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1.
Front Public Health ; 8: 562615, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33072699

RESUMO

The pandemic caused by the new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a worldwide public health concern. First confined in China and then disseminated widely across Europe and America, SARS-CoV-2 has impacted and moved the scientific community around the world to working in a fast and coordinated way to collect all possible information about this virus and generate new strategies and protocols to try to stop the infection. During March 2020, more than 16,000 full viral genomes have been shared in public databases that allow the construction of genetic landscapes for tracking and monitoring the viral advances over time and study the genomic variations present in geographic regions. In this work, we present the occurrence of genetic variants and lineages of SARS-CoV-2 in Chile during March to April 2020. Complete genome analysis of 141 viral samples from different regions of Chile revealed a predominance of variant D614G like in Europe and the USA and the major presence of lineage B.1. These findings could help take control measures due to the similarity of the viral variants present in Chile, compared with other countries, and monitor the dynamic change of virus variants in the country.

2.
J Med Virol ; 92(9): 1562-1566, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32222995

RESUMO

The current pandemic caused by the new coronavirus is a worldwide public health concern. To aboard this emergency, and like never before, scientific groups around the world have been working in a fast and coordinated way to get the maximum of information about this virus when it has been almost 3 months since the first cases were detected in Wuhan province in China. The complete genome sequences of around 450 isolates are available, and studies about similarities and differences among them and with the close related viruses that caused similar epidemics in this century. In this work, we studied the complete genome of the first four cases of the new coronavirus disease in Chile, from patients who traveled to Europe and Southeast Asia. Our findings reveal at least two different viral variants entries to Chilean territory, coming from Europe and Asia. We also sub-classified the isolates into variants according to punctual mutations in the genome. Our work contributes to global information about transmission dynamics and the importance to take control measures to stop the spread of the infection.

3.
Rev Chilena Infectol ; 36(5): 585-590, 2019 Oct.
Artigo em Espanhol | MEDLINE | ID: mdl-31859799

RESUMO

BACKGROUND: Listeria monocytogenes is a foodborne pathogen that causes listeriosis, a disease that can present as febrile gastroenteritis or as an invasive form that has high mortality rates. So far, the genetic diversity of strains of L. monocytogenes isolated from patients, foods and environmental sources in Chile has been poorly studied. AIM: To characterize genetically L. monocytogenes strains received by the Institute of Public Health of Chile (ISP) between 2007 and 2014. METHODS: We selected 94 strains of L. monocytogenes corresponding to 94 different pulsotypes identified by pulsed field gel electrophoresis (PFGE), DNA was extracted and serotyping was performed by polymerase chain reaction (PCR) and multilocus sequence typing (MLST). RESULTS: The most common serotype was 4b (55.3%), followed by serotypes 1/2a (25.5%), 1/2b (17%) and 1/2c (2.2%). 32 sequence-type (ST) were identified, of which 4 were new, and the predominant ones were ST1 (28.7%) and ST2 (13.8%). All the strains of L. monocytogenes were grouped in Lineages I and II. CONCLUSIONS: A great genetic variability was observed in the strains of L. monocytogenes analyzed, being predominant the ST1 and ST2, both belonging to Lineage I. Our results contribute to know the population structure of this pathogen in Chile and its presence in clinical samples, food and the environment.


Assuntos
Listeria monocytogenes/genética , Listeria monocytogenes/isolamento & purificação , Chile , Eletroforese em Gel de Campo Pulsado , Microbiologia Ambiental , Microbiologia de Alimentos , Variação Genética , Humanos , Listeriose/microbiologia , Tipagem de Sequências Multilocus , Reação em Cadeia da Polimerase , Sorotipagem , Fatores de Tempo
4.
Rev. chil. infectol ; 36(5): 585-590, oct. 2019. graf
Artigo em Espanhol | LILACS-Express | LILACS | ID: biblio-1058084

RESUMO

Resumen Introducción: Listeria monocytogenes es un patógeno transmitido por alimentos que causa listeriosis, una enfermedad que puede presentarse como gastroenteritis febril o en una forma invasora que tiene altas tasas de mortalidad. Hasta el momento, ha sido poco estudiada la diversidad genética de cepas de L. monocytogenes aisladas desde pacientes, alimentos y fuentes ambientales en Chile. Objetivo: Caracterizar genéticamente cepas de L. monocytogenes de estos tres orígenes recibidas por el Instituto de Salud Pública de Chile (ISP) entre los años 2007 y 2014. Material y Métodos: Se seleccionaron 94 cepas de L. monocytogenes correspondientes a 94 pulsotipos diferentes identificados por electroforesis en gel de campo pulsado (PFGE), se extrajo ADN y se realizó serotipificación mediante reacción de polimerasa en cadena (RPC) y tipificación de secuencias multilocus (MLST). Resultados: El serotipo más común fue 4b (55,3%), seguido de 1/2a (25,5%), 1/2b (17%) y 1/2c (2,2%). Se identificaron 32 secuencias tipo (ST), de las cuales cuatro fueron nuevas, y las predominantes fueron ST1 (28,7%) y ST2 (13,8%). La totalidad de las cepas se agrupó en los Linajes I y II. Conclusiones: Se observó una gran variabilidad genética en las cepas de L. monocytogenes analizadas, siendo predominantes las secuencias tipo ST1 y ST2, ambas pertenecientes al Linaje I. Nuestros resultados contribuyen a conocer la estructura poblacional de este patógeno en Chile y su presencia en muestras clínicas, alimentos y el medio ambiente.


Background: Listeria monocytogenes is a foodborne pathogen that causes listeriosis, a disease that can present as febrile gastroenteritis or as an invasive form that has high mortality rates. So far, the genetic diversity of strains of L. monocytogenes isolated from patients, foods and environmental sources in Chile has been poorly studied. Aim: To characterize genetically L. monocytogenes strains received by the Institute of Public Health of Chile (ISP) between 2007 and 2014. Methods: We selected 94 strains of L. monocytogenes corresponding to 94 different pulsotypes identified by pulsed field gel electrophoresis (PFGE), DNA was extracted and serotyping was performed by polymerase chain reaction (PCR) and multilocus sequence typing (MLST). Results: The most common serotype was 4b (55.3%), followed by serotypes 1/2a (25.5%), 1/2b (17%) and 1/2c (2.2%). 32 sequence-type (ST) were identified, of which 4 were new, and the predominant ones were ST1 (28.7%) and ST2 (13.8%). All the strains of L. monocytogenes were grouped in Lineages I and II. Conclusions: A great genetic variability was observed in the strains of L. monocytogenes analyzed, being predominant the ST1 and ST2, both belonging to Lineage I. Our results contribute to know the population structure of this pathogen in Chile and its presence in clinical samples, food and the environment.

5.
Med Microbiol Immunol ; 208(6): 757-771, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31165237

RESUMO

The high-risk human papillomaviruses (HR-HPVs) are involved in the development of cervical cancer. Nevertheless, there are differences in the oncogenic potential among them. HPV-16 and HPV-18 are associated with approximately 70% of cancer worldwide, and both types are the most extensively studied HR-HPV. Great variations in the prevalence of HR-HPV have been described in different countries. The impact of these variations on the epidemiology of lesions and cervical cancer is currently unknown. A high prevalence of HPV-66 has been detected in Chile. Here, we have analyzed the genetic variability of the L1 gene from HPV-66-infected Chilean women. Higher order interactions between identified mutations were analyzed by co-variation and cluster analyses. Antigenic-index alterations following L1 mutations and B-cell epitopes were predicted by BcePred algorithm. HPV-66 L1 sequences clustered phylogenetically into two main clades. The genetic variability in the HPV-66 L1 gene involved thirty nucleotide changes. Four of these were for the first time identified in this study. Some of these variants are embedded in the B-cell epitope regions. Amino acid homology in the immunodominant epitopes of HPV-66 L1 protein (DE, FG and H1 loops) was 42.9-59.1% and 28.6-68.9% compared with HPV-16 and HPV-18, respectively. The results of this research suggest that the neutralizing epitopes of HPV-66 are antigenically different compared to HPV-16 and HPV-18. Our findings show the need to perform new structural and immunological studies on HPV-66 L1 protein to evaluate the cross-protection conferred by current HPV vaccines.


Assuntos
Proteínas do Capsídeo/genética , Detecção Precoce de Câncer , Variação Genética , Genótipo , Proteínas Oncogênicas Virais/genética , Papillomaviridae/genética , Neoplasias do Colo do Útero/epidemiologia , Adolescente , Adulto , Antígenos Virais/genética , Chile/epidemiologia , Epitopos de Linfócito B/genética , Feminino , Humanos , Pessoa de Meia-Idade , Papillomaviridae/isolamento & purificação , Prevalência , Neoplasias do Colo do Útero/diagnóstico , Neoplasias do Colo do Útero/virologia , Adulto Jovem
6.
PLoS One ; 11(8): e0160434, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27518286

RESUMO

Tuberculosis (TB), caused by the pathogen Mycobacterium tuberculosis (MTB), remains a disease of high importance to global public health. Studies into the population structure of MTB have become vital to monitoring possible outbreaks and also to develop strategies regarding disease control. Although Chile has a low incidence of MTB, the current rates of migration have the potential to change this scenario. We collected and analyzed a total of 458 M. tuberculosis isolates (1 isolate per patient) originating from all 15 regions of Chile. The isolates were genotyped using the spoligotyping method and the data obtained were analyzed and compared with the SITVIT2 database. A total of 169 different patterns were identified, of which, 119 patterns (408 strains) corresponded to Spoligotype International Types (SITs) and 50 patterns corresponded to orphan strains. The most abundantly represented SITs/lineages were: SIT53/T1 (11.57%), SIT33/LAM3 (9.6%), SIT42/LAM9 (9.39%), SIT50/H3 (5.9%), SIT37/T3 (5%); analysis of the spoligotyping minimum spanning tree as well as spoligoforest were suggestive of a recent expansion of SIT42, SIT50 and SIT37; all of which potentially evolved from SIT53. The most abundantly represented lineages were LAM (40.6%), T (34.1%) and Haarlem (13.5%). LAM was more prevalent in the Santiago (43.6%) and Concepción (44.1%) isolates, rather than the Iquique (29.4%) strains. The proportion of X lineage was appreciably higher in Iquique and Concepción (11.7% in both) as compared to Santiago (1.6%). Global analysis of MTB lineage distribution in Chile versus neighboring countries showed that evolutionary recent lineages (LAM, T and Haarlem) accounted together for 88.2% of isolates in Chile, a pattern which mirrored MTB lineage distribution in neighboring countries (n = 7378 isolates recorded in SITVIT2 database for Peru, Brazil, Paraguay, and Argentina; and published studies), highlighting epidemiological advantage of Euro-American lineages in this region. Finally, we also observed exclusive emergence of patterns SIT4014/X1 and SIT4015 (unknown lineage signature) that have hitherto been found exclusively in Chile, indicating that conditions specific to Chile, along with the unique genetic makeup of the Chilean population, might have allowed for a possible co-evolution leading to the success of these emerging genotypes.


Assuntos
Técnicas de Tipagem Bacteriana/métodos , Variação Genética/genética , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/isolamento & purificação , Tuberculose/epidemiologia , Argentina/epidemiologia , Brasil/epidemiologia , Chile/epidemiologia , Genótipo , Humanos , Incidência , Mycobacterium tuberculosis/classificação , Paraguai/epidemiologia , Peru/epidemiologia , Filogenia , Tuberculose/microbiologia
7.
J Proteome Res ; 15(5): 1659-69, 2016 05 06.
Artigo em Inglês | MEDLINE | ID: mdl-27049334

RESUMO

Nucleolin (NCL) is a major component of the cell nucleolus, which has the ability to rapidly shuttle to several other cells' compartments. NCL plays important roles in a variety of essential functions, among which are ribosome biogenesis, gene expression, and cell growth. However, the precise mechanisms underlying NCL functions are still unclear. Our study aimed to provide new information on NCL functions via the identification of its nuclear interacting partners. Using an interactomics approach, we identified 140 proteins co-purified with NCL, among which 100 of them were specifically found to be associated with NCL after RNase digestion. The functional classification of these proteins confirmed the prominent role of NCL in ribosome biogenesis and additionally revealed the possible involvement of nuclear NCL in several pre-mRNA processing pathways through its interaction with RNA helicases and proteins participating in pre-mRNA splicing, transport, or stability. NCL knockdown experiments revealed that NCL regulates the localization of EXOSC10 and the amount of ZC3HAV1, two components of the RNA exosome, further suggesting its involvement in the control of mRNA stability. Altogether, this study describes the first nuclear interactome of human NCL and provides the basis for further understanding the mechanisms underlying the essential functions of this nucleolar protein.


Assuntos
Fosfoproteínas/fisiologia , Proteômica/métodos , Proteínas de Ligação a RNA/fisiologia , Complexo Multienzimático de Ribonucleases do Exossomo/química , Humanos , Proteínas Nucleares/metabolismo , Fosfoproteínas/metabolismo , Ligação Proteica , RNA Helicases/metabolismo , Estabilidade de RNA , Proteínas de Ligação a RNA/metabolismo , Ribossomos
8.
Rev. chil. neuro-psiquiatr ; 53(1): 53-58, mar. 2015. ilus
Artigo em Espanhol | LILACS | ID: lil-745588

RESUMO

Microglial cells play a major role in the innate immunity of the central nervous system. Alterations in the normal cross-talks between microglia and brain neuronal cells may lead to serious disturbances and neurodegenerative diseases. We have postulated that neuroinflammatory processes are a critical factor triggering the pathological cascade leading to neuronal degeneration. In our neuroimmunomodulation theory, external or internal damage signals activate microglial cells, producing cytotoxic factors that induce neuronal degeneration. These factors activate protein-kinases, that lead to tau hyperphosphorylation, and its consequent oligomerization. The tau aggregates released into the extracellular medium favor a positive feedback mechanism that determines neurodegeneration. Nowadays, natural components with a string anti-inflammatory activity and that cross the blood brain barrier appears as candidates for prevention and treatment of degenerative brain disorders such as Alzheimers'disease.


Las células microgliales juegan un papel importante en la inmunidad innata del sistema nervioso central. Las alteraciones en la normal diafonía celular, entre microglias y células neuronales cerebrales, pueden conducir a graves disturbios y enfermedades neurodegenerativas. En este contexto, hemos postulado que los procesos neuroinflamatorios son un factor crítico a desencadenar la cascada patológica que conduce a la degeneración neuronal. En nuestra teoría Neuroinmunomoduladora, señales de daños externos o internos activan las células microgliales, favoreciendo la producción de factores citotóxicos que inducen la degeneración neuronal. Estos factores activan la proteína-quinasas, que conducen a la hiperfosforilación de la proteína tau, y su consecuente oligomerización. Estos agregados de tau liberados al medio extracelular, al activar a la célula microglial, provocarían un mecanismo de retroalimentación positiva favoreciendo la neurodegeneración. Hoy en día, compuestos de origen natural con una fuerte actividad anti-inflamatoria, capaces de cruzar la barrera hematoencefálica del cerebro, aparecen como candidatos para la prevención y el tratamiento de trastornos neurodegenerativos tales como la enfermedad de Alzheimer.


Assuntos
Humanos , Terapêutica , Neuroimunomodulação , Doenças Neurodegenerativas
9.
J Virol ; 86(3): 1449-57, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22130536

RESUMO

Herpes simplex virus type 1 (HSV-1) infection induces profound nucleolar modifications at the functional and organizational levels, including nucleolar invasion by several viral proteins. One of these proteins is US11, which exhibits several different functions and displays both cytoplasmic localization and clear nucleolar localization very similar to that of the major multifunctional nucleolar protein nucleolin. To determine whether US11 interacts with nucleolin, we purified US11 protein partners by coimmunoprecipitations using a tagged protein, Flag-US11. From extracts of cells expressing Flag-US11 protein, we copurified a protein of about 100 kDa that was further identified as nucleolin. In vitro studies have demonstrated that nucleolin interacts with US11 and that the C-terminal domain of US11, which is required for US11 nucleolar accumulation, is sufficient for interaction with nucleolin. This association was confirmed in HSV-1-infected cells. We found an increase in the nucleolar accumulation of US11 in nucleolin-depleted cells, thereby revealing that nucleolin could play a role in US11 nucleocytoplasmic trafficking through one-way directional transport out of the nucleolus. Since nucleolin is required for HSV-1 nuclear egress, the interaction of US11 with nucleolin may participate in the outcome of infection.


Assuntos
Fosfoproteínas/metabolismo , Proteínas de Ligação a RNA/metabolismo , Proteínas Virais/metabolismo , Far-Western Blotting , Células HeLa , Humanos , Imunoprecipitação , Ligação Proteica , Transporte Proteico , RNA Interferente Pequeno
10.
J Virol ; 84(17): 8871-87, 2010 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-20573815

RESUMO

Adeno-associated virus (AAV) is a human parvovirus that replicates only in cells coinfected with a helper virus, such as adenovirus or herpes simplex virus type 1 (HSV-1). We previously showed that nine HSV-1 factors are able to support AAV rep gene expression and genome replication. To elucidate the strategy of AAV replication in the presence of HSV-1, we undertook a proteomic analysis of cellular and HSV-1 factors associated with Rep proteins and thus potentially recruited within AAV replication compartments (AAV RCs). This study resulted in the identification of approximately 60 cellular proteins, among which factors involved in DNA and RNA metabolism represented the largest functional categories. Validation analyses indicated that the cellular DNA replication enzymes RPA, RFC, and PCNA were recruited within HSV-1-induced AAV RCs. Polymerase delta was not identified but subsequently was shown to colocalize with Rep within AAV RCs even in the presence of the HSV-1 polymerase complex. In addition, we found that AAV replication is associated with the recruitment of components of the Mre11/Rad50/Nbs1 complex, Ku70 and -86, and the mismatch repair proteins MSH2, -3, and -6. Finally, several HSV-1 factors were also found to be associated with Rep, including UL12. We demonstrated for the first time that this protein plays a role during AAV replication by enhancing the resolution of AAV replicative forms and AAV particle production. Altogether, these analyses provide the basis to understand how AAV adapts its replication strategy to the nuclear environment induced by the helper virus.


Assuntos
Proteínas de Ligação a DNA/metabolismo , Dependovirus/fisiologia , Herpes Simples/metabolismo , Herpesvirus Humano 1/fisiologia , Infecções por Parvoviridae/metabolismo , Proteínas Virais/metabolismo , Replicação Viral , Animais , Chlorocebus aethiops , Replicação do DNA , Proteínas de Ligação a DNA/genética , Dependovirus/genética , Células HeLa , Herpes Simples/virologia , Herpesvirus Humano 1/genética , Humanos , Infecções por Parvoviridae/virologia , Ligação Proteica , Mapeamento de Interação de Proteínas , Proteína de Replicação A/genética , Proteína de Replicação A/metabolismo , Proteína de Replicação C/genética , Proteína de Replicação C/metabolismo , Células Vero , Proteínas Virais/genética
11.
Vet Microbiol ; 141(1-2): 53-8, 2010 Feb 24.
Artigo em Inglês | MEDLINE | ID: mdl-19818567

RESUMO

Infectious pancreatic necrosis is a disease caused by a birnavirus affecting several wild and commercial aquatic organisms. This infectious disease results in significant losses in the farming industry and therefore effective therapeutic agents are needed to control outbreaks caused by this pathogen. Our goal was to evaluate in vitro antiviral effect of a group of natural compounds (geranyl aromatic derivatives) isolated from the resinous exudate of the plant Heliotropium filifolium (Heliotropiaceae), semi-synthetics compounds obtained from them, and the resinous exudate, on CHSE-214 cell line infected with infectious pancreatic necrosis virus (IPNV) using a virus plaque inhibition assay at various concentrations. The compound ester filifolinyl senecionate was the best antiviral with EC(50) 160 microg/mL and a cytotoxic concentration required to reduce cell viability by 50% up to 400 microg/mL. In order to obtain information about the mechanism of the antiviral action, was evaluated the influence of ester filifolinyl senecionate on the viral RNA synthesis. This compound produced inhibition of the synthesis of viral genomic RNA, suggesting that the ester could be interacting with the viral RNA during the viral cycle. Additionally, a preliminary study of the interaction between ester and a sample of single-stranded RNA was studied at the level of theory Restricted Hartree Fock PM3 method. The results showed that the ester formed hydrogen bonds mainly with nitrogenous bases but not with ribose and phosphate. These results allow propose that the ester filifolinyl senecionate is a good candidate for used as antiviral therapy for IPN virus in salmon fry.


Assuntos
Antivirais/farmacologia , Heliotropium/química , Vírus da Necrose Pancreática Infecciosa/efeitos dos fármacos , Vírus da Necrose Pancreática Infecciosa/fisiologia , Exsudatos de Plantas/farmacologia , Replicação Viral/efeitos dos fármacos , Animais , Antivirais/síntese química , Antivirais/química , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , DNA/metabolismo , Exsudatos de Plantas/química , RNA/metabolismo , Salmão
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