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1.
Emerg Infect Dis ; 25(8): 1477-1484, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31310224

RESUMO

We characterized natural vertical transmission of Zika virus in pools of Aedes aegypti larvae hatched from eggs collected in Jojutla, Morelos, Mexico. Of the 151 pools analyzed, 17 tested positive for Zika virus RNA; infectious Zika virus was successfully isolated from 1 of the larvae pools (31N) in C6/36 cells. Real-time quantitative PCR and indirect immunofluorescence assays confirmed the identity of the isolate, named Zika virus isolate 31N; plaque assays in Vero cells demonstrated the isolate's infectivity in a mammalian cell line. We obtained the complete genome of Zika virus isolate 31N by next-generation sequencing and identified 3 single-nucleotide variants specific to Zika virus isolate 31N using the meta-CATS tool. These results demonstrate the occurrence of natural vertical transmission of Zika virus in wild Ae. aegypti mosquitoes and suggest that this transmission mode could aid in the spread and maintenance of Zika virus in nature.


Assuntos
Aedes/virologia , Mosquitos Vetores/virologia , Zika virus/fisiologia , Animais , Linhagem Celular , Células Cultivadas , Chlorocebus aethiops , Microbiologia Ambiental , Genoma Viral , Humanos , Transmissão Vertical de Doenças Infecciosas , Larva , México/epidemiologia , Filogenia , Vigilância em Saúde Pública , Células Vero , Carga Viral , Ensaio de Placa Viral , Sequenciamento Completo do Genoma , Zika virus/classificação , Infecção por Zika virus/transmissão , Infecção por Zika virus/virologia
2.
Eur J Clin Invest ; 49(5): e13085, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30740673

RESUMO

BACKGROUND: Atherosclerosis represents a cardiovascular risk. Chronic inflammation is a key factor for atherogenic progression. Neutrophil-to-lymphocyte ratio (NLR) has been proposed as a novel biomarker for cardiovascular risks. We aimed to explore whether NLR was related to surrogate pro-atherogenic promoters driving atherogenic progression, as measured by carotid intima-media thickness (CIMT). STUDY DESIGN: Thirty-one patients with obesity candidates for bariatric surgery were recruited from Centro Médico Nacional "20 de Noviembre", ISSSTE, Mexico City. The results are part of the "CROP" study (NCT03561987). NLR was calculated from routine complete blood count, and its relation with plasma pro-inflammatory mediators (hsCRP, TNF-α and IL-1ß), adipokines (adiponectin and leptin), adiposity markers (visceral adipose tissue [VAT] determined from CT scan image and VAT individual adipocyte area at histological sample) and CIMT were determined. RESULTS: Neutrophil-to-lymphocyte ratio correlated with hsCRP (Spearman's r = 0.70 [95% CI 0.46 to 0.85], P < 0.01), TNF-α (r = 0.69 [0.44 to 0.84], P < 0.0001) and adiponectin (r = -0.69 [-0.84 to -0.45], P < 0.03), as well as with VAT individual adipocyte area (r = 0.64 [0.37 to 0.81], P < 0.0001) and with VAT area (r = 0.43; [0.07 to 0.68], P < 0.01). Leptin and adiponectin showed further independent association with higher NLR (multivariate regression analysis OR 7.9 [95% CI 1.1 to 56.2] P = 0.03 and 0.1 [0.01 to 1.0] P = 0.05, respectively). Moreover, NLR distribution significantly varied between subgroups divided according to progressive CIMT (P = 0.05); whereas adiponectin and VAT adipocyte area associated with CIMT > 0.9 mm (univariate analysis OR 0.1 [0.01 to 1.0] P = 0.05 and 13.1 [1.4 to 126.3] P = 0.03, respectively). CONCLUSION: Neutrophil-to-lymphocyte ratio was related to pro-inflammatory, adiposity biomarkers and progressive subclinical atherogenesis.


Assuntos
Adipocinas/metabolismo , Aterosclerose/etiologia , Citocinas/metabolismo , Adiposidade/fisiologia , Adulto , Aterosclerose/sangue , Aterosclerose/patologia , Biomarcadores/metabolismo , Espessura Intima-Media Carotídea , Progressão da Doença , Feminino , Humanos , Gordura Intra-Abdominal/diagnóstico por imagem , Linfócitos/fisiologia , Masculino , Pessoa de Meia-Idade , Neutrófilos/fisiologia , Obesidade/sangue , Obesidade/patologia , Estudos Prospectivos
3.
J Infect Dev Ctries ; 17(5): 707-712, 2023 05 31.
Artigo em Inglês | MEDLINE | ID: mdl-37279416

RESUMO

INTRODUCTION: Infantile acute gastroenteritis (AGE) is a leading cause of morbidity and mortality, particularly in developing countries. The most frequent etiological agents of viral gastroenteritis in children are adenovirus, astrovirus, rotavirus, and norovirus, the last two, leading causes. Thus, the aim of this study was to identify the presence of these two viruses in children with AGE, from two cities located in the Southeast and the Northwest regions of México. METHODOLOGY: HuNoVs were detected and characterized by RT-PCR and sequencing, while RVs were detected by RNA electrophoresis. RESULTS: The presence of RV and HuNoV was evaluated in 81 stool samples; 37 were collected between April and July 2013 from patients with acute diarrhea in Merida, and 44 were collected between January and June 2017 in Chihuahua, who attended health services. Despite vaccination, RV resulted in the predominant viruses detected, with 30.8% (25/81) positivity, while HuNoV infection was present in 8.6% (7/81) of the stool samples; GII strains were identified circulating in the Southeast, while GI strains were identified in the Northwest. Moreover, co-infections with both viruses were detected at a prevalence rate of 2.4% (2/81). CONCLUSIONS: The circulation of RV and HuNoV in the country is continuous and should be constantly monitored due to their impact on public health.


Assuntos
Infecções por Caliciviridae , Gastroenterite , Norovirus , Infecções por Rotavirus , Rotavirus , Vírus , Humanos , Criança , Lactente , Rotavirus/genética , Norovirus/genética , Cidades , México/epidemiologia , Gastroenterite/epidemiologia , Vírus/genética , Fezes , Infecções por Caliciviridae/epidemiologia , Infecções por Rotavirus/epidemiologia
4.
J Gen Virol ; 91(Pt 11): 2713-8, 2010 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-20631089

RESUMO

Replicon systems have been useful to study mechanisms of translation and replication of flavivirus RNAs. In this study, we constructed a dengue virus 4 replicon encoding a Renilla luciferase (R(luc)) reporter, and six single-residue substitution mutants were generated: L128F and S158P in the non-structural protein (NS) 3 protease domain gene, and N96I, N390A, K437R and M805I in the NS5 gene. The effects of these substitutions on viral RNA translation and/or replication were examined by measuring R(luc) activities in wild-type and mutant replicon RNA-transfected Vero cells incubated at 35, 37 and 39 °C. Our results show that none of the mutations affected translation of replicon RNAs; however, L128F and S158P of NS3 at 39°C, and N96I of NS5 at 37 and 39°C, presented temperature-sensitive (ts) phenotypes for replication. Furthermore, using in vitro methyltransferase assays, we identified that the N96I mutation in NS5 exhibited a ts phenotype for N7-methylation, but not for 2'-O-methylation.


Assuntos
Vírus da Dengue/fisiologia , Mutação de Sentido Incorreto , Biossíntese de Proteínas , Replicon , Temperatura , Proteínas não Estruturais Virais/genética , Replicação Viral , Substituição de Aminoácidos/genética , Animais , Chlorocebus aethiops , Vírus da Dengue/genética , Vírus da Dengue/crescimento & desenvolvimento , Genes Essenciais , Genes Reporter , Genes Virais , Luciferases de Renilla/genética , Luciferases de Renilla/metabolismo , Estabilidade Proteica , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Células Vero
5.
Viruses ; 12(7)2020 07 16.
Artigo em Inglês | MEDLINE | ID: mdl-32708685

RESUMO

The arthropod-borne flaviviruses are important human pathogens, and a deeper understanding of the virus-host cell interaction is required to identify cellular targets that can be used as therapeutic candidates. It is well reported that the flaviviruses hijack several cellular functions, such as exosome-mediated cell communication during infection, which is modulated by the delivery of the exosomal cargo of pro- or antiviral molecules to the receiving host cells. Therefore, to study the role of exosomes during flavivirus infections is essential, not only to understand its relevance in virus-host interaction, but also to identify molecular factors that may contribute to the development of new strategies to block these viral infections. This review explores the implications of exosomes in flavivirus dissemination and transmission from the vector to human host cells, as well as their involvement in the host immune response. The hypothesis about exosomes as a transplacental infection route of ZIKV and the paradox effect or the dual role of exosomes released during flavivirus infection are also discussed here. Although several studies have been performed in order to identify and characterize cellular and viral molecules released in exosomes, it is not clear how all of these components participate in viral pathogenesis. Further studies will determine the balance between protective and harmful exosomes secreted by flavivirus infected cells, the characteristics and components that distinguish them both, and how they could be a factor that determines the infection outcome.


Assuntos
Comunicação Celular , Exossomos/metabolismo , Infecções por Flavivirus/metabolismo , Flavivirus/metabolismo , Interações Hospedeiro-Patógeno , Animais , Vetores Aracnídeos/virologia , Dengue/metabolismo , Vesículas Extracelulares/metabolismo , Vesículas Extracelulares/virologia , Infecções por Flavivirus/transmissão , Humanos , Mosquitos Vetores/virologia , Carrapatos/virologia , Infecção por Zika virus/metabolismo
6.
Infect Genet Evol ; 71: 98-107, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30905775

RESUMO

Dengue virus is the most prevalent arbovirus in Mexico, and although the diversity of this virus has been studied, the vast majority of sequences have been derived from viruses isolated from the human host. In this work, we aimed to sequence and to analyze DENVs derived from wild mosquitoes captured in Acapulco Guerrero, Mexico. We succeeded in determining three full genome sequences of such viruses and were able to compare them with other reported sequences from human and mosquito-derived DENVs. We found 15 nonsynonymous and 88 synonymous substitutions that were present more frequently in mosquito viruses than what would be expected by chance, although the limited number of genomes reported so far puts a constraint on the conclusions that can be derived from these analyses. Also, given the high depth of coverage attained in one of the genomes a variant analysis was carried out, finding 68 polymorphic sites in this genome. Interestingly, six of them corresponded to SNV that were detected as potentially differential between mosquitoes and humans, indicating that a that at least some positions may be maintained as polymorphic, which may facilitate host transmission.


Assuntos
Aedes/virologia , Vírus da Dengue/genética , Dengue/transmissão , Mosquitos Vetores/virologia , Animais , Genoma Viral/genética , Técnicas de Genotipagem , Humanos , México , Filogenia , Polimorfismo Genético , Sequenciamento Completo do Genoma
7.
Carbohydr Polym ; 223: 115113, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31427016

RESUMO

We present three easily rationalized star-shaped PEGylated ß-cyclodextrin (ßCD) derivatives synthesized via conjugation of different molecular weight PEG chains (5000, 2000, and 550 Da) to the ßCD primary face by click chemistry (ßCD-PEG5000, ßCD-PEG2000, ßCD-PEG550 respectively). ßCDPEG systems are envisioned to further carry bioactive molecules, therefore, their interactions with biological interfaces must be determined at an early stage of development. Hence, the effect of ßCDPEGs chain length on cell viability was investigated. To this aim, three models were selected: Vero cells for their fibroblast-like features; HeLa cells that are commonly used for preliminary viability screening; and human peripheral monocytes which are macrophage precursors. Of the three pegylated derivatives, ßCD-PEG550 was the one that significantly affected HeLa cells and human monocytes viability. Despite the popularity of PEGylation approach, our results underscore the importance of careful and systematic PEGylated materials design for their future success in drug delivery systems.


Assuntos
Química Click , Monócitos/efeitos dos fármacos , Polietilenoglicóis/química , beta-Ciclodextrinas/farmacologia , Animais , Sobrevivência Celular/efeitos dos fármacos , Chlorocebus aethiops , Células HeLa , Humanos , Estrutura Molecular , Tamanho da Partícula , Propriedades de Superfície , Células Vero , beta-Ciclodextrinas/síntese química , beta-Ciclodextrinas/química
8.
Arch Oral Biol ; 87: 211-217, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29310033

RESUMO

OBJECTIVE: Epigenetic mechanisms, such as DNA methylation, regulate important biological processes as gene expression and it was suggested that these phenomena play important roles in the carcinogenesis and tumor biology. The aim of this review is to provide the current state of knowledge about epigenetic alterations, focusing mainly on DNA methylation, reported in odontogenic tumors. DESIGN: Literatures were searched based in the combination of the following keywords: odontogenic tumors, epigenetics, DNA methylation, histone modifications, non-coding RNA, microRNA, DNA methyltransferases. Electronic databases (Medline/PubMed, Scopus and Web of Science) were screened. RESULTS: The analysis of epigenetic alterations in different tumors has rapidly increased; however, limited information is available about epigenetic mechanisms involved in the formation of odontogenic tumors. DNA methylation is the most studied epigenetic modification in these tumors and the participation of non-coding RNA's in odontogenic tumors has been recently addressed. Differential expression of DNA methyltransferases, altered DNA methylation patterns and aberrant expression of non-coding RNA's were reported in odontogenic tumors. CONCLUSIONS: Current studies suggest epigenetics as an emerging mechanism, possibly implicated in etiopathogenesis of odontogenic tumors. Deeper understanding of the epigenetic abnormalities in these tumors could show potential applications as biomarkers or therapeutic possibilities in the future.


Assuntos
Metilação de DNA , Epigênese Genética , Tumores Odontogênicos/genética , Expressão Gênica , Humanos
9.
Carbohydr Polym ; 184: 20-29, 2018 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-29352912

RESUMO

In spite of the progress in the cyclodextrins chemistry, the synthesis of monodisperse derivatives with a defined degree of substitution is still a challenge. In this work we present a novel dendritic material produced by combining ßCD and second generation poly(ester) dendrons. The selective attachment of dendrons in the seven positions of the ßCD-primary face was performed through a CuAAC click reaction, which along with a very simple work-up, allowed obtaining the monodisperse material in very high yields. The product showed a great aqueous solubility and an in vitro non-toxic profile. The enhanced complexation potential of the product was evidenced through the formation of an inclusion complex with albendazole, which presented a Kc = 29636.17 M-1. In this system, albendazole was 45 times more water-soluble in comparison to the complex albendazole-native ßCD. All these features make the dendritic material very attractive for further applications in the formulation and drug delivery fields.

10.
J Int Med Res ; 45(6): 1879-1891, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27503947

RESUMO

Objective To evaluate the relationship between pro-atherogenic biomarkers and epicardial adipose tissue (EAT) thickness in patients with cardiovascular risk factors. Methods Plasma nitric oxide (NO), soluble intercellular adhesion molecule-1 and malondialdehyde (MDA) levels, EAT thickness, flow-mediated dilation (FMD) and carotid intima media thickness (CIMT) were determined in patients aged >18 years who were referred for echocardiography for heart ischemia or non-ischemic diseases. Cardiovascular risk factors (Framingham score [FS] ≥ 20) were weighted. Results Hypertension, dyslipidaemia and type 2 diabetes mellitus were prevalent (≥55% of 40 patients). Patients with FS ≥ 20 ( n = 21) showed significantly higher EAT and CIMT values. Globally, MDA, CIMT, age, waist circumference, high-density lipoprotein cholesterol (HDL-C) and FS were associated with EAT thickness. EAT was significantly associated with NO in patients with FS ≥ 20. Significant differences in EAT thickness were found between patients stratified by NO value, FMD, age, smoking status, dyslipidaemia, type 2 diabetes mellitus and FS. An EAT-associated atherogenic risk (CIMT ≥ 1 mm) model was statistically significant when MDA and type 2 diabetes mellitus were included. Conclusion EAT thickness was associated with MDA, CIMT, age, waist circumference, HDL-C and FS globally, but with NO only in patients with FS≥20. EAT may be used to identify vascular damage stage, possibly influenced by MDA and type 2 diabetes mellitus.


Assuntos
Tecido Adiposo/patologia , Aterosclerose/patologia , Espessura Intima-Media Carotídea , Pericárdio/patologia , Tecido Adiposo/diagnóstico por imagem , Adulto , Idoso , Idoso de 80 Anos ou mais , Aterosclerose/diagnóstico por imagem , Biomarcadores/metabolismo , Demografia , Feminino , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Pericárdio/diagnóstico por imagem , Fatores de Risco
11.
Genome Announc ; 5(12)2017 Mar 23.
Artigo em Inglês | MEDLINE | ID: mdl-28336600

RESUMO

Zika virus (ZIKV) is an emerging arthropod-borne flavivirus associated with severe congenital malformations and neurological complications. Although the ZIKV genome is well characterized, there is limited information regarding changes after cell isolation and culture adaptation. We isolated, and passaged in Vero cells, ZIKV from the serum of a symptomatic male patient and compared the viral genomes before and after culture. Single nucleotide polymorphisms were characteristic among serum-circulating genomes, while such diversity decreased after cell culture.

12.
Nat Genet ; 49(1): 36-45, 2017 01.
Artigo em Inglês | MEDLINE | ID: mdl-27841880

RESUMO

Despite the clinical significance of balanced chromosomal abnormalities (BCAs), their characterization has largely been restricted to cytogenetic resolution. We explored the landscape of BCAs at nucleotide resolution in 273 subjects with a spectrum of congenital anomalies. Whole-genome sequencing revised 93% of karyotypes and demonstrated complexity that was cryptic to karyotyping in 21% of BCAs, highlighting the limitations of conventional cytogenetic approaches. At least 33.9% of BCAs resulted in gene disruption that likely contributed to the developmental phenotype, 5.2% were associated with pathogenic genomic imbalances, and 7.3% disrupted topologically associated domains (TADs) encompassing known syndromic loci. Remarkably, BCA breakpoints in eight subjects altered a single TAD encompassing MEF2C, a known driver of 5q14.3 microdeletion syndrome, resulting in decreased MEF2C expression. We propose that sequence-level resolution dramatically improves prediction of clinical outcomes for balanced rearrangements and provides insight into new pathogenic mechanisms, such as altered regulation due to changes in chromosome topology.


Assuntos
Aberrações Cromossômicas , Anormalidades Congênitas/genética , Rearranjo Gênico , Marcadores Genéticos/genética , Predisposição Genética para Doença , Estudo de Associação Genômica Ampla , Feminino , Humanos , Masculino
13.
J Cancer ; 7(13): 1856-1860, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27698925

RESUMO

Chromatin in cervical cancer (CC) undergoes chemical and structural changes that alter the expression pattern of genes. Recently, a potential mechanism, which regulates gene expression at transcriptional levels is the proteolytic clipping of histone H3. However, until now this process in CC has not been reported. Using HeLa cells as a model of CC and human samples from patients with CC, we identify that the H3 cleavage was lower in CC compared with control tissue. Additionally, the histone H3 clipping was performed by serine and aspartyl proteases in HeLa cells. These results suggest that histone H3 clipping operates as part of post-translational modification system in CC.

14.
Virology ; 484: 113-126, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26092250

RESUMO

Given dengue virus (DENV) genome austerity, it uses cellular molecules and structures for virion entry, translation and replication of the genome. NS1 is a multifunctional protein key to viral replication and pathogenesis. Identification of cellular proteins that interact with NS1 may help in further understanding the functions of NS1. In this paper we isolated a total of 64 proteins from DENV infected human hepatic cells (Huh-7) that interact with NS1 by affinity chromatography and immunoprecipitation assays. The subcellular location and expression levels during infection of the ribosomal proteins RPS3a, RPL7, RPL18, RPL18a plus GAPDH were determined. None of these proteins changed their expression levels during infection; however, RPL-18 was redistributed to the perinuclear region after 48hpi. Silencing of the RPL-18 does not affect cell translation efficiency or viability, but it reduces significantly viral translation, replication and viral yield, suggesting that the RPL-18 is required during DENV replicative cycle.


Assuntos
Vírus da Dengue/fisiologia , Hepatócitos/virologia , Interações Hospedeiro-Patógeno , Biossíntese de Proteínas , Proteínas Ribossômicas/metabolismo , Proteínas não Estruturais Virais/metabolismo , Replicação Viral , Linhagem Celular , Cromatografia de Afinidade , Humanos , Imunoprecipitação , Ligação Proteica
15.
Methods Mol Biol ; 1138: 131-50, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24696335

RESUMO

Dengue virus serotypes 1-4 are members of mosquito-borne flavivirus genus of Flaviviridae family that encode one long open reading frame (ORF) that is translated to a polyprotein. Both host and virally encoded proteases function in the processing of the polyprotein by co-translational and posttranslational mechanisms to yield 10 mature proteins prior to viral RNA replication. To study cis- and trans-acting factors involved in viral RNA replication, many groups [1-8] have constructed cDNAs encoding West Nile virus (WNV), DENV, or yellow fever virus reporter replicon RNAs. The replicon plasmids constructed in our laboratory for WNV [9] and the DENV4 replicon described here are arranged in the order of 5'-untranslated region (UTR), the N-terminal coding sequence of capsid (C), Renilla luciferase (Rluc) reporter gene with a translation termination codon, and an internal ribosome entry site (IRES) element from encephalomyocarditis virus (EMCV) for cap-independent translation of the downstream ORF that codes for a polyprotein precursor, CterE-NS1-NS2A-NS2B-NS3-NS4A-NS4B-NS5, followed by the 3'-UTR. In the second DENV4 replicon, the Rluc gene is fused sequentially downstream to the 20 amino acid (aa) FMDV 2A protease coding sequence, neomycin resistance gene (Neo(r)), a termination codon, and the EMCV leader followed by the same polyprotein coding sequence and 3'-UTR as in the first replicon. The first replicon is useful to study by transient transfection experiments the cis-acting elements and trans-acting factors involved in viral RNA replication. The second DENV4 replicon is used to establish a stable monkey kidney (Vero) cell line by transfection of replicon RNA and selection in the presence of the G418, an analog of neomycin. This replicon is useful for screening and identifying antiviral compounds that are potential inhibitors of viral replication.


Assuntos
Vírus da Dengue/genética , Vírus da Dengue/fisiologia , Genoma Viral/genética , Biologia Molecular/métodos , Replicon/genética , Replicação Viral/fisiologia , Animais , Linhagem Celular , Clonagem Molecular , Eletroporação , Genes Reporter , Vetores Genéticos/metabolismo , Recombinação Homóloga/genética , Humanos , Luciferases de Renilla/metabolismo , Plasmídeos/metabolismo , Reação em Cadeia da Polimerase , Saccharomyces cerevisiae/metabolismo , Transcrição Gênica , Transformação Genética
16.
Methods Mol Biol ; 1030: 283-99, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23821276

RESUMO

Mosquito-borne flavivirus RNA genomes encode one long open reading frame flanking 5'- and 3'-untranslated regions (5'- and 3'-UTRs) which contain cis-acting RNA elements playing important roles for viral RNA translation and replication. The viral RNA encodes a single polyprotein, which is processed into three structural proteins and seven nonstructural (NS) proteins. The regions coding for the seven NS proteins are sufficient for replication of the RNA. The sequences encoding the structural genes can be deleted except for two short regions. The first one encompasses 32 amino acid (aa) residues from the N-terminal coding sequence of capsid (C) and the second, 27 aa region from the C-terminus of envelope (E) protein. The deleted region can be substituted with a gene coding for a readily quantifiable reporter to give rise to a subgenomic reporter replicon. Replicons containing a variety of reporter genes and marker genes for construction of stable mammalian cell lines are valuable reagents for studying the effects of mutations in translation and/or replication in isolation from processes like the entry and assembly of the virus particles. Here we describe the construction of two West Nile virus (WNV) replicons by overlap extension PCR and standard recombinant DNA techniques. One has a Renilla luciferase (Rluc) reporter gene followed by an internal ribosome entry site (element) for cap-independent translation of the open reading frame encompassing the carboxy-terminal sequence of E to NS5. The second replicon has in tandem the Rluc gene, foot and mouth disease virus 2A, and neomycin phosphotransferase gene that allows establishment of a stable mammalian cell line expressing the Rluc reporter in the presence of the neomycin analog, G418. The stable replicon-expressing Vero cell line has been used for cell-based screening and determination of EC50 values for antiviral compounds that inhibited WNV replication.


Assuntos
Antivirais/farmacologia , Genoma Viral , Testes de Sensibilidade Microbiana/métodos , Replicação Viral/efeitos dos fármacos , Replicação Viral/genética , Vírus do Nilo Ocidental/efeitos dos fármacos , Vírus do Nilo Ocidental/fisiologia , Animais , Linhagem Celular , Chlorocebus aethiops , Clonagem Molecular , Expressão Gênica , Ordem dos Genes , Genes Reporter , Vetores Genéticos/genética , Transfecção , Células Vero
17.
Arch Oral Biol ; 87: 211-217, 2018.
Artigo em Inglês | URUCAN | ID: bcc-5169

RESUMO

OBJECTIVE:Epigenetic mechanisms, such as DNA methylation, regulate important biological processes as gene expression and it was suggested that these phenomena play important roles in the carcinogenesis and tumor biology. The aim of this review is to provide the current state of knowledge about epigenetic alterations, focusing mainly on DNA methylation, reported in odontogenic tumors.DESIGN:Literatures were searched based in the combination of the following keywords: odontogenic tumors, epigenetics, DNA methylation, histone modifications, non-coding RNA, microRNA, DNA methyltransferases. Electronic databases (Medline/PubMed, Scopus and Web of Science) were screened.RESULTS:The analysis of epigenetic alterations in different tumors has rapidly increased; however, limited information is available about epigenetic mechanisms involved in the formation of odontogenic tumors. DNA methylation is the most studied epigenetic modification in these tumors and the participation of non-coding RNA's in odontogenic tumors has been recently addressed. Differential expression of DNA methyltransferases, altered DNA methylation patterns and aberrant expression of non-coding RNA's were reported in odontogenic tumors.CONCLUSIONS:Current studies suggest epigenetics as an emerging mechanism, possibly implicated in etiopathogenesis of odontogenic tumors. Deeper understanding of the epigenetic abnormalities in these tumors could show potential applications as biomarkers or therapeutic possibilities in the future(AU)


Assuntos
Humanos , Tumores Odontogênicos/genética , Bibliografia Nacional , Uruguai
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