Quantitative proteomic analysis of Huh-7 cells infected with Dengue virus by label-free LC–MS

AbstractDengue is an important and growing public health problem worldwide with an estimated 100million new clinical cases annually. Currently, no licensed drug or vaccine is available. During natural infection in humans, liver cells constitute one of the main targets of dengue virus (DENV) replication. However, a clear understanding of dengue pathogenesis remains elusive. In order to gain a better reading of the cross talk between virus and host cell proteins, we used a proteomics approach to analyze the host response to DENV infection in a hepatic cell line Huh-7. Differences in proteome expression were assayed 24h post-infection using label-free LC–MS. Quantitative analysis revealed 155 differentially expressed proteins, 64 of which were up-regulated and 91 down-regulated. These results reveal an important decrease in the expression of enzymes involved in the glycolytic pathway, citrate cycle, and pyruvate metabolism. This study provides large-scale quantitative information regarding protein expression in the early stages of infection that should be useful for better compression of the pathogenesis of dengue.Biological significanceDengue infection involves alterations in the homeostasis of the host cell. Defining the interactions between virus and cell proteins should provide a better understanding of how viruses propagate and cause disease. Here, we present for the first time the proteomic analysis of hepatocytes (Huh-7 cells) infected with DENV-2 by label-free LC–MS.This article is part of a Special Issue entitled: Proteomics, mass spectrometry and peptidomics, Cancun 2013. Guest Editors: César López-Camarillo, Victoria Pando-Robles and Bronwyn Jane Barkla

Estimating mortality and disability in Peru before the COVID-19 pandemic: a systematic analysis from the Global Burden of the Disease Study 2019

"Background: Estimating and analyzing trends and patterns of health loss are essential to promote efficient resource allocation and improve Peru’s healthcare system performance. Methods: Using estimates from the Global Burden of Disease (GBD), Injuries, and Risk Factors Study (2019), we assessed mortality and disability in Peru from 1990 to 2019. We report demographic and epidemiologic trends in terms of population, life expectancy at birth (LE), mortality, incidence, prevalence, years of life lost (YLLs), years lived with disability (YLDs), and disability-adjusted life-years (DALYs) caused by the major diseases and risk factors in Peru. Finally, we compared Peru with 16 countries in the Latin American (LA) region. Results: The Peruvian population reached 33.9 million inhabitants (49.9% women) in 2019. From 1990 to 2019, LE at birth increased from 69.2 (95% uncertainty interval 67.8–70.3) to 80.3 (77.2–83.2) years. This increase was driven by the decline in under-5 mortality (−80.7%) and mortality from infectious diseases in older age groups (+60 years old). The number of DALYs in 1990 was 9.2 million (8.5–10.1) and reached 7.5 million (6.1–9.0) in 2019. The proportion of DALYs due to non-communicable diseases (NCDs) increased from 38.2% in 1990 to 67.9% in 2019. The all-ages and age-standardized DALYs rates and YLLs rates decreased, but YLDs rates remained constant. In 2019, the leading causes of DALYs were neonatal disorders, lower respiratory infections (LRIs), ischemic heart disease, road injuries, and low back pain. The leading risk factors associated with DALYs in 2019 were undernutrition, high body mass index, high fasting plasma glucose, and air pollution. Before the COVID-19 pandemic, Peru experienced one of the highest LRIs-DALYs rates in the LA region. Conclusion: In the last three decades, Peru experienced significant improvements in LE and child survival and an increase in the burden of NCDs and associated disability. The Peruvian healthcare system must be redesigned to respond to this epidemiological transition. The new design should aim to reduce premature deaths and maintain healthy longevity, focusing on effective coverage and treatment of NCDs and reducing and managing the related disability.

La importancia de la proteómica en la salud pública: The significance of proteomics in public health

La salud pública en el nuevo milenio tiene como reto integrar los avances de la genómica al derecho fundamental de la salud de todos los seres humanos. La proteómica, entendida como la disciplina científica que estudia los proteomas, es de vital importancia en la investigación en salud, ya que el conocimiento de las proteínas y moléculas efectoras de la función celular permitirá un mejor entendimiento de la fisiología humana. En este trabajo se describen los antecedentes y los conocimientos básicos del análisis proteómico basado en la espectrometría de masas y se comentan los usos de la proteómica en la búsqueda de biomarcadores para el diagnóstico y pronóstico de diferentes enfermedades, los avances en la comprensión de los trastornos crónicos y algunas enfermedades infecciosas. De manera adicional, se delinean las ventajas de la espectrometría de masas en la genotipificación de patógenos y el estudio de los polimorfismos de una sola base (SNP, por sus siglas en inglés).<br>The proteome is defined as the entirety of proteins expressed by a genome in a given time under specific physiological conditions. In an organism, the cells contain the same genome; however, they express different proteins in response to a specific micro-environment. Proteomics is responsible for the study of proteomes, using a wide range of methodological techniques. Actually, proteomics is a key tool in health research because it has made possible systematic analysis of hundreds of proteins in clinical samples with the promise of discovering new protein biomarkers for different disease conditions. Finally, proteomic strategy is a technology well-suited to provide a better understanding of systems biology and human health

Menor Resistencia Insulínica en el Obeso de Altura

It has been demonstrated that obesity produces insulin resistance, dislipidemia, high blood pressure and other metabolic alterations which are risk factors to develop atherosclerosis and cardiovascular events. On the other land, chronic expossure to high altitude increases the insulin sensibility; the blood glucose concentration, total and LDL cholesterol are lower in high altitude dwellers than in sea level inhabitants, and HDL cholesterol is higher. Therefore, it seemed interesting to study the metabolic events in the obese people living at high altitude. A total of 41 male obese, aged 20 to 60 years, were studied; 11 living at sea level (Lima, 150 m above sea level) and 30 living at high altitude (Huancayo 3200 m above sea level). Oral glucose tolerance tests consisting in 75 g were conducted in these subjects. Anthropometric measures (height, weight, body mass index, abdominal ship index) as well as blood glucose, total and HDL cholesterol, triglycerldes and insulin were measured. LDL and VLDL were calculated following the Friedewald's formula. Blood glucose, and insulin were significantly lower 2 hours after the glucose administration in the high altitude obese subjects demonstrating that obese subjects at high altitude have a lower insulin resistance, which is also supported by the higher HDL2 concentration, lower blood pressure and no hypertensive subjects. These results demonstrate that exposition to high altitude lowers the insulin resistance and the accompaning metabolic and clinical events even in obese subjects.Se ha demostrado que la obesidad produce resistencia a la insulina, dislipidemia, hipertensión arterial (HTA) y otros eventos metabólicos que favorecen la aparición de ateroesclerosis e incremento de eventos cardiovasculares. Por otro lado, la exposición a la altura incrementa la sensibilidad a la insulina, disminuye la concentración de glucosa sanguínea, colesterol total y LDL e incrementa la HDL. El presente trabajo tiene por objetivo dilucidar los cambios que se producen en el obeso expuesto a la altura. Se estudió 41 obesos, 11 de nivel del mar (Lima, 150 msnm) y 30 de altura (Huancayo- Perú, 3200 msnm), de género masculino, de 20 a 60 años de edad, a quienes se efectuó una prueba oral de tolerancia de la glucosa (75 g). Se realizaron mediciones antropométricas (talla, peso, IMC e índice cintura/cadera- ICC- ) y determinaciones en sangre de glucosa, colesterol total, HDL, triglicéridos (Tg) e insulina. El LDL y VLDL fueron calculados mediante la fórmula de Friedewald. Pese a que los obesos de altura tuvieron el IMC superior a los obesos de nivel del mar, se encontró que la glicemia e insulinemia son significativamente menores a los 120 minutos de la administración de la glucosa, indicando una menor resistencia a la insulina que, además, se sustenta en la mayor concentración de HDL2, menor PA y la ausencia de HTA. Estos resultados corroboran que la altura es un ambiente en el que disminuye la resistencia a la insulina así como los eventos clínicos y metabólicos que la acompañan, aún en sujetos obesos

Monitoring Mitochondrial Function in Aedes albopictus C6/36 Cell Line during Dengue Virus Infection

Aedes aegypti and Aedes albopictus mosquitoes are responsible for dengue virus (DENV) transmission in tropical and subtropical areas worldwide, where an estimated 3 billion people live at risk of DENV exposure. DENV-infected individuals show symptoms ranging from sub-clinical or mild to hemorrhagic fever. Infected mosquitoes do not show detectable signs of disease, even though the virus maintains a lifelong persistent infection. The interactions between viruses and host mitochondria are crucial for virus replication and pathogenicity. DENV infection in vertebrate cells modulates mitochondrial function and dynamics to facilitate viral proliferation. Here, we describe that DENV also regulates mitochondrial function and morphology in infected C6/36 mosquito cells (derived from Aedes albopictus). Our results showed that DENV infection increased ROS (reactive oxygen species) production, modulated mitochondrial transmembrane potential and induced changes in mitochondrial respiration. Furthermore, we offer the first evidence that DENV causes translocation of mitofusins to mitochondria in the C6/36 mosquito cell line. Another protein Drp-1 (Dynamin-related protein 1) did not localize to mitochondria in DENV-infected cells. This observation therefore ruled out the possibility that the abovementioned alterations in mitochondrial function are associated with mitochondrial fission. In summary, this report provides some key insights into the virus–mitochondria crosstalk in DENV infected mosquito cells

List of experimentally determined AlgU promoters of <i>P</i>. <i>aeruginosa</i> that served as a reference for generating the Position-Specific Scoring Matrix.

List of experimentally determined AlgU promoters of P. aeruginosa that served as a reference for generating the Position-Specific Scoring Matrix.</p

Total proteins identified in the <i>algU</i> mutant under vegetative growth as compared to wild-type strain.

Total proteins identified in the algU mutant under vegetative growth as compared to wild-type strain.</p

Serum metabolic alterations upon Zika infection

Zika virus (ZIKV) infection has recently emerged as a major concern worldwide due to its strong association with nervous system malformation (microcephaly) of fetuses in pregnant women infected by the virus. Signs and symptoms of ZIKV infection are often mistaken with other common viral infections. Since transmission may occur through biological fluids exchange and coitus, in addition to mosquito bite, this condition is an important infectious disease. Thus, understanding the mechanism of viral infection has become an important research focus, as well as providing potential targets for assertive clinical diagnosis and quality screening for hemoderivatives. Within this context, the present work analyzed blood plasma from 79 subjects, divided as a control group and a ZIKV-infected group. Samples underwent direct-infusion mass spectrometry and statistical analysis, where eight markers related to the pathophysiological process of ZIKV infection were elected and characterized. Among these, Angiotensin (1-7) and Angiotensin I were upregulated under infection, showing an attempt to induce autophagy of the infected cells. However, this finding is concerning about hypertensive individuals under treatment with inhibitors of the Renin-Angiotensin System (RAS), which could reduce this response against the virus and exacerbate the symptoms of the infection. Moreover, one of the most abundant glycosphingolipids in the nervous tissue, Ganglioside GM2, was also elected in the present study as an infection biomarker. Considered an important pathogen receptor at membrane's outer layer, this finding represents the importance of gangliosides for ZIKV infection and its association with brain tropism. Furthermore, a series of phosphatidylinositols were also identified as biomarkers, implying a significant role of the PI3K-AKT-mTOR Pathway in this mechanism. Finally, these pathways may also be understood as potential targets to be considered in pharmacological intervention studies on ZIKV infection management8CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPPNPD 1578388; PROEX 148974017/20614-4; 16/17066-2; 14/00302-0; 11/50400-0; 15/06809-1Plano Nacional de Enfrentamento ao Aedes aegypti e Microcefalia from the Brazilian Ministry of Healt