2 , Humans , SARS-CoV-2/genetics , Pandemics , Virus Replication/genetics , Genome, Viral , RNA, Small Interfering/genetics
10.
Preprint in Portuguese | SciELO Preprints | ID: pps-4314

ABSTRACT

The present study aimed to develop a children's book for learning about viral immunization in elementary school. Thus, the object of alternative teaching construct through bibliographic research, copy the content, book production and illustrations, dissemination, and collection of readers' satisfaction. The development of playful-pedagogical methodologies, such as the book "Welcome to the Microscopic Universe," is an alternative to alleviate students' lack of playful resources. In return, they have the opportunity to assimilate pleasantly of body and defense cells against viruses. In this regard, emphasized that children's literature is an open universe of possibilities and content. Each day, new authors create increasingly effective and technological teaching methodologies, transforming learning through books into an interaction of exchange of information between the subject and the environment in which they live.


El presente estudio tuvo como objetivo desarrollar la elaboración de un libro infantil para el aprendizaje de la inmunización viral en la escuela primaria. Así, el objeto de la enseñanza alternativa se construyó a través de la investigación bibliográfica, los contenidos de copia, la producción e ilustraciones de libros, la difusión y recogida de satisfacción de los lectores. El desarrollo de metodologías lúdico-pedagógicas, como el libro "Bienvenidos al Universo Microscópico" es una alternativa para paliar la falta de recursos lúdicos para los alumnos, y a cambio, tienen la oportunidad de asimilar de manera amena, de su cuerpo y sus células de defensa contra los virus. En este sentido, se destaca que la literatura infantil es un universo abierto de posibilidades y contenidos, que cada día, nuevos autores crean metodologías de enseñanza cada vez más efectivas y tecnológicas, transformando el aprendizaje a través de los libros en una interacción de intercambio de información entre el sujeto y el entorno en el que viven.


O presente trabalho objetivou o desenvolvimento de um livro infantil para a aprendizagem sobre a imunização viral no ensino fundamental.  Sendo assim, o objeto de ensino alternativo foi construído através de pesquisa bibliográfica, conteúdo do exemplar, produção do livro e ilustrações, divulgação e coleta de satisfação dos leitores. O desenvolvimento de metodologias lúdico-pedagógicas, como o livro " Bem-Vindos ao Universo Microscópico" é uma alternativa para amenizar a falta de recursos lúdicos para os alunos, e, em contrapartida, os mesmos terem a oportunidade de assimilar de forma prazerosa, a respeito do seu corpo e de suas células de defesa contra os vírus. Neste intuito, ressalta-se que a literatura infantil é um universo aberto de possibilidades e conteúdos, que a cada dia, novos autores criam metodologias de ensino cada vez mais eficaz e tecnológicas, transformando a aprendizagem através dos livros uma interação de troca de informações entre o sujeito e o meio em que se vive.

11.
Acta Trop ; 227: 106285, 2022 Mar.
Article in English | MEDLINE | ID: mdl-34921765

ABSTRACT

Cecropins and defensins are the main classes of antimicrobial peptides in the mosquito innate immune system, acting against bacteria, fungi and protozoa. There is a knowledge gap concerning these peptide genes in anopheline mosquitoes from the Brazilian Amazon. Thus, this work aimed to describe molecular techniques for detecting the genes encoding the antimicrobial peptides cecropin A (CecA) and defensin in Anopheles darlingi mosquitoes and to perform molecular phylogeny of the sequenced genes using the maximum likelihood method and Bayesian inference with other species from different geographic areas. Our results show, for the first time, a molecular biology method for detecting CecA and defensin in Anopheles darlingi that allows for the use of these molecular markers for phylogenetic analysis in anopheline species, separating the species into single and monophyletic clades.


Subject(s)
Anopheles , Cecropins , Animals , Anopheles/genetics , Antimicrobial Peptides , Bayes Theorem , Cecropins/genetics , Cecropins/pharmacology , Defensins/genetics , Defensins/pharmacology , Phylogeny
12.
Microorganisms ; 11(1)2022 Dec 21.
Article in English | MEDLINE | ID: mdl-36677318

ABSTRACT

Visceral leishmaniasis (VL), also known as kala-azar, is an anthropozoonotic disease affecting human populations on five continents. Aetiologic agents belong to the Leishmania (L.) donovani complex. Until the 1990s, three leishmanine parasites comprised this complex: L. (L.) donovani Laveran & Mesnil 1903, L. (L.) infantum Nicolle 1908, and L. (L.) chagasi Lainson & Shaw 1987 (=L. chagasi Cunha & Chagas 1937). The VL causal agent in the New World (NW) was previously identified as L. (L.) chagasi. After the development of molecular characterization, however, comparisons between L. (L.) chagasi and L. (L.) infantum showed high similarity, and L. (L.) chagasi was then regarded as synonymous with L. (L.) infantum. It was, therefore, suggested that L. (L.) chagasi was not native to the NW but had been introduced from the Old World by Iberian colonizers. However, in light of ecological evidence from the NW parasite's enzootic cycle involving a wild phlebotomine vector (Lutzomyia longipalpis) and a wild mammal reservoir (the fox, Cerdocyon thous), we have recently analyzed by molecular clock comparisons of the DNA polymerase alpha subunit gene the whole-genome sequence of L. (L.) infantum chagasi of the most prevalent clinical form, atypical dermal leishmaniasis (ADL), from Honduras (Central America) with that of the same parasite from Brazil (South America), as well as those of L. (L.) donovani (India) and L. (L.) infantum (Europe), which revealed that the Honduran parasite is older ancestry (382,800 ya) than the parasite from Brazil (143,300 ya), L. (L.) donovani (33,776 ya), or L. (L.) infantum (13,000 ya). In the present work, we have now amplified the genomic comparisons among these leishmanine parasites, exploring mainly the variations in the genome for each chromosome, and the number of genomic SNPs for each chromosome. Although the results of this new analysis have confirmed a high genomic similarity (~99%) among these parasites [except L. (L.) donovani], the Honduran parasite revealed a single structural variation on chromosome 17, and the highest frequency of genomic SNPs (more than twice the number seen in the Brazilian one), which together to its extraordinary ancestry (382,800 ya) represent strong evidence that L. (L.) chagasi/L. (L.) infantum chagasi is, in fact, native to the NW, and therefore with valid taxonomic status. Furthermore, the Honduran parasite, the most ancestral viscerotropic leishmanine parasite, showed genomic and clinical taxonomic characteristics compatible with a new Leishmania species causing ADL in Central America.

13.
Parasitol Int ; 81: 102273, 2021 Apr.
Article in English | MEDLINE | ID: mdl-33333246

ABSTRACT

The clinical-immunological spectrum of human Leishmania (L.) infantum chagasi-infections in the Brazilian Amazon has been defined using DTH/IFAT-IgG immune assays and the clinical statuses of infected individuals, revealing five profiles: three asymptomatic [Asymptomatic Infection (AI), Subclinical Resistant Infection (SRI), and Indeterminate Initial Infection (III)], and two symptomatic profiles [Subclinical Oligosymptomatic Infection (SOI) and Symptomatic Infection (SI = American visceral leishmaniasis/AVL)]. We evaluated the diagnostic potential of urine qPCR over the entire spectrum of infection. Resine Instagene Matrix® was used for DNA extraction from urinary sediment, with amplification carried out using SYBR® Green Taq with the RV1 and RV2 primers. We examined urine samples from 151 individuals from an endemic area of AVL in Pará State in the Brazilian Amazon, including: 91 (60.3%) with diagnoses of previous infections [13 (14.3%) sharing the AI profile, 13 (14.3%) with the SRI profile, 43 (47.2%) with III, 12 (13.2%) with SI (treated AVL), and 10 (11%) with SI (untreated AVL)]; sixty (39.7%) were DTH(-)/IFAT-IgG(-) (the uninfected group). The urine qPCR was positive in 61.5% of both the AI and SRI profiles, 65% of the III profile, 50% of treated AVL, 100% of untreated AVL, and 6.7% of the uninfected group. Those results confirmed the urine qPCR diagnosis in 100% of untreated AVL cases as well as in more than 60% of the cases with asymptomatic AI, SRI, and III profiles - indicating it as a promising tool for monitoring the evolution of human L. (L.) infantum chagasi-infections in endemic areas.


Subject(s)
Leishmania infantum/isolation & purification , Leishmaniasis, Visceral/diagnosis , Adult , Aged , Aged, 80 and over , Asymptomatic Infections , Brazil , Female , Humans , Leishmaniasis, Visceral/parasitology , Leishmaniasis, Visceral/urine , Male , Middle Aged , Real-Time Polymerase Chain Reaction , Young Adult
14.
Viruses ; 11(9)2019 08 22.
Article in English | MEDLINE | ID: mdl-31443500

ABSTRACT

Because of its ecological characteristics, the Caxiuanã National Forest (FLONA) is a potential area of arbovirus circulation. The present study aimed to investigate the occurrence of arbovirus transmission cycles at FLONA de Caxiuanã. Five field trips were performed to capture mosquitoes and sylvatic vertebrates. For these vertebrates, we attempted viral isolation by cell monolayer inoculation from blood, and hemagglutination inhibition and further seroneutralization assays from sera. For mosquitoes, we performed tests of viral genome detection. A total of 338 vertebrates were captured, and the greatest representative was birds (251/74.26%). A total of 16,725 mosquitoes were captured, distributed among 56 species. There were no viruses isolated by newborn mouse inoculation. Among birds, antibodies against Ilheus virus were the most prevalent. Catu virus, Caraparu virus, and Mucambo virus were the most prevalent among mammals and reptiles. Fragments of Mucambo virus, Ilheus virus, Bussuquara virus, and Rocio virus genome were detected in a pool of mosquito samples. These results of the study suggest the occurrence of arbovirus transmission cycles in the FLONA of Caxiuanã. The proximity of human populations with elements, involved in transmission cycles, makes surveillance necessary in this population to avoid dispersion of arboviruses to naïve locations.


Subject(s)
Arbovirus Infections/transmission , Arboviruses , Birds/virology , Mammals/virology , Mosquito Vectors/virology , Animals , Arboviruses/genetics , Arboviruses/isolation & purification , Biodiversity , Brazil , Culicidae/virology , Genome, Viral , Hemagglutination Inhibition Tests , Rainforest
15.
Article in English | MEDLINE | ID: mdl-30863824

ABSTRACT

We identified a strain of Alphacoronavirus 1, FCoV-SB22, from a pool of fecal samples from domestic cats from a rural settlement in the municipality of Santa Bárbara, Pará, Brazil. The nucleotide identity with feline coronavirus was 91.5%. The present study reports the first complete genome sequence of a feline coronavirus from Brazil.

16.
J Histochem Cytochem ; 67(6): 419-439, 2019 06.
Article in English | MEDLINE | ID: mdl-30924711

ABSTRACT

Peripheral inflammatory stimuli increase proinflammatory cytokines in the bloodstream and central nervous system and activate microglial cells. Here we tested the hypothesis that contrasting environments mimicking sedentary and active lives would be associated with differential microglial morphological responses, inflammatory cytokines concentration, and virus load in the peripheral blood. For this, mice were maintained either in standard (standard environment) or enriched cages (enriched environment) and then subjected to a single (DENV1) serotype infection. Blood samples from infected animals showed higher viral loads and higher tumor necrosis factor-α (TNFα) mRNA concentrations than control subjects. Using an unbiased stereological sampling approach, we selected 544 microglia from lateral septum for microscopic 3D reconstruction. Morphological complexity contributed most to cluster formation. Infected groups exhibited significant increase in the microglia morphological complexity and number, despite the absence of dengue virus antigens in the brain. Two microglial phenotypes (type I with lower and type II with higher morphological complexity) were found in both infected and control groups. However, microglia from infected mice maintained in enriched environment showed only one morphological phenotype. Two-way ANOVA revealed that environmental changes and infection influenced type-I and II microglial morphologies and number. Environmental enrichment and infection interactions may contribute to microglial morphological change to a point that type-I and II morphological phenotypes could no longer be distinguished in infected mice from enriched environment. Significant linear correlation was found between morphological complexity and TNFα peripheral blood. Our findings demonstrated that sedentary-like and active murine models exhibited differential microglial responses and peripheral inflammation to systemic non-neurotropic infections with DENV1 virus.


Subject(s)
Dengue Virus/physiology , Dengue/metabolism , Dengue/pathology , Microglia/pathology , Tumor Necrosis Factor-alpha/metabolism , Viral Load , Animals , Disease Models, Animal , Female , Gene Expression Regulation , Humans , Mice
17.
Arch Virol ; 164(4): 1187-1192, 2019 Apr.
Article in English | MEDLINE | ID: mdl-30725182

ABSTRACT

Yellow fever virus (YFV) penetrates the skin through the bite of a vector mosquito and spreads to various organs, mainly the liver, where it causes lesions and induces necrosis and apoptosis. We evaluated the mRNA expression of various cytokines and the activation of caspases in HepG2 cells infected with YFV. We observed that interferon-α (IFN-α) expression decreased and IFN-ß, transforming growth factor (TGF)-ß IIIR, interleukin (IL)-6, and IL-8 expression increased in cells infected with genotype 1. In contrast, TNF-α expression increased in cells infected with genotype 2 but not with genotype 1. This provides insights into the role of cytokine regulation in yellow fever.


Subject(s)
Caspase 3/metabolism , Caspase 7/metabolism , Cytokines/genetics , Liver Neoplasms/genetics , Yellow fever virus/physiology , Caspase 3/genetics , Caspase 7/genetics , Cell Line , Cytokines/metabolism , Hep G2 Cells , Host-Pathogen Interactions , Humans , Interferon-gamma/genetics , Interferon-gamma/metabolism , Liver Neoplasms/metabolism , Liver Neoplasms/virology , Transforming Growth Factor beta/genetics , Transforming Growth Factor beta/metabolism , Yellow fever virus/genetics
18.
Article in English | MEDLINE | ID: mdl-30637382

ABSTRACT

A proposed new strain of canine Kobuvirus was identified in fecal samples of domestic dogs from a rural community located in the municipality of Peixe-Boi, Pará, Brazil. The nucleotide identity was 92.3% similar to other representatives of the family Picornaviridae, genus Kobuvirus, and species Aichivirus A, which suggests that this is possibly a new strain within this species.

19.
Genomics ; 111(4): 607-611, 2019 07.
Article in English | MEDLINE | ID: mdl-29581026

ABSTRACT

Mosquitoes as Sabethes chloropterus, Sabethes glaucodaemon, Sabethes belisarioi are species of medical and epidemiological importance for arboviruses transmission such as yellow fever and St. Louis encephalitis. Despite this, no information about these three species mitochondrial DNA has been found in literature. Our study presents a mitochondrial genome description, including identity, SNPs, mutation rate, and phylogeny analysis using COX1, COX2, NADH4, NADH5, CYOB genes. The Sa. chloropterus, Sa. glaucodaemon and Sa. belisaroi mitochondrial genome sizes 15.609 bp, 15.620 bp, 15.907 bp, respectively, with 37 functional genes, presenting about 4.982 single nucleotide polymorphisms and 13.291 identical sites between them, besides all genes with dN/dS < 1 ratio, and also a greater approximation between Sa. glaucodaemon and Sa. chloropterus than with Sa. belisarioi. Due to the importance of mitochondrial DNA for population structure studies, evolution, and others, we expect that this data can contribute to other studies related to these mosquitoes and their viruses.


Subject(s)
Culicidae/genetics , Genome, Mitochondrial , Phylogeny , Animals , Culicidae/classification , Electron Transport Complex I/genetics , Electron Transport Complex IV/genetics , Insect Proteins/genetics , Polymorphism, Genetic
20.
Viral Immunol ; 31(8): 583-588, 2018 10.
Article in English | MEDLINE | ID: mdl-29878881

ABSTRACT

Zika virus (ZIKV) is an arbovirus belonging to the genus Flavivirus (Flaviviridae). ZIKV infection is associated with alterations in various organs, including the liver, lungs, and kidneys. Studies on the influence of posttranscriptional control on viral infections have demonstrated that microRNAs (miRNAs) interfere with different stages of the replicative cycle of several viruses and may influence the disease outcome. To shed light on ZIKV-induced regulation of host miRNA-processing machinery in the above organs, we analyzed the expression of genes encoding key proteins of the miRNA pathway in different ZIKV-infected continuous primate cell lineages (HepG2, A549, and MA104) by reverse-transcription quantitative polymerase chain reaction (RT-qPCR). Expression of the genes encoding the miRNA-related proteins DGCR8, Ago1, and Ago3 in HepG2 cells and Drosha, Dicer, Ago2, and Ago3 in A549 and MA104 cells was significantly altered in the presence of ZIKV. Our results suggest that ZIKV modulates miRNA levels during infection in liver, lung, and kidney cells, which may be an additional mechanism of host cell subversion in these organs.


Subject(s)
Kidney/cytology , Liver/cytology , Lung/cytology , MicroRNAs/genetics , Zika Virus/immunology , Animals , Cell Lineage , Chlorocebus aethiops , Gene Expression Regulation , Hep G2 Cells , Host-Pathogen Interactions/genetics , Humans , Kidney/virology , Liver/virology , Lung/virology , Virus Replication
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