RESUMO
The rapid spread and public health impact of the novel SARS-CoV-2 variants that cause COVID-19 continue to produce major global impacts and social distress. Several vaccines were developed in record time to prevent and limit the spread of the infection, thus playing a pivotal role in controlling the pandemic. Although the repurposing of available drugs attempts to provide therapies of immediate access against COVID-19, there is still a need for developing specific treatments for this disease. Remdesivir, molnupiravir and Paxlovid remain the only evidence-supported antiviral drugs to treat COVID-19 patients, and only in severe cases. To contribute on the search of potential Covid-19 therapeutic agents, we targeted the viral RNA-dependent RNA polymerase (RdRp) and the exoribonuclease (ExoN) following two strategies. First, we modeled and analyzed nucleoside analogs sofosbuvir, remdesivir, favipiravir, ribavirin, and molnupiravir at three key binding sites on the RdRp-ExoN complex. Second, we curated and virtually screened a database containing 517 nucleotide analogs in the same binding sites. Finally, we characterized key interactions and pharmacophoric features presumably involved in viral replication halting at multiple sites. Our results highlight structural modifications that might lead to more potent SARS-CoV-2 inhibitors against an expansive range of variants and provide a collection of nucleotide analogs useful for screening campaigns.
RESUMO
In the past few years, our understanding of the RNA virosphere has changed dramatically due to the growth and spurt of metagenomics, exponentially increasing the number of RNA viral sequences, and providing a better understanding of their range of potential hosts. As of today, the only conserved protein among RNA viruses appears to be the monomeric RNA-dependent RNA polymerase. This enzyme belongs to the right-hand DNA-and RNA polymerases, which also includes reverse transcriptases and eukaryotic replicative DNA polymerases. The ubiquity of this protein in RNA viruses makes it a unique evolutionary marker and an appealing broad-spectrum antiviral target. In this work pairwise structural comparisons of viral RdRps and RTs were performed, including tertiary structures that have been obtained in the last few years. The resulting phylogenetic tree shows that the RdRps from (+)ss- and dsRNA viruses might have been recruited several times throughout the evolution of mobile genetic elements. RTs also display multiple evolutionary routes. We have identified a structural core comprising the entire palm, a large moiety of the fingers and the N-terminal helices of the thumb domain, comprising over 300 conserved residues, including two regions that we have named the "knuckles" and the "hypothenar eminence". The conservation of an helix bundle in the region preceding the polymerase domain confirms that (-)ss and dsRNA Reoviruses' polymerases share a recent ancestor. Finally, the inclusion of DNA polymerases into our structural analyses suggests that monomeric RNA-dependent polymerases might have diverged from B-family polymerases.
Assuntos
RNA Polimerases Dirigidas por DNA , Evolução Molecular , Sequência de Aminoácidos , DNA Polimerase Dirigida por DNA , RNA Polimerases Dirigidas por DNA/genética , Filogenia , RNA/genéticaRESUMO
Low complexity regions (LCRs) are protein sequences formed by a set of compositionally biased residues. LCRs are extremely abundant in cellular proteins and have also been reported in viruses, where they may partake in evasion of the host immune system. Analyses of 28,231 SARS-CoV-2 whole proteomes and of 261,051 spike protein sequences revealed the presence of four extremely conserved LCRs in the spike protein of several SARS-CoV-2 variants. With the exception of Iota, where it is absent, the Spike LCR-1 is present in the signal peptide of 80.57% of the Delta variant sequences, and in other variants of concern and interest. The Spike LCR-2 is highly prevalent (79.87%) in Iota. Two distinctive LCRs are present in the Delta spike protein. The Delta Spike LCR-3 is present in 99.19% of the analyzed sequences, and the Delta Spike LCR-4 in 98.3% of the same set of proteins. These two LCRs are located in the furin cleavage site and HR1 domain, respectively, and may be considered hallmark traits of the Delta variant. The presence of the medically-important point mutations P681R and D950N in these LCRs, combined with the ubiquity of these regions in the highly contagious Delta variant opens the possibility that they may play a role in its rapid spread.
Assuntos
COVID-19/genética , Mutação de Sentido Incorreto , Proteoma/genética , SARS-CoV-2/genética , Glicoproteína da Espícula de Coronavírus/genética , Substituição de Aminoácidos , COVID-19/metabolismo , Humanos , SARS-CoV-2/metabolismo , Glicoproteína da Espícula de Coronavírus/metabolismoRESUMO
As of today, there is no antiviral for the treatment of the SARS-CoV-2 infection, and the development of a vaccine might take several months or even years. The structural superposition of the hepatitis C virus polymerase bound to sofosbuvir, a nucleoside analog antiviral approved for hepatitis C virus infections, with the SARS-CoV polymerase shows that the residues that bind to the drug are present in the latter. Moreover, a multiple alignment of several SARS-CoV-2, SARS and MERS-related coronaviruses polymerases shows that these residues are conserved in all these viruses, opening the possibility to use sofosbuvir against these highly infectious pathogens.
Assuntos
Antivirais/química , Betacoronavirus/enzimologia , Infecções por Coronavirus/virologia , Pandemias/prevenção & controle , Pneumonia Viral/virologia , RNA Polimerase Dependente de RNA/química , Sofosbuvir/química , Proteínas não Estruturais Virais/química , Antivirais/uso terapêutico , Sequência de Bases , COVID-19 , Domínio Catalítico , Simulação por Computador , Infecções por Coronavirus/tratamento farmacológico , RNA-Polimerase RNA-Dependente de Coronavírus , Humanos , Coronavírus da Síndrome Respiratória do Oriente Médio/enzimologia , Pneumonia Viral/tratamento farmacológico , Ligação Proteica , Estrutura Terciária de Proteína , RNA Polimerase Dependente de RNA/genética , Coronavírus Relacionado à Síndrome Respiratória Aguda Grave/enzimologia , SARS-CoV-2 , Síndrome Respiratória Aguda Grave/tratamento farmacológico , Síndrome Respiratória Aguda Grave/virologia , Sofosbuvir/uso terapêutico , Proteínas não Estruturais Virais/genéticaRESUMO
OBJECTIVES: Yellow fever virus historically was a frequent threat to American and European coasts. Medical milestones such as the discovery of mosquitoes as vectors and subsequently an effective vaccine significantly reduced its incidence, in spite of which, thousands of cases of this deathly disease still occur regularly in Sub-Saharan Africa and the Amazonian basin in South America, which are usually not reported. An urban outbreak in Angola, consecutive years of increasing incidence near major Brazilian cities, and imported cases in China, South America and Europe, have brought this virus back to the global spotlight. The aim of this article is to underline that the preventive YFV measures, such as vaccination, need to be carefully revised in order to minimize the risks of new YFV outbreaks, especially in urban or immunologically vulnerable places. Furthermore, this article highlights the diverse factors that have favored the spread of other Aedes spp.-associated arboviral diseases like Dengue, Chikungunya and Zika, to northern latitudes causing epidemics in the United States and Europe, emphasizing the possibility that YFV might follow the path of these viruses unless enhanced surveillance and efficient control systems are urgently initiated.
Assuntos
Febre Amarela/epidemiologia , Vírus da Febre Amarela/isolamento & purificação , Animais , Humanos , Mosquitos Vetores/fisiologia , Mosquitos Vetores/virologia , América do Norte/epidemiologia , Febre Amarela/transmissão , Febre Amarela/virologia , Vírus da Febre Amarela/classificação , Vírus da Febre Amarela/genéticaRESUMO
BACKGROUND: Essential hypertension is one of the main risk factors for the development of coronary artery disease (CAD). Hypertension causes endothelial dysfunction which is considered an early sign for the development of CAD. Positron emission tomography is a non-invasive imaging technique that measures myocardial blood flow (MBF), allowing us to identify patients with endothelial dysfunction. METHODS AND RESULTS: 19 patients without comorbidities recently diagnosed hypertensive, as well as 21 healthy volunteers were studied. A three-phase (rest, cold pressor test, and adenosine-induced hyperemia) (13)N-ammonia PET was performed, and MBF was measured. Endothelial-Dependent Vasodilation Index, ΔMBF, and coronary flow reserve (CFR) were calculated for each patient. Hypertensive patients had a significantly higher systolic and diastolic blood pressures compared with the control group (134.6 ± 11.7/86.4 ± 10.6 mm Hg and 106.0 ± 11.8/71.4 ± 6.6 mm Hg, respectively, P < .001). The ENDEVI (1.28 ± 0.26 vs 1.79 ± 0.30, P < .001), the ΔMBF (0.81 ± 0.50 vs 0.25 ± 0.21, P < .001) and the CFR (2.18 ± 0.88 vs 3.17 ± 0.68, P = .001) were significantly lower in the hypertensive patients compared to the control group, 84% of the former group had endothelial dysfunction i.e., ENDEVI < 1.5 and 58% had vasomotor abnormalities, i.e., CFR < 2.5. CONCLUSIONS: In this study, we showed that recently diagnosed hypertensive patients have coronary endothelial dysfunction and vasomotor disturbances which are early signs for the development of CAD.
Assuntos
Amônia , Endotélio Vascular/fisiopatologia , Hipertensão/fisiopatologia , Radioisótopos de Nitrogênio , Tomografia por Emissão de Pósitrons/métodos , Adulto , Idoso , Estudos de Casos e Controles , Circulação Coronária , Feminino , Humanos , Hipertensão/diagnóstico por imagem , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Espécies Reativas de Oxigênio/metabolismoRESUMO
The results of a detailed bioinformatic search for ribonucleotidyl coenzyme biosynthetic sequences in DNA- and RNA viral genomes are presented. No RNA viral genome sequence available as of April 2011 appears to encode for sequences involved in coenzyme biosynthesis. In both single- and double-stranded DNA viruses a diverse array of coenzyme biosynthetic genes has been identified, but none of the viral genomes examined here encodes for a complete pathway. Although our conclusions may be constrained by the unexplored diversity of viral genomes and the biases in the construction of viral genome databases, our results do not support the possibility that RNA viruses are direct holdovers from an ancient RNA/protein world. Extrapolation of our results to evolutionary epochs prior to the emergence of DNA genomes suggest that during those early stages living entities may have depended on discontinuous genetic systems consisting of multiple small-size RNA sequences.
Assuntos
Coenzimas/biossíntese , Evolução Química , RNA/metabolismo , Vírus/enzimologia , Vírus/genética , Biologia Computacional , Genoma Viral/genética , RNA/genética , RNA Viral/genética , RNA Viral/metabolismoRESUMO
Cardiovascular imaging is one of the disciplines in cardiology with the most recent advances. This means that the teaching of Cardiology must evolve in the same way. In 2009, the American College of Cardiology published a statement, which points out that all of the cardiology residents must have basic training in every one of the cardiovascular imaging modalities available. Ischemic heart disease is the main cause of death in the world, including Mexico. Up to 43% of the patients that suffered a myocardial infarction and up to 31% of the patients with sudden cardiac death had an almost normal nuclear myocardial perfusion study in the year before the event, thus evidencing the importance of a multi-imaging approach. With the better understanding of the pathophysiological processes of coronary artery disease, new techniques have been developed that allows the detection of this disease almost from the beginning, through the detection of endothelial dysfunction by Positron Emission Tomography. Later on, when the patient develops diffuse atherosclerosis, we can rely on the use of de coronary calcium score and the detection of atherosclerotic plaques with coronary computed tomography angiography. To detect the presence of myocardial ischemia, two methods are widely used: echocardiography and nuclear medicine. Other options to identify myocardial ischemia are magnetic resonance imaging and computed tomography, due to the development of the "Dual Source" and "Flash" technologies. After an acute coronary event, cardiovascular imaging is useful for risk stratification and detection of myocardial viability, being the positron emission tomography the gold standard.
Assuntos
Técnicas de Imagem Cardíaca , Cardiopatias/diagnóstico , Imagem Multimodal , HumanosRESUMO
La imagen cardiovascular es una de las disciplinas que más ha evolucionado en el campo de la cardiología. Ante esto, la enseñanza de la cardiología debe moverse a la par. En 2009, el Colegio Americano de Cardiología decidió publicar una declaración en la que señala que: todos los residentes de cardiología deben llevar un entrenamiento básico en cada una de las técnicas de imagen cardiovascular disponibles. La cardiopatía isquémica es la principal causa de muerte en casi todo el mundo, incluido México. Hasta 43% de los pacientes que habían sufrido un infarto del miocardio y 31% de los pacientes con muerte súbita de origen cardiaco, tenían un estudio de perfusión por medicina nuclear prácticamente normal en el año previo al desenlace, poniendo en evidencia la importancia del abordaje por medio de distintos métodos de imagen. Con el mejor entendimiento de los procesos fisiopatológicos de la enfermedad arterial coronaria, se han desarrollado técnicas diagnósticas que nos permiten identificar esta patología prácticamente desde su inicio, a través de la detección de disfunción endotelial por medio de la tomografía por emisión de positrones. Más adelante, cuando los pacientes desarrollan ateroesclerosis manifiesta, contamos con herramientas como el score de calcio y la detección de las placas ateroscleróticas por medio de la tomografía computarizada. Para detectar la presencia de isquemia miocárdica contamos con dos métodos ampliamente utilizados: la ecocardiografía en estrés con dobutamina o dipiridamol y la medicina nuclear. Otras opciones para la identificación de isquemia son la resonancia magnética y la tomografía computada, gracias a la tecnología Dual Source y Flash. Posterior a un evento coronario, la imagen cardiovascular tiene como funciones la estratificación de riesgo y la detección de tejido miocárdico viable, siendo hoy en día el método de elección la tomografía por emisión de positrones.
Cardiovascular imaging is one of the disciplines in cardiology with the most recent advances. This means that the teaching of Cardiology must evolve in the same way. In 2009, the American College of Cardiology published a statement, which points out that all of the cardiology residents must have basic training in every one of the cardiovascular imaging modalities available. Ischemic heart disease is the main cause of death in the world, including Mexico. Up to 43% of the patients that suffered a myocardial infarction and up to 31% of the patients with sudden cardiac death had an almost normal nuclear myocardial perfusion study in the year before the event, thus evidencing the importance of a multi-imaging approach. With the better understanding of the pathophysiological processes of coronary artery disease, new techniques have been developed that allows the detection of this disease almost from the beginning, through the detection of endothelial dysfunction by Positron Emission Tomography. Later on, when the patient develops diffuse atherosclerosis, we can rely on the use of de coronary calcium score and the detection of atherosclerotic plaques with coronary computed tomography angiography. To detect the presence of myocardial ischemia, two methods are widely used: echocardiography and nuclear medicine. Other options to identify myocardial ischemia are magnetic resonance imaging and computed tomography, due to the development of the "Dual Source" and "Flash" technologies. After an acute coronary event, cardiovascular imaging is useful for risk stratification and detection of myocardial viability, being the positron emission tomography the gold standard.
Assuntos
Humanos , Técnicas de Imagem Cardíaca , Cardiopatias/diagnóstico , Imagem MultimodalRESUMO
Objetivos: Determinar la aplicación que tiene la tomografía por emisión de positrones en el seguimiento de pacientes con arteritis de Takayasu con actividad inflamatoria y su correlación con los criterios clínicos establecidos. Métodos: Se incluyeron 35 pacientes con diagnóstico de arteritis de Takayasu. Se determinó velocidad de sedimentación globular, proteína C reactiva, biometría hemática, así como, fibrinógeno y se aplicaron los criterios clínicos de actividad. Se realizó tomografía por emisión de positrones basal de los pacientes positivos para actividad inflamatoria, todos recibieron tratamiento farmacológico. De forma aleatoria se incluyó a 10 pacientes que posterior al tratamiento durante seis meses se les realizó un nuevo estudio clínico y una tomografía por emisión de positrones para determinar actividad inflamatoria. Se compararon los criterios clínicos con tomografía por emisión de positrones tanto del estudio basal como el de seguimiento. Resultados: Los criterios clínicos tuvieron una sensibilidad de 63% y especificidad de 90% para demostrar actividad inflamatoria en forma basal. La sensibilidad de los criterios clínicos disminuyó posterior al tratamiento hasta 27%, en donde se observó que pacientes aparentemente inactivos por clínica, continuaban activos por tomografía por emisión de positrones. Discusión: Éste es el primer estudio que compara de manera prospectiva los hallazgos de tomografía por emisión de positrones antes y después del tratamiento para actividad inflamatoria en pacientes con arteritis de Takayasu. Los criterios clínicos carecen de sensibilidad para la detección de actividad inflamatoria en el seguimiento posterior al tratamiento. Conclusiones: El tomografía por emisión de positrones es una técnica de diagnóstico con una alta sensibilidad y especificidad para el diagnóstico y seguimiento de pacientes con arteritis de Takayasu y actividad inflamatoria.
Objective: To determine the application of PET in monitoring patients with Takayasu's arteritis (TA) with inflammatory activity (IA) and its correlation with established clinical criteria. Methods: 35 patients diagnosed with TA were enrolled. Determination of erythrocyte sedimentation rate, C-reactive protein, fibrinogen and a complete blood count was performed and clinical criteria of activity were applied. A baseline PET was performed in all patients. Those who were positive for IA received drug treatment. Among the group of active patients, ten were randomized to undergo another PET scan and clinical criteria determination to evaluate inflammatory activity after 6 months of treatment. We compared clinical criteria with PET capacity to determine IA. The results between the initial and final PET were finally compared. Results: Clinical criteria had a sensitivity of 63% and a specificity of 90% to show IA. Sensitivity decreased after 6 months of treatment to 27%. Discussion: This is the first prospective study that compares the findings of PET before and after treatment for IA in patients with TA. Clinical criteria lack sensitivity for the detection of IA in the follow-up after treatment. Conclusions: PET is a diagnostic technique with high sensitivity and specificity for the diagnosis and monitoring of patients with TA and inflammatory activity.
Assuntos
Adulto , Feminino , Humanos , Masculino , Tomografia por Emissão de Pósitrons , Arterite de Takayasu , Algoritmos , Inflamação , Estudos Longitudinais , Estudos ProspectivosRESUMO
BACKGROUND: Dyslipidemias constitute an independent risk factor for the development of atherogenesis and they also predispose to the development of endothelial dysfunction (ED). Using PET with (13)N-ammonia, it is possible to quantify myocardial blood flow (MBF) in mL/min/g and to quantitatively evaluate ED. With the use of lipid lowering therapy it is possible to reduce ED and increase the MBF and the endothelial-dependent vasodilation index (ENDEVI). In this study, we aimed to evaluate with (13)N-ammonia PET the benefic effects of the combined treatment ezetimibe/simvastatine on the endothelial function of dyslipidemic patients after 8 weeks of treatment. MATERIAL AND METHODS: Fourteen consecutive patients with dyslipidemia diagnosis and 17 healthy volunteers were studied with a three phase [rest, Cold Pressor Test (CPT), and adenosine-induced hyperemia] (13)N-ammonia PET for MBF quantification assessment. A second PET study was performed in the dyslipidemic group after 8 weeks of treatment with ezetimibe/simvastatine (10/40 mg). Myocardial flow reserve (MFR), ENDEVI, and %ΔMBF were calculated. RESULTS: Total cholesterol, LDL cholesterol, HDL cholesterol, and triglycerides concentrations were markedly altered in the dyslipidemic group and after 8 weeks of treatment these values improved. Dyslipidemic patients showed endothelial dysfunction when compared with the control group, (MFR 2.79 ± 0.94 vs 3.15 ± 0.48, P < 0.05 ; ENDEVI 1.28 ± 0.25 vs 1.53 ± 0.24, P < 0.05; and %ΔMBF 29.08 ± 24.62 vs 53 ± 24.60%, P < 0.05, respectively). After 8 weeks of treatment, we found a significant increase in all the endothelial function markers (MFR: 3.14 ± 0.86, P < 0.05, ENDEVI 1.65 ± 0.23, P < 0.05; %ΔMBF: 65.21 ± 23.43, P < 0.05). CONCLUSIONS: Dyslipidemic patients show endothelial dysfunction measured with (13)N-ammonia PET. Treatment with ezetimibe/simvastatine was effective improving the lipid profile as well as the endothelial function of these patients. PET may be a useful tool to monitor vascular reactivity and regression/progression of coronary atherosclerosis after pharmacologic interventions.
Assuntos
Amônia/química , Azetidinas/farmacologia , Dislipidemias/tratamento farmacológico , Endotélio Vascular/patologia , Isótopos de Nitrogênio/farmacologia , Tomografia por Emissão de Pósitrons/métodos , Sinvastatina/farmacologia , Adenosina/metabolismo , Adolescente , Adulto , Idoso , Anticolesterolemiantes/farmacologia , Estudos de Casos e Controles , Dislipidemias/diagnóstico , Ezetimiba , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Miocárdio/patologia , Estudos Prospectivos , Fatores de RiscoRESUMO
BACKGROUND AND AIMS: We undertook this study to evaluate the functional impact of coronary abnormalities in patients with suspected coronary artery disease (CAD) by means of integrated positron emission tomography (PET) and coronary computed tomography angiography (CCTA) scan obtained on a hybrid state-of-the-art PET/CT scanner. METHODS: We studied 29 consecutive, patients with a clinically suspected intermediate risk for CAD, using a hybrid PET/CT 64 slice scanner. During a single scanning session, CCTA was performed for coronary anatomy evaluation, and a rest/adenosine stress (13)N-ammonia PET was performed for myocardial perfusion assessment in 3D mode with CT attenuation correction. RESULTS: Twenty four (82.7%) patients had atherosclerosis detected by CCTA; 15 patients had significant (≥50%) coronary stenoses and all 15 patients showed ischemia by PET; moreover, 10/15 patients had a Summed Stress Score >12.20/24 and 83.3% patients with atherosclerosis detected by CCTA showed ischemia by PET. Two of five patients with normal coronary arteries showed ischemia by PET. CCTA agreement in positive identification of PET ischemia was 91% and agreement in ruling out ischemia was 43%; PET agreement in detecting CCTA atherosclerosis was 83%, and agreement in ruling it out was 60%. CONCLUSIONS: We found a strong relation between significant coronary stenosis identified by CCTA and ischemia by PET. However, in cases with low-grade stenosis, PET scan can assess the functional significance of atherosclerotic abnormalities.